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CHEMISTRY (638 journals)                  1 2 3 4 | Last

Showing 1 - 200 of 735 Journals sorted alphabetically
2D Materials     Hybrid Journal   (Followers: 14)
Accreditation and Quality Assurance: Journal for Quality, Comparability and Reliability in Chemical Measurement     Hybrid Journal   (Followers: 29)
ACS Catalysis     Hybrid Journal   (Followers: 49)
ACS Chemical Neuroscience     Hybrid Journal   (Followers: 23)
ACS Combinatorial Science     Hybrid Journal   (Followers: 23)
ACS Macro Letters     Hybrid Journal   (Followers: 27)
ACS Medicinal Chemistry Letters     Hybrid Journal   (Followers: 42)
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Acta Chemica Iasi     Open Access   (Followers: 6)
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Acta Chromatographica     Full-text available via subscription   (Followers: 8)
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adhäsion KLEBEN & DICHTEN     Hybrid Journal   (Followers: 8)
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Adsorption Science & Technology     Open Access   (Followers: 7)
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African Journal of Bacteriology Research     Open Access  
African Journal of Chemical Education     Open Access   (Followers: 4)
African Journal of Pure and Applied Chemistry     Open Access   (Followers: 8)
Agrokémia és Talajtan     Full-text available via subscription   (Followers: 2)
Al-Kimia : Jurnal Penelitian Sains Kimia     Open Access  
Alchemy : Journal of Chemistry     Open Access   (Followers: 3)
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American Journal of Biochemistry and Molecular Biology     Open Access   (Followers: 22)
American Journal of Chemistry     Open Access   (Followers: 32)
American Journal of Plant Physiology     Open Access   (Followers: 13)
American Mineralogist     Hybrid Journal   (Followers: 15)
Anadolu University Journal of Science and Technology A : Applied Sciences and Engineering     Open Access  
Analyst     Full-text available via subscription   (Followers: 37)
Angewandte Chemie     Hybrid Journal   (Followers: 185)
Angewandte Chemie International Edition     Hybrid Journal   (Followers: 268)
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Annual Review of Chemical and Biomolecular Engineering     Full-text available via subscription   (Followers: 12)
Annual Review of Food Science and Technology     Full-text available via subscription   (Followers: 14)
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Arabian Journal of Chemistry     Open Access   (Followers: 6)
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Atomization and Sprays     Full-text available via subscription   (Followers: 4)
Australian Journal of Chemistry     Hybrid Journal   (Followers: 7)
Autophagy     Hybrid Journal   (Followers: 3)
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Biochemical Pharmacology     Hybrid Journal   (Followers: 11)
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Biomedical Chromatography     Hybrid Journal   (Followers: 6)
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BioNanoScience     Partially Free   (Followers: 5)
Bioorganic & Medicinal Chemistry     Hybrid Journal   (Followers: 137)
Bioorganic & Medicinal Chemistry Letters     Hybrid Journal   (Followers: 93)
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Biosensors     Open Access   (Followers: 2)
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Bitácora Digital     Open Access  
Boletin de la Sociedad Chilena de Quimica     Open Access  
Bulletin of the Chemical Society of Ethiopia     Open Access   (Followers: 1)
Bulletin of the Chemical Society of Japan     Full-text available via subscription   (Followers: 25)
Bulletin of the Korean Chemical Society     Hybrid Journal   (Followers: 1)
C - Journal of Carbon Research     Open Access   (Followers: 3)
Cakra Kimia (Indonesian E-Journal of Applied Chemistry)     Open Access  
Canadian Association of Radiologists Journal     Full-text available via subscription   (Followers: 2)
Canadian Journal of Chemistry     Hybrid Journal   (Followers: 11)
Canadian Mineralogist     Full-text available via subscription   (Followers: 6)
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Catalysis for Sustainable Energy     Open Access   (Followers: 8)
Catalysis Reviews: Science and Engineering     Hybrid Journal   (Followers: 9)
Catalysis Science and Technology     Hybrid Journal   (Followers: 9)
Catalysis Surveys from Asia     Hybrid Journal   (Followers: 3)
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Chemical and Engineering News     Free   (Followers: 22)
Chemical Bulletin of Kazakh National University     Open Access  
Chemical Communications     Full-text available via subscription   (Followers: 75)
Chemical Engineering Research and Design     Hybrid Journal   (Followers: 27)
Chemical Research in Chinese Universities     Hybrid Journal   (Followers: 3)
Chemical Research in Toxicology     Hybrid Journal   (Followers: 22)
Chemical Reviews     Hybrid Journal   (Followers: 216)
Chemical Science     Open Access   (Followers: 28)
Chemical Technology     Open Access   (Followers: 35)
Chemical Vapor Deposition     Hybrid Journal   (Followers: 5)
Chemie in Unserer Zeit     Hybrid Journal   (Followers: 58)
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Chemistry     Open Access  
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Chemistry & Biology     Full-text available via subscription   (Followers: 33)
Chemistry & Industry     Hybrid Journal   (Followers: 8)
Chemistry - A European Journal     Hybrid Journal   (Followers: 176)
Chemistry - An Asian Journal     Hybrid Journal   (Followers: 16)
Chemistry and Materials Research     Open Access   (Followers: 21)
Chemistry Central Journal     Open Access   (Followers: 4)
Chemistry Education Research and Practice     Free   (Followers: 5)
Chemistry in Education     Open Access   (Followers: 9)
Chemistry International     Open Access   (Followers: 3)
Chemistry Letters     Full-text available via subscription   (Followers: 46)
Chemistry of Materials     Hybrid Journal   (Followers: 282)
Chemistry of Natural Compounds     Hybrid Journal   (Followers: 10)
Chemistry World     Full-text available via subscription   (Followers: 21)
Chemistry-Didactics-Ecology-Metrology     Open Access   (Followers: 1)
ChemistryOpen     Open Access   (Followers: 1)
Chemkon - Chemie Konkret, Forum Fuer Unterricht Und Didaktik     Hybrid Journal  
Chemoecology     Hybrid Journal   (Followers: 3)
Chemometrics and Intelligent Laboratory Systems     Hybrid Journal   (Followers: 15)
Chemosensors     Open Access  
ChemPhysChem     Hybrid Journal   (Followers: 12)
ChemPlusChem     Hybrid Journal   (Followers: 2)
ChemTexts     Hybrid Journal  
CHIMIA International Journal for Chemistry     Full-text available via subscription   (Followers: 2)
Chinese Journal of Chemistry     Hybrid Journal   (Followers: 6)
Chinese Journal of Polymer Science     Hybrid Journal   (Followers: 11)
Chromatographia     Hybrid Journal   (Followers: 22)
Chromatography     Open Access   (Followers: 3)
Chromatography Research International     Open Access   (Followers: 5)
Cogent Chemistry     Open Access   (Followers: 2)
Colloid and Interface Science Communications     Open Access  
Colloid and Polymer Science     Hybrid Journal   (Followers: 11)
Colloids and Interfaces     Open Access  
Colloids and Surfaces B: Biointerfaces     Hybrid Journal   (Followers: 7)
Combinatorial Chemistry & High Throughput Screening     Hybrid Journal   (Followers: 4)
Combustion Science and Technology     Hybrid Journal   (Followers: 23)
Comments on Inorganic Chemistry: A Journal of Critical Discussion of the Current Literature     Hybrid Journal   (Followers: 2)
Communications Chemistry     Open Access   (Followers: 1)
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Computational Chemistry     Open Access   (Followers: 2)
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Coordination Chemistry Reviews     Full-text available via subscription   (Followers: 4)
Copernican Letters     Open Access   (Followers: 1)
Corrosion Series     Full-text available via subscription   (Followers: 7)
Critical Reviews in Biochemistry and Molecular Biology     Hybrid Journal   (Followers: 8)
Croatica Chemica Acta     Open Access  
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Current Science     Open Access   (Followers: 75)
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Dalton Transactions     Full-text available via subscription   (Followers: 26)
Detection     Open Access   (Followers: 4)

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Journal Cover
Biochemical Pharmacology
Journal Prestige (SJR): 1.832
Citation Impact (citeScore): 5
Number of Followers: 11  
  Hybrid Journal Hybrid journal (It can contain Open Access articles)
ISSN (Print) 0006-2952
Published by Elsevier Homepage  [3160 journals]
  • Treatment of polymyalgia rheumatica
    • Abstract: Publication date: Available online 20 March 2019Source: Biochemical PharmacologyAuthor(s): Santos Castañeda, Noelia García-Castañeda, Diana Prieto-Peña, Dolores Martínez-Quintanilla, Esther F. Vicente, Ricardo Blanco, Miguel A. González-Gay Polymyalgia rheumatica (PMR) is an inflammatory disease characterized by bilateral pain involving predominantly the shoulders and proximal aspects of the arms and less commonly the neck and the pelvic girdle. This review discusses briefly the main epidemiological data and clinical features of this condition. Especial attention is paid in the management of the disease. For this reason, both the classic management and the impact of new therapies are discussed in depth. In general, patients with PMR experience a rapid response to 12.5-25 mg/prednisone/day in less than a week. Patients with poor response to glucocorticoids or with relapsing disease require other therapies aimed mainly to spare glucocorticoids. Among them, methotrexate is the most commonly used. Nevertheless, different studies indicate that this agent yields only a modest effect. Biologic therapies against the main cytokines involved in the pathogenesis of the disease have been used in refractory patients. However, randomized controlled trials do not support the use of anti-tumor necrosis factor agents in PMR. In contrast, several case series and retrospective studies highlight the efficacy of the anti-interleukin-6 receptor tocilizumab in PMR. Nonetheless, controlled trials are needed to fully establish the beneficial effect of this agent. The potential favorable effect of the Janus-kinase inhibitors and new anti-interleukin-6 antagonists remains to be determined.Graphical abstractRelationship of PMR with GCA and other related inflammatory diseases in the elderly. DMARDs: disease-modifying anti-rheumatic drugs; EORA: elderly-onset rheumatoid arthritis; IL: interleukins (1, 6, 8); MCP-1: monocyte chemoattractant protein 1; NSAIDs: non-steroidal anti-inflammatory drugs; PMR: polymyalgia rheumatica; RS3PE: remitting seronegative symmetrical synovitis with pitting oedema syndrome; TNF-α: tumor necrosis factor alpha; Th: T-helper (Th1, Th17) lymphocytes; Treg: regulatory T lymphocytes. The two images at the center bottom show a subacromial bursitis and tenosynovitis of the biceps in a patient with PMR. The one on the right at the middle exemplifies the “halo sign” in a temporal artery of a patient with giant cell arteritis.Graphical abstract for this article
  • Targeting NFE2L2, a transcription factor upstream of MMP-2: A potential
           therapeutic strategy for temozolomide resistant glioblastoma
    • Abstract: Publication date: Available online 15 March 2019Source: Biochemical PharmacologyAuthor(s): Y. Rajesh, Angana Biswas, Utkarsh Kumar, Payel Banik, Rashmi Bharti, Santoshi Nayak, Sudip K. Ghosh, Mahitosh Mandal Glioblastoma (GBM) is the most malignant form of brain tumor posing a major threat to cancer amelioration. Temozolomide (TMZ) resistance is one of the major hurdles towards GBM prognosis. Oxidative stress and ECM remodeling are the two important processes involved in gaining chemo-resistance. Here, we established NFE2L2, an important member of oxidative stress regulation elevated in resistant cells, to be playing a transcriptional regulatory role on MMP-2, an ECM remodeling marker. This link led us to further explore targeted molecules to inhibit NFE2L2, thus affecting MMP-2, an important member promoting chemo-resistance. Thus, diosgenin was proposed as a novel NFE2L2 inhibitor acting as an alternative strategy to prevent the high dose administration of TMZ. Combinatorial therapy of diosgenin and TMZ significantly reduced the dosage regimen of TMZ and also showed affectivity in hitherto TMZ resistant GBM cells. GBM cells underwent apoptosis and early cell cycle arrest with significant reduction in MMP-2 levels. Thus preclinical validation of molecular interaction between diosgenin and NFE2L2 down-regulating MMP-2, EMT markers and promoting apoptosis, offers rationale for new therapeutic horizons in the field of glioblastoma management.Graphical abstractGraphical abstract for this article
  • High-selective HDAC6 inhibitor promotes HDAC6 degradation following
           autophagy modulation and enhanced antitumor immunity in glioblastoma
    • Abstract: Publication date: Available online 15 March 2019Source: Biochemical PharmacologyAuthor(s): Jia-Rong Liu, Chao-Wu Yu, Pei-Yun Hung, Ling-Wei Hsin, Ji-Wang Chern Glioblastoma is the most fatal type of primary brain cancer, and current treatments for glioblastoma are insufficient. HDAC6 is overexpressed in glioblastoma, and siRNA-mediated knockdown of HDAC6 inhibits glioma cell proliferation. Herein, we report a high-selective HDAC6 inhibitor, J22352, which has PROTAC (proteolysis-targeting chimeras)-like property resulted in both p62 accumulation and proteasomal degradation, leading to proteolysis of aberrantly overexpressed HDAC6 in glioblastoma. The consequences of decreased HDAC6 expression in response to J22352 decreased cell migration, increased autophagic cancer cell death and significant tumor growth inhibition. Notably, J22352 reduced the immunosuppressive activity of PD-L1, leading to the restoration of host anti-tumor activity. These results demonstrate that J22352 promotes HDAC6 degradation and induces anticancer effects by inhibiting autophagy and eliciting the antitumor immune response in glioblastoma. Therefore, this highly selective HDAC6 inhibitor can be considered a potential therapeutic for the treatment of glioblastoma and other cancers.Graphical abstractGraphical abstract for this article
  • Revisiting the combinatorial potential of cytokine subunits in the IL-12
    • Abstract: Publication date: Available online 15 March 2019Source: Biochemical PharmacologyAuthor(s): Sammy Detry, Katarzyna Składanowska, Marnik Vuylsteke, Savvas N. Savvides, Yehudi Bloch The four core members of the Interleukin-12 (IL-12) family of cytokines, IL-12, IL-23, IL-27 and IL-35 are heterodimers which share α- and β-cytokine subunits. All four cytokines are immune modulators and have been proposed to play divergent roles in inflammatory arthritis. In recent years additional combinations of α- and β-cytokine subunits belonging to the IL-12 family have been proposed to form novel cytokines such as IL-39. However, the actual extent of the combinatorial potential of the cytokine subunits in the human IL-12 family is not known. Here, we identify several combinations of subunits that form secreted heterodimeric assemblies based on a systematic orthogonal approach. The heterodimers are detected in the conditioned media harvested from mammalian cell cultures transfected with unfused pairs of cytokine subunits. While certain previously reported subunit combinations could not be recapitulated, our approach showed robustly that all four of the canonical members could be secreted. Furthermore, we provide evidence for the interaction between Cytokine Receptor Like Factor 1 (CRLF1) and Interleukin-12 subunit alpha (p35). Similar to IL-27 and IL-35 this novel heterodimer is not abundantly secreted rendering isolation from the conditioned medium very challenging, unlike IL-12 and IL-23. Our findings set the stage for fine-tuning approaches towards the biochemical reconstitution of IL-12 family cytokines for biochemical, cellular, and structural studies.Graphical abstractGraphical abstract for this article
  • The osteogenic effect of liraglutide involves enhanced mitochondrial
           biogenesis in osteoblasts
    • Abstract: Publication date: Available online 15 March 2019Source: Biochemical PharmacologyAuthor(s): Subhashis Pal, Shailendra K. Maurya, Sourav Chattopadhyay, Shyamsundar PalChina, Konica Porwal, Chirag Kulkarni, Sabyasachi Sanyal, Rohit A. Sinha, Naibedya Chattopadhyay Liraglutide (Lira), a long-acting glucagon-like peptide 1 receptor (GLP-1R) agonist reduces glycosylated hemoglobin in type 2 diabetes mellitus patients. Lira is reported to have bone conserving effect in ovariectomized (OVX) rats. Here, we investigated the osteoanabolic effect of Lira and studied the underlying mechanism. In established osteopenic OVX rats, Lira completely restored bone mass and strength comparable to parathyroid hormone (PTH 1-34). Body mass index normalized bone mineral density of Lira was higher than PTH. The serum levels of osteogenic surrogate pro-collagen type 1 N-terminal pro-peptide (P1NP) and surface referent bone formation parameters were comparable between Lira and PTH. GLP-1R, adiponectin receptor 1 (AdipoR1) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) levels in bones were downregulated in the OVX group but restored in the Lira group whereas PTH had no effect. In cultured osteoblasts, Lira time-dependently increased GLP-1R, AdipoR1 and PGC-1 α expression. In osteoblasts, Lira rapidly phosphorylated AMP-dependent protein kinase (AMPK), the cellular energy sensor. Exendin 3, a selective GLP-1R antagonist and PKA inhibitor H89 blocked Lira-induced increases in osteoblast differentiation, and expression levels of AdipoR1 and PGC-1 α. Furthermore, H89 inhibited Lira-induced phosphorylation of AMPK and dorsomorphin, an AMPK inhibitor blocked the Lira-induced increases in osteoblast differentiation and AdipoR1 and PGC-1 α levels. Lira increased mitochondrial number, respiratory proteins and respiration in osteoblasts in vitro and in vivo, and blocking mitochondrial respiration mitigated Lira-induced osteoblast differentiation. Taken together, our data show that Lira has a strong osteoanabolic effect which involves upregulation of mitochondrial function.Graphical abstractGraphical abstract for this article
  • Modulation of innate immunity by Cyclosporine A
    • Abstract: Publication date: Available online 15 March 2019Source: Biochemical PharmacologyAuthor(s): Alex M. Liddicoat, Ed C. Lavelle Cyclosporine A has long been known to suppress T cell responses by inhibiting the production of IL-2, which drives T cell proliferation, enabling its use as a therapeutic for transplantation or autoimmunity. However, cyclosporine A also impacts on innate immune cells including dendritic cells, macrophages and neutrophils. In dendritic cells, which are essential for T cell priming, cyclosporine A can modulate both expression of surface molecules than engage with T cells and cytokine secretion, leading to altered induction of T cell responses. In macrophages and neutrophils, which play key antimicrobial roles, cyclosporine A reduces the production of cytokines that can play protective roles against pathogens. Some of these molecules, if produced in the context of chronic disease, can also contribute to pathology. There have been a number of elegant recent studies addressing the mechanisms by which cyclosporine A can modulate innate immunity. In particular, cyclosporine A inhibits the release of mitochondrial factors that stimulate the production of type 1 interferons by innate immune cells. This review addresses the emerging literature on modulation of innate immune responses by cyclosporine A, its resultant impact on adaptive immune responses and how this offers potential for new therapeutic applications.Graphical abstractGraphical abstract for this article
  • Non-NAD-like PARP-1 inhibitors in prostate cancer treatment
    • Abstract: Publication date: Available online 15 March 2019Source: Biochemical PharmacologyAuthor(s): Yaroslava Karpova, Chao Wu, Ali Divan, Mark E. McDonnell, Elizabeth Hewlett, Peter Makhov, John Gordon, Min Ye, Allen Reitz, Wayne E. Childers, Tomasz Skorski, Vladimir Kolenko, Alexei V. Tulin In our previous studies of the molecular mechanisms of poly(ADP-ribose) polymerase 1 (PARP-1)-mediated transcriptional regulation we identified a novel class of PARP-1 inhibitors targeting the histone-dependent route of PARP-1 activation. Because histone-dependent activation is unique to PARP-1, non-NAD-like PARP-1 inhibitors have the potential to bypass the off-target effects of classical NAD-dependent PARP-1 inhibitors, such as olaparib, veliparib, and rucaparib. Furthermore, our recently published studies demonstrate that, compared to NAD-like PARP-1 inhibitors that are used clinically, the non-NAD-like PARP-1 inhibitor 5F02 exhibited superior antitumor activity in cell and animal models of human prostate cancer (PC). In this study, we further evaluated the antitumor activity of 5F02 and several of its novel analogues against PC cells. In contrast to NAD-like PARP-1 inhibitors, non-NAD-like PARP-1 inhibitors demonstrated efficacy against androgen-dependent and -independent routes of AR signaling activation. Our experiments reveal that methylation of the quaternary ammonium salt and the presence of esters were critical for the antitumor activity of 5F02 against PC cells. In addition, we examined the role of a related regulatory protein of PARP-1, called Poly(ADP-ribose) glycohydrolase (PARG), in prostate carcinogenesis. Our study reveals that PARG expression is severely disrupted in PC cells, which is associated with decreased integrity and localization of Cajal bodies (CB). Overall, the results of our study strengthen the justification for using non-NAD-like PARP-1 inhibitors as a novel therapeutic strategy for the treatment of advanced prostate cancer.Graphical abstractTherapeutic approaches targeting the androgen receptor (AR) signaling axis. PARP-1 inhibitors suppress AR transcriptional function and can therefore be effective against both androgen-dependent and -independent mechanisms of AR activation.Graphical abstract for this article
  • Fighting rheumatoid arthritis: Kv1.3 as a therapeutic target
    • Abstract: Publication date: Available online 14 March 2019Source: Biochemical PharmacologyAuthor(s): Antonio Serrano-Albarrás, Sergi Cirera-Rocosa, Daniel Sastre, Irene Estadella, Antonio Felipe Rheumatoid arthritis (RA) is a serious autoimmune disease that has severe impacts on both the wellbeing of patients and the economy of the health system. Similar to many autoimmune diseases, RA concurs with a long evolution, which eventually results in highly debilitating symptoms. Therapeutic treatments last for long periods during RA. However, their efficiency and side effects result in suboptimal conditions. Therefore, the need for specific, safer and nontoxic alternatives for the treatment of RA is essential.Kv1.3 is a voltage-gated potassium channel that has a crucial role in immune system response. The proliferation and activation of leukocytes are linked to differential expressions of this channel. The evidence is particularly relevant in the aggressive T effector memory (TEM) cells, which are the main actors in the development of autoimmune diseases. Blockage of Kv1.3 inhibits the reactivity of these cells. Furthermore, pharmacological inhibition of Kv1.3 ameliorates symptoms in animal models of autoimmune diseases, such as experimental autoimmune encephalomyelitis or induced psoriasis with no side effects. Kv1.3 is sensitive to several animal toxins and plant compounds, and several research groups have searched for new Kv1.3 blockers by improving these natural molecules. The research is mainly focused on enhancing the selectivity of the blockers, thereby reducing the potential for side effects on other related channel subunits. Higher selectivity means that treatments will potentially be less harmful. This leads to a lower discontinuation rate of the therapy than the current first-line treatment for RA. The molecular backgrounds of many autoimmune diseases implicate leukocyte Kv1.3 and suggests that a new medication for RA is feasible. Therapies could also be later repurposed to treat other immune system disorders.Graphical abstractGraphical abstract for this article
  • Useful message in choosing optimal biological agents for patients with
           autoimmune arthritis
    • Abstract: Publication date: Available online 13 March 2019Source: Biochemical PharmacologyAuthor(s): Jenn-Haung Lai, Xiao Chun Ling, Ling-Jun Ho The introduction of biological disease-modifying antirheumatic drug (bDMARD) treatments for various types of autoimmune arthritis, such as rheumatoid arthritis, psoriatic arthropathy and ankylosing spondylitis, represents a new era of treatment for patients with a refractory response to conventional synthetic DMARDs (csDMARDs). Many new bDMARDs with different modalities or that target different pro-inflammatory molecules, likely cytokines, are rapidly emerging. Hence, physicians in the field may be confused about choosing appropriate bDMARDs for their patients. Considering the high cost of bDMARDs and the rapid destructive process of autoimmune arthritis in patients, the choice of optimal bDMARDs for patients who fail to respond or show an inadequate therapeutic response to csDMARDs designed to control the disease is very critical. Here, we summarize the strengths and weaknesses of bDMARDs and specifically focus on their uses in patients with comorbid conditions or with specific medical conditions, such as pregnancy. This commentary provides a solid up-to-date review on commercially available bDMARDs and very useful information for physicians to facilitate the choice of more appropriate bDMARDs to treat patients with autoimmune arthritis and for basic researchers to understand the current strategies of bDMARD usage and hopefully to develop more powerful bDMARDs with fewer safety concerns.Graphical abstractGraphical abstract for this article
  • Late autophagy inhibitor chloroquine improves efficacy of the histone
           deacetylase inhibitor SAHA and temozolomide in gliomas
    • Abstract: Publication date: Available online 13 March 2019Source: Biochemical PharmacologyAuthor(s): Rosângela Mayer Gonçalves, Jonathan Paulo Agnes, Marina Delgobo, Priscila Oliveira de Souza, Marcos P. Thomé, Luana Heimfarth, Guido Lenz, José Cláudio Fonseca Moreira, Alfeu Zanotto-Filho Glioblastoma multiforme is the most aggressive type of primary brain tumor associated with few therapeutic opportunities and poor prognosis. In this study, we evaluated the efficacy of combining temozolomide (TMZ) with suberoylanilide hydroxamic acid (SAHA) - a specific histone deacetylases inhibitor – in glioma models in vitro and in vivo. In glioma cell lines, combined TMZ/SAHA promoted more cytotoxicity, G2/M arrest and apoptosis than either drugs alone. G2/M arrest was detected as soon as 24 h post drug exposure and preceded apoptosis, which occurred from 72 h treatment. TMZ and SAHA, alone or combined, also stimulated autophagy as evaluated by means of acridine orange staining and immunodetection of LC3I-II conversion and p62/SQSTM1 degradation. Time-course of autophagy accompanied G2/M arrest and preceded apoptosis, and blockage of late steps of autophagy with chloroquine (CQ) augmented SAHA/TMZ toxicity leading to apoptosis. In orthotopic gliomas in vivo, combined SAHA/TMZ showed better antitumor efficacy than either drugs alone, and adding CQ to the regimen improved antiglioma effects of SAHA and TMZ monotherapies without further benefit on combined SAHA/TMZ. In summary, the herein presented data suggest that autophagy acts as a protective response that impairs efficacy of SAHA and TMZ. Inhibiting autophagy termination with CQ may offer means to improve antitumor effects of SAHA and TMZ in gliomas and possibly other cancers.Graphical abstractGraphical abstract for this article
  • Regulation of fibroblast-like synoviocyte transformation by transcription
           factors in arthritic diseases
    • Abstract: Publication date: Available online 13 March 2019Source: Biochemical PharmacologyAuthor(s): Pallavi Bhattaram, Kyle Jones Inflammation in the synovium is known to mediate joint destruction in several forms of arthritis. Fibroblast-like synoviocytes (FLS) are cells that reside in the synovial lining of joints and are known to be key contributors to inflammation associated with arthritis. FLS are a major source of inflammatory cytokines and catabolic enzymes that promote joint degeneration. We now know that there exists a direct correlation between the signaling pathways that are activated by the pro-inflammatory molecules produced by the FLS, and the severity of joint degeneration in arthritis. Research focused on understanding the signaling pathways that are activated by these pro-inflammatory molecules has led to major advancements in the understanding of the joint pathology in arthritis. Transcription factors (TFs) that act as downstream mediators of the pro-inflammatory signaling cascades in various cell types have been reported to play an important role in inducing the deleterious transformation of the FLS. Interestingly, recent studies have started uncovering that several TFs that were previously reported to play role in embryonic development and cancer, but not known to have pronounced roles in tissue inflammation, can actually play crucial roles in the regulation of the pathological properties of the FLS. In this review, we will discuss reports that have been able to impart novel arthritogenic roles to TFs that are specialized in embryonic development. We also discuss the therapeutic potential of targeting these newly identified regulators of FLS transformation in the treatment of arthritis.Graphical abstractGraphical abstract for this article
  • The Science of Licking Your Wounds: Function of Oxidants in the Innate
           Immune System
    • Abstract: Publication date: Available online 13 March 2019Source: Biochemical PharmacologyAuthor(s): Brian J. Day Animals often lick their wounds to promote healing. Saliva is thought to have healing properties due to it containing many agents that have antimicrobial properties. A number of these components make up the innate immune system’s oxidant generating network. One of these components is the saliva peroxidase also known as lactoperoxidase (LPO). LPO utilizes hydrogen peroxide (H2O2) and the pseudohalide thiocyanate (-SCN) to generate a broad-spectrum antimicrobial oxidant hypothiocyanous acid (HOSCN). HOSCN has antimicrobial activity against viruses, fungi, and bacteria. Although saliva contains the highest levels of -SCN and HOSCN in the body, this network operates in all extracellular fluids in the body including the blood, tears, nasal and lung fluids, gastric fluid, milk and semen. Another unique property of this system is that -SCN can react directly with all hypohalous acids and haloamines to funnel the oxidants to HOSCN that can be selectively detoxified by the host’s thioredoxin reductase but not by pathogen’s thioredoxin reductases due to evolutionary divergence. New understanding of this system many allow us to develop novel approaches to simultaneously treat inflammation while preventing infections.Graphical abstractGraphical abstract for this articleFigure 4. Thiocyanate (-SCN) is the primary determinant of hypothiocyanous acid (HOSCN) formation during inflammation. Under normal physiological levels of chloride, bromide, and -SCN in the plasma there is almost equal amounts of hypochlorous acid (HOCl) and HOSCN formed by myeloperoxidase (MPO). When -SCN transport is limited as observed in cystic fibrosis a larger proportion of HOCl is generated. Pharmacologic administration of -SCN can shift the proportion of HOSCN generated to almost 100% at concentrations around 400μM [22].
  • Andrographolide Derivative Ameliorates Dextran Sulfate Sodium-Induced
           Experimental Colitis in Mice
    • Abstract: Publication date: Available online 13 March 2019Source: Biochemical PharmacologyAuthor(s): Bao-Jian Guo, Zhuyun Liu, Mo-Yu Ding, Feng Li, Mei Jing, Li-Peng Xu, Yu-Qiang Wang, Zai-Jun Zhang, Yitao Wang, Decai Wang, Guo-Chun Zhou, Ying Wang The therapeutic efficacy of immunosuppressive agents has been intensively studied for colitis management. We synthesized a series of andrographolide derivatives and reported their structure-activity-relationship and anti-inflammatory activity in our previous studies. Among these derivatives, compound 3b exhibited the most potent immunosuppressive activity. In the present study, we assessed the efficacy of 3b in dextran sulfate sodium (DSS)-induced model of acute colitis. Compound 3b was administered intragastrically. The therapeutic effect of 3b was evaluated using disease score and immune cell infiltration. The effect of 3b on Toll-like receptor 4/NF-κB and β-catenin signaling was primarily determined by using immunohistochemistry staining and quantitative real-time PCR. The crosstalk between NF-κB and β-catenin signaling was then assessed in HCT-116 cells. Treatment with 3b significantly downregulated the disease activity index and suppressed the histologic evidence of inflammation in DSS-induced model of acute colitis. Compound 3b inhibited proinflammatory cytokine expression at both the serum and transcription levels. Treatment with 3b also upregulated the number of PCNA-positive and goblet cells in the intestinal crypt and the intestinal expression of mRNA levels of β-catenin target genes. β-Catenin level regulation affected the antiinflammation and anti-apoptotic activities of 3b. This study demonstrated that 3b, a novel andrographolide derivative, suppressed inflammation and significantly reversed colitis pathology. The outcome of colitis treatment with an immunosuppressive agent depends upon the intestinal expression and mutation status of β-catenin.Graphical abstractGraphical abstract for this article
  • Seno-suppressive molecules as new therapeutic perspectives in rheumatic
    • Abstract: Publication date: Available online 13 March 2019Source: Biochemical PharmacologyAuthor(s): Yassin Tachikart, Olivier Malaise, Marcus Mumme, Christian Jorgensen, Jean-Marc Brondello Over the past years, through in vitro studies and unique animal models, biologists and clinicians have demonstrated that cellular senescence is at the root of numerous age-related chronic diseases including osteoarthritis and osteoporosis. This non-proliferative cellular syndrome can modify other surrounding tissue-resident cells through the establishment of a deleterious catabolic and inflammatory microenvironment. Targeting these deleterious cells through local or systemic seno-therapeutic agent delivery in pre-clinical models improves dramatically clinical signs and extends health span. In this review, we will summarize the current knowledge on cellular senescence, list the different strategies for identifying seno-suppressive therapeutic agents and their translations to rheumatic diseases.Graphical abstractGraphical abstract for this article
  • Erythromycin acts through the ghrelin receptor to attenuate inflammatory
           responses in chondrocytes and maintain joint integrity
    • Abstract: Publication date: Available online 9 March 2019Source: Biochemical PharmacologyAuthor(s): Tomoya Uchimura, Daisy S. Nakamura, Eric M. Link, Yoshihiko Noguchi, Satoshi Ōmura, Toshiaki Sunazuka, David J. Greenblatt, Li Zeng Osteoarthritis (OA) is a prevalent disease characterized by chronic joint degeneration and low-grade localized inflammation. There is no available treatment to delay OA progression. We report that in human primary articular chondrocytes, erythromycin, a well-known macrolide antibiotic, had the ability to inhibit pro-inflammatory cytokine Interleukin 1β (IL-1β)-induced catabolic gene expression and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. Furthermore, erythromycin inhibited monosodium iodoacetate (MIA)-induced joint inflammation and cartilage matrix destruction in mice, an arthritis model that reflects the inflammatory and cartilage matrix loss aspects of OA. EM900, an erythromycin-derivative lacking antibiotic function, had the same activity as erythromycin in vitro and in vivo, indicating distinct anti-inflammatory and antibiotic properties. Using an antibody against erythromycin, we found erythromycin was present on chondrocytes in a dose-dependent manner. The association of erythromycin with chondrocytes was diminished in ghrelin receptor null chondrocytes, and administration of the ghrelin ligand prevented the association of erythromycin with chondrocytes. Importantly, the anti-inflammatory activity of erythromycin was diminished in ghrelin receptor null chondrocytes. Moreover, erythromycin could not exert its chondroprotective effect in ghrelin receptor null mice, and the loss of ghrelin receptor further augmented joint damage upon MIA-injection. Therefore, our study identified a novel pharmacological mechanism for how erythromycin exerts its chondroprotective effect. This mechanism entails ghrelin receptor signaling, which is necessary for alleviating inflammation and joint destruction.Graphical abstractGraphical abstract for this article
  • Role of mitochondrial dysfunction on rheumatic diseases
    • Abstract: Publication date: Available online 9 March 2019Source: Biochemical PharmacologyAuthor(s): Carlos Vaamonde-García, María J López-Armada Rheumatic and musculoskeletal diseases are a heterogeneous group of disorders affecting joint tissues and in some cases even organs, some of them being among the most common diseases worldwide. Mitochondria are the organelles considered as powerhouse of cells providing energy to the organism mainly through oxidative phosphorylation. However, mitochondria are also involved in crucial pathways responsible for maintaining cell physiology, such as the activation of metabolic and survival signaling, and innate and adaptive immune response. As consequence of the pivotal role of mitochondria in cell homeostasis, an impairment of mitochondrial function has been associated with activation of pathological events, including oxidative stress and subsequently damaged protein and DNA, deregulation of programmed cell death, and over-activation of inflammatory responses modulated by redox-sensitive signaling or direct activation of the inflammasome. Thus, a growing amount of evidence emphasizes the role of mitochondria in aging and inflammatory-related diseases, including rheumatic disorders. In this regard, emerging findings suggest that targeting of the pathways involved in the maintenance of mitochondrial metabolism may control cell homeostasis, and in turn delay ageing and prevent or improve articular pathologies. In this review we will focus on the importance of mitochondria in metabolic homeostasis of articular cells, as well as their influence on the activation of pathological signaling pathways, as a result of a genetic predisposition, damage, decline or impairment of their function. Finally, we will discuss some of the most important evidences of involvement of mitochondria in the onset and progression of rheumatic diseases.Graphical abstractGraphical abstract for this article
  • Regulation of neutrophil pro-inflammatory functions sheds new light on the
           pathogenesis of rheumatoid arthritis
    • Abstract: Publication date: Available online 9 March 2019Source: Biochemical PharmacologyAuthor(s): N. Jung, J.-L. Bueb, F. Tolle, S. Bréchard For more than two centuries now, rheumatoid arthritis (RA) is under investigation intending to discover successful treatment. Despite decades of scientific advances, RA is still representing a challenge for contemporary medicine. Current drug therapies allow to improve significantly the quality of life of RA patients; however, they are still insufficient to reverse tissue injury and are often generating side-effects. The difficulty arises from the considerable fluctuation of the clinical course of RA among patients, making the predictive prognosis difficult. More and more studies underline the profound influence of the neutrophil multifaceted functions in the pathogenesis of RA. This renewed interest in the complexity of neutrophil functions in RA offers new exciting opportunities for valuable therapeutic targets as well as for safe and well-tolerated RA treatments. In this review, we aim to update the recent findings on the multiple facets of neutrophils in RA, in particular their impact in promoting the RA-based inflammation through the release of the cytokine-like S100A8/A9 protein complex, as well as the importance of NETosis in the disease progression and development. Furthermore, we delve into the complex question of neutrophil heterogeneity and plasticity and discuss the emerging role of miRNAs and epigenetic markers influencing the inflammatory response of neutrophils in RA and how they could constitute the starting point for novel attractive targets in RA therapy.Graphical abstractGraphical abstract for this article
  • Oncogenic EP300 can be targeted with inhibitors of aldo-keto reductases
    • Abstract: Publication date: Available online 9 March 2019Source: Biochemical PharmacologyAuthor(s): Zimam Mahmud, Muhammad Asaduzzaman, Uttom Kumar, Nahal Masrour, Roman Jugov, R. Charles Coombes, Sami Shousha, Yunhui Hu, Eric W.-F. Lam, Ernesto Yagüe E-cadherin transcriptional activator EP300 is down-regulated in metaplastic breast carcinoma, a rare form of triple negative and E-cadherin-negative aggressive breast cancer with a poor clinical outcome. In order to shed light on the regulation of E-cadherin by EP300 in breast cancer we analyzed by immunohistochemistry 41 cases of invasive breast cancer with both E-cadherinhigh and E-cadherinlow expression levels, together with 20 non-malignant breast tissues. EP300 and E-cadherin showed a positive correlation in both non-malignant and cancer cases and both markers together were better predictors of lymph node metastasis than E-cadherin alone. These data support a metastasis suppressor role for EP300 in breast cancer. However, some reports suggest an oncogenic role for EP300. We generated a breast cancer cell model to study E-cadherin-independent effects of EP300 by over-expressing of EP300 in HS578T cells which have E-cadherin promoter hypermethylated. In this cell system, EP300 led to up-regulation of mesenchymal (vimentin, Snail, Slug, Zeb1) and stemness (ALDH+ and CD44high/CD24low) markers, increases in migration, invasion, anchorage-independent growth and drug resistance. Genome-wide expression profiling identified aldo-keto reductases AKR1C1-3 as effectors of stemness and drug resistance, since their pharmacological inhibition with flufenamic acid restored both doxorubicin and paclitaxel sensitivity and diminished mammosphere formation. Thus, in cells with a permissive E-cadherin promoter, EP300 acts as a tumour / metastasis supressor by up-regulating E-cadherin expression, maintenance of the epithelial phenotype and avoidance of an epithelial-to-mesenchymal transition. In cells in which the E-cadherin promoter is hypermethylated, EP300 functions as an oncogene via up-regulation of aldo-keto reductases. This offers the rationale of using current aldo-keto reductase inhibitors in breast cancer treatment.Graphical abstractGraphical abstract for this article
  • An overview of the multifaceted roles of miRNAs in gastric cancer:
           Spotlight on novel biomarkers and therapeutic targets
    • Abstract: Publication date: Available online 9 March 2019Source: Biochemical PharmacologyAuthor(s): Xu Wu, Jing Shen, Zhangang Xiao, Jing Li, Yueshui Zhao, Qijie Zhao, Chi Hin Cho, Mingxing Li MicroRNAs (miRNAs) are a group of small non-coding RNAs that have displayed strong association with gastric cancer (GC). Through the repression of target mRNAs, miRNAs regulate many biological pathways that are involved in cell proliferation, apoptosis, migration, invasion, metastasis as well as drug resistance. The detection of miRNAs in tissues and in body fluids emerges as a promising method in the diagnosis and prognosis of GC, due to their unique expression pattern in correlation with GC. Notably, miRNAs are also identified as potential therapeutic targets for GC therapy. The present review is thus to highlight the multifaceted roles of miRNAs in GC and in GC therapies, which would give indications for future researches.Graphical abstractGraphical abstract for this article
  • Penicisulfuranol A, A Novel C-terminal Inhibitor Disrupting Molecular
           Chaperone Function of Hsp90 Independent of ATP Binding Domain
    • Abstract: Publication date: Available online 8 March 2019Source: Biochemical PharmacologyAuthor(s): Jiajia Dai, Ao Chen, Meilin Zhu, Xin Qi, Wei Tang, Ming Liu, Dehai Li, Qianqun Gu, Jing Li The goal of this study is to explore the mechanism of a heat shock protein 90 (Hsp90) C-terminal inhibitor, Penicisulfuranol A (PEN-A), for cancer therapy. Penicisulfuranol A (PEN-A) was produced by a mangrove endophytic fungus Penicillium janthinellum and had a new structure with a rare 3H-spiro [benzofuran-2, 2'-piperazine] ring system. PEN-A caused depletion of multiple Hsp90 client proteins without induction of heat shock protein 70 (Hsp70). Subsequently, it induced apoptosis and inhibited xerograph tumor growth of HCT116 cells in vitro and in vivo. Mechanism studies showed that PEN-A was bound to C-terminus of Hsp90 at the binding site different from ATP binding domain. Therefore, it inhibited dimerization of Hsp90 C-terminus, depolymerization of ADH protein by C-terminus of Hsp90, and interaction of co-chaperones with Hsp90. These inhibitory effects of PEN-A were similar to those of novobiocin, an inhibitor binding to interaction site for ATP of C-terminus of Hsp90. Furthermore, our study revealed that disulfide bond was essential moiety for inhibition activity of PEN-A on Hsp90. This suggested that PEN-A may be bound to cysteine residues near amino acid region which was responsible for dimerization of Hsp90. All results indicate that PEN-A is a novel C-terminal inhibitor of Hsp90 and worthy for further study in the future not only for drug development but also for unraveling the bioactivities of Hsp90.Graphical abstractGraphical abstract for this article
  • Chondrocyte dedifferentiation and osteoarthritis (OA)
    • Abstract: Publication date: Available online 7 March 2019Source: Biochemical PharmacologyAuthor(s): Charlier Edith, Deroyer Céline, Ciregia Federica, Malaise Olivier, Neuville Sophie, Plener Zelda, Malaise Michel, de Seny Dominique Osteoarthritis (OA) is a degenerative joint disease characterized by progressive cartilage degradation but also synovial membrane inflammation, osteophyte formation and subchondral bone sclerosis. Medical care is mainly based on alleviating pain symptoms, but to date, no effective drug can stop the disease progression. Cartilage is a tissue composed of only one cell type, chondrocytes, wrapped in a collagen rich extracellular matrix they synthesize. Chondrocytes can adopt different phenotypes in vivo and in vitro, defined by the collagen type they produce. Isolated from their matrix, chondrocytes present the particularity to dedifferentiate, producing fibroblastic type I and III collagens. With OA onset, chondrocytes undergo multiple changes, in terms of proliferation, viability, but also secretory profile. The acquisition of a hypertrophic phenotype (producing aberrant type X collagen and catabolic MMP-13 protease) by chondrocytes is well documented and contributes to OA development. However, it is increasingly believed that chondrocytes rather acquire a variety of degenerated phenotypes at the onset of OA, including a “dedifferentiated-like” phenotype that might also contribute to OA progression. In this review, we will (i) present molecular knowledge underlying dedifferentiation process, (ii) emphasize connections between dedifferentiation and OA and (iii) consider OA therapeutic strategies aiming at the maintenance of chondrogenic phenotype.Graphical abstractGraphical abstract for this article
  • The PDE4 inhibitor CHF6001 modulates pro-inflammatory cytokines,
           chemokines and Th1- and Th17-polarizing cytokines in human dendritic cells
    • Abstract: Publication date: Available online 6 March 2019Source: Biochemical PharmacologyAuthor(s): Veronica Gianello, Valentina Salvi, Carmen Parola, Nadia Moretto, Fabrizio Facchinetti, Maurizio Civelli, Gino Villetti, Daniela Bosisio, Silvano Sozzani Phosphodiesterase 4 (PDE4) inhibitors are used to treat autoimmune and inflammatory diseases, such as psoriasis and chronic obstructive pulmonary disease (COPD). CHF6001 is a novel, potent and selective inhaled PDE4 inhibitor in development for the treatment of COPD. When tested in vitro on human dendritic cells (DCs), CHF6001 decreased the release of pro-inflammatory cytokines (TNFα and IL-6), chemokines (CXCL8, CCL3, CXCL10 and CCL19) and of Th1- and Th17-polarizing cytokines (IL-12, IL-23 and IL-1β). In contrast to β -methasone, a reference steroid anti-inflammatory drug, CHF6001 increased the secretion of CCL22, a Th2 recruiting chemokine, and the expression of the lymph node homing receptor CCR7. Accordingly, the migration of DCs to CCR7 ligands was increased, while migration to pro-inflammatory chemokines was decreased. Of note, the action of CHF6001 was apparently mediated by a promoter-specific decrease in NF-κB p65 recruitment, independent of perturbation of LPS signalling or NF-κB nuclear translocation. Our results indicate that CHF6001 can modulate DC pro-inflammatory Th1/Th17 polarizing potential by fine tuning the transcriptional activity of the master inflammatory transcription factor NF-κB. Therefore, CHF6001 may prove useful to control Th1/Th17-polarized inflammatory diseases such as COPD.Graphical abstractGraphical abstract for this article
  • Proteasomal inhibition attenuates craniofacial malformations in a
           zebrafish model of Treacher Collins Syndrome
    • Abstract: Publication date: Available online 5 March 2019Source: Biochemical PharmacologyAuthor(s): Mauco Gil Rosas, Agustín Lorenzatti, Mauro S. Porcel de Peralta, Nora B. Calcatera, Gabriela Coux Treacher Collins Syndrome (TCS) is a congenital disease characterized by defects in the craniofacial skeleton and absence of mental alterations. Recently we modelled TCS in zebrafish (Danio rerio) embryos through the microinjection of Morpholino® oligonucleotides blocking the translation of the ortholog of the main causative gene (TCOF1). We showed that Cnbp, a key cytoprotective protein involved in normal rostral head development, was detected in lower levels (without changes in its mRNA expression) in TCS-like embryos. As previous reports suggested that Cnbp is degraded through the proteasomal pathway, we tested whether proteasome inhibitors (MG132 and Bortezomib (Velcade®, Millennium laboratories)) were able to ameliorate cranial skeleton malformations in TCS. Here we show that treatment with both proteasome inhibitors produced a robust craniofacial cartilage phenotype recovery. This recovery seems to be consequence of a decreased degradation of Cnbp in TCS-like embryos. Critical TCS manifestations, such as neuroepithelial cell death and cell redox imbalance were attenuated. Thus, proteasome inhibitors may offer an opportunity for TCS molecular and phenotypic manifestation’s prevention. Although further development of new safe inhibitors compatible with administration during pregnancy is required, our results encourage this therapeutic approach.Graphical abstractGraphical abstract for this article
  • The modulation of acid-sensing ion channel 1 by PcTx1 is pH-, subtype- and
           species-dependent: importance of interactions at the channel subunit
           interface and potential for engineering selective analogues
    • Abstract: Publication date: Available online 5 March 2019Source: Biochemical PharmacologyAuthor(s): Ben Cristofori-Armstrong, Natalie J. Saez, Irène R. Chassagnon, Glenn F. King, Lachlan D. Rash Acid-sensing ion channels (ASICs) are primary acid sensors in the mammalian nervous system that are activated by protons under conditions of local acidosis. They have been implicated in a range of pathologies including ischemic stroke (ASIC1a subtype) and peripheral pain (ASIC1b and ASIC3). Although the spider venom peptide PcTx1 is the best-studied ASIC modulator and is neuroprotective in rodent models of ischemic stroke, little experimental work has been done to examine its molecular interaction with human ASIC1a or the off-target ASIC1b. The complementary face of the acidic pocket binding site of PcTx1 is where these channels differ in sequence. We show here that although PcTx1 is 10-fold less potent at human ASIC1a than the rat channel, the apparent affinity for the two channels is comparable. We examined the pharmacophore of PcTx1 for human ASIC1a and rat ASIC1b, and show that inhibitory and stimulatory effects at each ASIC1 variant is driven mostly by a shared set of core peptide pharmacophore residues that bind to the thumb domain, while peptide residues that interact with the complementary face of the biding site underlie species and subtype-dependent differences in activity that may allow manipulation of ASIC1 variant selectivity. Finally, the stimulatory effect of PcTx1 on rat ASIC1a when applied under mildly alkaline pH correlates with low receptor occupancy. These new insights into the interactions between PcTx1 with ASIC1 subtypes demonstrates the complexity of its mechanism of action, and highlights important implications to consider when using PcTx1 as a pharmacological tool to study ASIC function.Graphical abstractGraphical abstract for this article
  • Novel benzoxanthene lignans that favorably modulate lipid mediator
           biosynthesis: a promising pharmacological strategy for anti-inflammatory
    • Abstract: Publication date: Available online 2 March 2019Source: Biochemical PharmacologyAuthor(s): Jana Gerstmeier, Christian Kretzer, Simone Di Micco, Laura Miek, Hannah Butschek, Vincenza Cantone, Rossella Bilancia, Roberta Rizza, Fabiana Troisi, Nunzio Cardullo, Corrado Tringali, Armando Ialenti, Antonietta Rossi, Giuseppe Bifulco, Oliver Werz, Simona Pace Lipid mediators (LM) encompass pro-inflammatory prostaglandins (PG) and leukotrienes (LT) but also specialized pro-resolving mediators (SPM) which display pivotal bioactivities in health and disease. Pharmacological intervention with inflammatory disorders such as osteoarthritis and rheumatoid arthritis commonly employs anti-inflammatory drugs that can suppress PG and LT formation, which however, possess limited effectiveness and side effects. Here, we report on the discovery and characterization of the two novel benzoxanthene lignans 1 and 2 that modulate select LM biosynthetic enzymes enabling the switch from pro-inflammatory LT to SPM biosynthesis as potential pharmacological strategy to intervene with inflammation. In cell-free assays, compound 1 and 2 inhibit microsomal prostaglandin E2 synthase-1 and leukotriene C4 synthase (IC50 ∼ 0.6-3.4 µM) and potently interfere with 5-lipoxygenase (5-LOX), the key enzyme in LT biosynthesis (IC50 = 0.04 and 0.09 µM). In human neutrophils, monocytes and M1 and M2 macrophages, compound 1 and 2 efficiently suppress LT biosynthesis (IC50 < 1 µM), accompanied by elevation of 15-LOX-derived LM including SPM. In zymosan-induced murine peritonitis, compound 1 and 2 ameliorated self-limited inflammation along with suppression of early LT formation and elevation of subsequent SPM biosynthesis in vivo. Together, these novel benzoxanthene lignans promote the LM class switch from pro-inflammatory towards pro-resolving LM to terminate inflammation, suggesting their suitability as novel leads for pharmacotherapy of arthritis and related inflammatory disorders.Graphical abstractGraphical abstract for this article
  • The FICI paradigm: correcting flaws in antimicrobial in vitro synergy
           screens at their inception
    • Abstract: Publication date: Available online 2 March 2019Source: Biochemical PharmacologyAuthor(s): Marta Gómara, Santiago Ramón-García Antibiotics have become the corner stone of modern medicine. However, our society is currently facing one of the greatest challenges of its time: the emergence of antimicrobial resistance. It is estimated that if no new therapies are implemented by 2050, 10 million people will die worldwide every year as a result of infections caused by bacteria resistant to current antibiotics; new antimicrobials are thus urgently needed. However, drug development is a tedious and very costly endeavor of hundreds of millions that can take up to 15-20 years from the bench discovery to the bedside. Under this scenario, drug repurposing, which consists in identifying new uses for old, clinically approved drugs, has gathered momentum within the pharmaceutical industry. Because most of these drugs have safety and toxicity information packages available, clinical evaluation could be done in a much shorter period than standard timelines. Synergistic combinations of these clinically approved drugs could also be a promising approach to identify novel antimicrobial therapies that might provide rational choices of available drugs to shorten treatment, increase efficacy, reduce toxicity, prevent resistance and treat infections caused by drug-resistant strains. However, although simple in its conception, translating results from in vitro synergy screens into in vivo efficacy or the clinical practice has proven to be a paramount challenge.In this Commentary, we will discuss common flaws at the inception of synergy research programs, with a special focus on the use of the Fractional Inhibitory Concentration Index (FICI), and evaluate potential interventions that can be made at different developmental pre-clinical stages in order to improve the odds of translation from in vitro studies.Graphical abstractGraphical abstract for this article
  • Functional and molecular characterization of UDP-glucuronosyltransferase 2
           family in cynomolgus macaques
    • Abstract: Publication date: Available online 2 March 2019Source: Biochemical PharmacologyAuthor(s): Yasuhiro Uno, Rika Takahira, Norie Murayama, Shunsuke Onozeki, Shu Kawamura, Shotaro Uehara, Yoshinori Ikenaka, Mayumi Ishizuka, Shinichi Ikushiro, Hiroshi Yamazaki UDP-glucuronosyltransferases (UGTs) are essential enzymes metabolizing endogenous and exogenous chemicals. However, characteristics of UGTs have not been fully investigated in molecular levels of cynomolgus macaques, one of non-human primates widely used in preclinical drug metabolism studies. In this study, three UGT2A cDNAs (UGT2A1, 2A2, and 2A3) were isolated and characterized along with seven UGT2Bs previously identified in cynomolgus macaques. Several transcript variants were found in cynomolgus UGT2A1 and UGT2A2, like human orthologs. Cynomolgus UGT2A and UGT2B amino acid sequences were highly identical (87-96%) to their human counterparts. By phylogenetic analysis, all these cynomolgus UGT2s were more closely clustered with their human homologs than with dog, rat, or mouse UGT2s. Especially, UGT2As showed orthologous relationships between humans and cynomolgus macaques. All the cynomolgus UGT2 mRNAs were expressed in livers, jejunum, and/or kidneys abundantly, except that UGT2A1 and UGT2A2 mRNAs were predominantly expressed in nasal mucosa, like human UGT2s. UGT2A and UGT2B genes together form a gene cluster in the cynomolgus and human genome. Among the seven cynomolgus UGT2Bs heterologously expressed in yeast, UGT2B9 and UGT2B30 showed activities in estradiol 17-O-glucuronidation and morphine 3-O-glucuronidation but did not show activities in estradiol 3-O-glucuronidation, similar to human UGT2Bs. In liver microsomes, cynomolgus macaques showed higher estradiol 17-O-glucuronidase and morphine 3-O-glucuronidase activities than humans, suggesting functional activities of the responsible UGT2B enzymes in cynomolgus macaques. Therefore, cynomolgus UGT2s had overall molecular similarities to human UGT2s, but also showed some differences in UGT2B enzyme properties.Graphical abstractGraphical abstract for this article
  • Pharmacological effectors of GRP78 chaperone in cancers
    • Abstract: Publication date: Available online 1 March 2019Source: Biochemical PharmacologyAuthor(s): Christian Bailly, Michael J. Waring The protein chaperone GRP78 is a master regulator of endoplasmic reticulum (ER) functions and is frequently over-expressed at the surface of cancer cells where it contributes to chemo-resistance. It represents a well-studied ER stress marker but an under-explored target for new drug development. This review aims to untangle the structural and functional diversity of GRP78 modulators, covering over 130 natural products, synthetic molecules, specific peptides and monoclonal antibodies that target GRP78. Several approaches to promote or to incapacitate GRP78 are presented, including the use of oligonucleotides and specific cell-delivery peptides often conjugated to cytotoxic payloads to design GRP78-targeted therapeutics. A repertoire of drugs that turn on/off GRP78 is exposed, including molecules which bind directly to GRP78, principally to its ATP site. There exist many options to regulate positively or negatively the expression of the chaperone, or to interfere with its cellular trafficking. This review provides a molecular cartography of GRP78 pharmacological effectors and adds weight to the notion that GRP78 repressors could represent promising anticancer therapeutics, notably as regards limiting chemo-resistance of cancer cells. The potential of GRP78-targeting drugs in other therapeutic modalities is also evoked.Graphical abstractGraphical abstract for this article
  • Osteoarthritis phenotypes and novel therapeutic targets
    • Abstract: Publication date: Available online 1 March 2019Source: Biochemical PharmacologyAuthor(s): Willem Evert Van Spil, Olga Kubassova, Mikael Boesen, Anne-Christine Bay-Jensen, Ali Mobasheri The success of disease-modifying osteoarthritis drug (DMOAD) development is still elusive. While there have been successes in preclinical and early clinical studies, phase 3 clinical trials have failed so far and there is still no approved, widely available DMOAD on the market. The latest research suggests that, among other causes, poor trial outcomes might be explained by the fact that osteoarthritis (OA) is a heterogeneous disease with distinct phenotypes. OA trials might be more successful if they would address and target a specific phenotype.The increasing availability of advanced techniques to detect particular OA characteristics expands the possibilities to distinguish between such potential OA phenotypes. Magnetic resonance imaging is among the key imaging techniques to stratify and monitor patients with changes in bone, cartilage and inflammation. Biochemical markers have mainly used as secondary parameters and could further delineate phenotypes. Moreover, post-hoc analyses of trial data have suggested the existence of distinct pain phenotypes and their relevance in the design of clinical trials.Although ongoing work in the field supports the concept of OA heterogeneity, this has not yet resulted in more effective treatment options. This paper reviews the current knowledge about potential OA phenotypes and suggests that combining patient clinical data, quantitative imaging, biochemical markers and utilizing data-driven approaches in patient selection and efficacy assessment will allow for more successful development of effective DMOADs.Graphical abstractGraphical abstract for this article
  • Cyclin-dependent kinase inhibitors AZD5438 and R547 show potential for
           enhancing efficacy of daunorubicin-based anticancer therapy: interaction
           with carbonyl-reducing enzymes and ABC transporters
    • Abstract: Publication date: Available online 28 February 2019Source: Biochemical PharmacologyAuthor(s): Ales Sorf, Eva Novotna, Jakub Hofman, Anselm Morell, Frantisek Staud, Vladimir Wsol, Martina Ceckova Daunorubicin (DAUN) has served as an anticancer drug in chemotherapy regimens for decades and is still irreplaceable in treatment of acute leukemias. The therapeutic outcome of DAUN-based therapy is compromised by its cardiotoxicity and emergence of drug resistance. This phenomenon is often caused by pharmacokinetic mechanisms such as efflux of DAUN from cancer cells through ATP-binding cassette (ABC) transporters and its conversion to less cytostatic but more cardiotoxic daunorubicinol (DAUN-OL) by carbonyl reducing enzymes (CRE). Here we aimed to investigate, whether two cyclin-dependent kinase inhibitors, AZD5438 and R547, can interact with these pharmacokinetic mechanisms and reverse DAUN resistance. Using accumulation assays, we revealed AZD5438 as potent inhibitor of ABCC1 showing also weaker inhibitory effect to ABCB1 and ABCG2. Combination index analysis, however, shown that inhibition of ABCC1 does not significantly contribute to synergism between AZD5438 and DAUN in MDCKII-ABCC1 cells, suggesting predominant role of other mechanism. Using pure recombinant enzymes, we found both tested drugs to inhibit CREs with aldo-keto reductase 1C3 (AKR1C3). This interaction was further confirmed in transfected HCT-116 cells. Moreover, these cells were sensitized to DAUN by both compounds as Chou-Talalay combination index analysis showed synergism in AKR1C3 transfected HCT-116, but not in empty vector transfected control cell line. In conclusion, we propose AZD5438 and R547 as modulators of DAUN resistance that can prevent AKR1C3-mediated DAUN biotransformation to DAUN-OL. This interaction could be beneficially exploited to prevent failure of DAUN-based therapy as well as the undesirable cardiotoxic effect of DAUN-OL.Graphical abstractGraphical abstract for this article
  • Na+/Ca2+ Exchangers: unexploited opportunities for
           cancer therapy'
    • Abstract: Publication date: Available online 28 February 2019Source: Biochemical PharmacologyAuthor(s): Tiago Rodrigues, Gabriela Nohemi Nunez Estevez, Ivarne Luis dos Santos Tersariol Calcium is a well-studied ion that acts as a cofactor in several reactions and as intracellular second messenger. It plays crucial roles in living cells by regulating several processes from cell division to death. The disruption of Ca2+ homeostasis is related to cell and tissue damage and it is involved in several pathological conditions and diseases, including cancer. Tumor cells exhibit several molecular features in relation to normal cells in order to acquire proliferative and survival advantages, and Ca2+ signaling is directly or indirectly involved in these pathways. Thus, changes in the expression of Ca2+ channels and pumps are frequently described in some cancers, including transient receptor potential (TRP) family channels, store- and voltage-gated Ca2+ channels, store release channels, and Ca2+ ATPases. Although the sodium/calcium exchanger (Na+/Ca2+ exchanger; NCX) and the therapeutic potential of its inhibitors have been extensively studied in heart diseases, there are few studies about the molecular and functional aspects of NCX in cancer. Here, the current knowledge about NCX in cancer will be reviewed and possible strategies to target NCX for cancer therapy will be discussed.Graphical abstractGraphical abstract for this article
  • Inhibition of MMPs and ADAM/ADAMTS
    • Abstract: Publication date: Available online 28 February 2019Source: Biochemical PharmacologyAuthor(s): Charles J. Malemud Matrix metalloproteinases (MMPs), A Disintegrin and Metalloproteinase (ADAM) and A Disintegrin and Metalloproteinase with Thrombospondin Motif (ADAMTS) are zinc-dependent endopeptidases that play a critical role in the destruction of extracellular matrix proteins and, the shedding of membrane-bound receptor molecules in various forms of arthritis and other diseases. Under normal conditions, MMP, ADAM and ADAMTS gene expression aids in the maintenance of homeostasis. However, in inflamed synovial joints characteristic of rheumatoid arthritis and osteoarthritis. MMP, ADAM and ADAMTS production is greatly increased under the influence of pro-inflammatory cytokines. Analyses based on medicinal chemistry strategies designed to directly inhibit the activity of MMPs have been largely unsuccessful when these MMP inhibitors were employed in animal models of rheumatoid arthritis and osteoarthritis. This is despite the fact that these MMP inhibitors were largely able to suppress pro-inflammatory cytokine-induced MMP production in vitro. A focus on ADAM and ADAMTS inhibitors has also been pursued. Thus, recent progress has identified the “sheddase” activity of ADAMs as a viable target and the development of GW280264X is an experimental ADAM17 inhibitor. Of note, a monoclonal antibody, GLPG1972, developed as an ADAMTS-5 inhibitor, entered a Phase I OA clinical trial. However, the failure of many of these previously developed inhibitors to move beyond the preclinical testing phase has required that novel strategies be developed that are designed to suppress both MMP, ADAM and ADAMTS production and activity.Graphical abstractGraphical abstract for this article
  • Emerging therapeutic agents in osteoarthritis
    • Abstract: Publication date: Available online 28 February 2019Source: Biochemical PharmacologyAuthor(s): María José Alcaraz, María Isabel Guillén, María Luisa Ferrándiz Osteoarthritis (OA) is the most common joint disorder and a leading cause of disability. Current treatments for OA can improve symptoms but do not delay the progression of disease. In the last years, much effort has been devoted to developing new treatments for OA focused on pain control, inflammatory mediators or degradation of articular tissues. Although promising results have been obtained in ex vivo studies and animal models of OA, few of these agents have completed clinical trials. Available clinical data support the interest of nerve growth factor as a target in pain control as well as the disease-modifying potential of inhibitors of Wnt signaling or catabolic enzymes such as aggrecanases and cathepsin K, and anabolic strategies like fibroblast growth factor-18 or cellular therapies. Carefully controlled studies in patients selected according to OA phenotypes and with a long follow-up will help to confirm the relevance of these new approaches as emerging therapeutic treatments in OA.Graphical abstractGraphical abstract for this article
  • Micro-RNAs in inflammatory arthritis: from physiopathology to diagnosis,
           prognosis and therapeutic opportunities
    • Abstract: Publication date: Available online 27 February 2019Source: Biochemical PharmacologyAuthor(s): Aurélie Najm, Frédéric Blanchard, Benoit Le Goff Micro-RNAs are an area of research exponentially expanding over the past years. These small sequences of 20 to 22 nucleotides have a strong role as post-transcriptional regulators of gene expression. Inflammatory arthritis pathophysiology involves various key players from innate to adaptive immunity, as well as various signalling pathways of inflammation. In this review, we discuss how micro-RNAs are involved in rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis and juvenile inflammatory arthritis, from pre-clinical phases to established diseases. We describe mi-RNAs key roles in fibroblast like synoviocytes migration, proliferation, apoptosis and cytokine production, in macrophages polarization, as well as in B cells and T cell proliferation and differentiation, with a special emphasis on Treg/Th17 imbalance. We finally discuss the application of these findings in pre-clinical models and highlight opportunities and limits of a therapeutic approach using mi-RNAs agonists or antagonists.Graphical abstractMi-RNAs target players of key signalling pathways and allow a better understanding of arthritis pathophysiology. Moreover, mi-RNAs constitute a wide source of therapeutic agents to be potentially used in arthritis.Graphical abstract for this article
  • Targeting Chronic Innate Inflammatory Pathways, the Main Road to
           Prevention of Osteoarthritis Progression
    • Abstract: Publication date: Available online 27 February 2019Source: Biochemical PharmacologyAuthor(s): Gabriel Herrero-Beaumont, Sandra Pérez-Baos, Olga Sánchez-Pernaute, Jorge A Roman-Blas, Ana Lamuedra, Raquel Largo Osteoarthritis (OA) is a chronic joint disease characterized by cartilage degradation, osteophyte formation, subchondral bone sclerosis, and synovitis. Systemic factors such as obesity and the components of the metabolic syndrome seem to contribute to its progression. Breakdown of cartilage ensues from an altered balance between mechanical overload and its absorption by this tissue. There is in this context a status of persistent local inflammation by means of the chronic activation of innate immunity. A broad variety of danger-associated molecular patterns inside OA joint are able to activate pattern recognition receptors, mainly TLR (toll-like receptor) 2 and 4, which are overexpressed in the OA cartilage. Chronic activation of innate immune responses in chondrocytes results in a robust production of pro-inflammatory cytokines and chemokines, as well as of tissue-destructive enzymes, downstream of NF-κB and MAPK (mitogen activated protein kinase) dependent pathways. Besides, the toxic effects of an excess of glucose and/or fatty acids, which share the same pro-inflammatory intracellular signalling pathways, may add fuel to the fire. Not only high concentrations of glucose can render cells prone to inflammation, but also AGEs (advanced glycation end products) are integrated into the TLR signalling network through their own innate immune receptors. Considering these mechanisms, we argue for the control of both primary inflammation and proteolytic catabolism as a preventive strategy in OA, instead of focusing treatment on the enhancement of anabolic responses. Even though this approach would not return to normal already degraded cartilage, it nonetheless might avoid damage extension to the surrounding tissue.Graphical abstractGraphical abstract for this article
  • Cannabinoid derivatives acting as dual PPARγ/CB2 agonists as therapeutic
           agents for Systemic Sclerosis
    • Abstract: Publication date: Available online 27 February 2019Source: Biochemical PharmacologyAuthor(s): Adela García-Martín, Martín Garrido-Rodríguez, Carmen Navarrete, Diego Caprioglio, Belén Palomares, Jim DeMesa, Alain Rollland, Giovanni Appendino, Eduardo Muñoz The endocannabinoid system (ECS) may play a role in the pathophysiology of systemic sclerosis (SSc). Cannabinoids acting as dual PPARγ/CB2 agonists, such as VCE-004.8 and Ajulemic acid (AjA), have been shown to alleviate skin fibrosis and inflammation in SSc models. Since both compounds are being tested in humans, we compared their activities in the bleomycin (BLM) SSc model. Specifically, the pharmacotranscriptomic signature of the compounds was determined by RNA-Seq changes in the skin of BLM mice treated orally with AjA or EHP-101, a lipidic formulation of VCE-004.8. While both compounds down-regulated the expression of genes involved in the inflammatory and fibrotic components of the disease and the pharmacotranscriptomic signatures were similar for both compounds in some pathways, we found key differences between the compounds in vasculogenesis. Additionally, we found 28 specific genes with translation potential by comparing with a list of human scleroderma genes. Immunohistochemical analysis revealed that both compounds prevented fibrosis, collagen accumulation and Tenascin C (TNC) expression. The endothelial CD31+/CD34+ cells and telocytes were reduced in BLM mice and restored only by EHP-101 treatment. Finally, differences were found in plasmatic biomarker analysis; EHP-101, but not AjA, enhanced the expression of some factors related to angiogenesis and vasculogenesis. Altogether the results indicate that dual PPARγ/CB2 agonists qualify as a novel therapeutic approach for the treatment of SSc and other fibrotic diseases. EHP-101 demonstrated unique mechanisms of action related to the pathophysiology of SSc that could be beneficial in the treatment of this complex disease without current therapeutic options.Graphical abstractGraphical abstract for this article
  • Suppressive activities of KC1-3 on HMGB1-mediated septic responses
    • Abstract: Publication date: Available online 26 February 2019Source: Biochemical PharmacologyAuthor(s): Wonhwa Lee, O. Yuseok, Changhun Lee, So Yeon Jeong, Jee-Hyun Lee, Moon-Chang Baek, Gyu-Yong Song, Jong-Sup Bae In the present study, several decursin analogues (KC1–3) were synthesized and evaluated in terms of their anti-septic activities on high mobility group box 1 (HMGB1)-mediated septic responses and survival rate in a mouse model of sepsis. KC1 and KC3, but not KC2, significantly reduced HMGB1 release in lipopolysaccharide (LPS)-activated human umbilical vein endothelial cells (HUVECs) and attenuated the cecal ligation and puncture (CLP)-induced release of HMGB1. Additionally, in vitro analyses revealed that KC1 and KC3 both alleviated HMGB1-mediated vascular disruptions and inhibited hyperpermeability in mice, and in vivo analyses revealed that KC1 and KC3 reduced sepsis-related mortality and tissue injury. Taken together, the present results suggest that KC1 and KC3 both reduced HMGB1 release and septic mortality and, thus, may be useful for the treatment of sepsis.Graphical abstractGraphical abstract for this article
  • Andrographolide and its potent derivative exhibit anticancer effects
           against imatinib-resistant chronic myeloid leukemia cells by
           downregulating the Bcr-Abl oncoprotein
    • Abstract: Publication date: Available online 26 February 2019Source: Biochemical PharmacologyAuthor(s): Hsin-Chia Liao, Yi-Ju Chou, Ching-Cheng Lin, Sheng-Hung Liu, Audrey Oswita, Yi-Long Huang, Ying-Lien Wang, Jia-Ling Syu, Chung-Ming Sun, Chuen-Miin Leu, Chao-Hsiung Lin, Shu-Ling Fu Chronic myelogenous leukemia (CML) is clinically treated with imatinib, which inhibits the kinase activity of the Bcr-Abl oncoprotein. However, imatinib resistance remains a common clinical issue. Andrographolide, the major compound of the medicinal plant Andrographis paniculata, was reported to exhibit anticancer activity. In this study, we explored the therapeutic potential of andrographolide and its derivative, NCTU-322, against both imatinib-sensitive and imatinib-resistant human CML cell lines. Both andrographolide and NCTU-322 downregulated the Bcr-Abl oncoprotein in imatinib-resistant CML cells through an Hsp90-dependent mechanism similar to that observed in imatinib-sensitive CML cells. In addition, NCTU-322 had stronger effects than andrographolide on downregulation of Bcr-Abl oncoprotein, induction of Hsp90 cleavage and cytotoxicity of CML cells. Notably, andrographolide and NCTU-322 could induce differentiation, mitotic arrest and apoptosis of both imatinib-sensitive and imatinib-resistant CML cells. Finally, the anticancer activity of NCTU-322 against imatinib-resistant CML cells was demonstrated in vivo. In summary, our data demonstrated that andrographolide and NCTU-322 inhibit Bcr-abl function via a mechanism different from that of imatinib, and they induced multiple anticancer effects in both imatinib-sensitive and resistant CML cells. Our findings demonstrate that andrographolide and NCTU-322 are potential therapeutic agents again CML.Graphical abstractGraphical abstract for this article
  • Melatonin protects against methotrexate-induced memory deficit and
           hippocampal neurogenesis impairment in a rat model
    • Abstract: Publication date: Available online 23 February 2019Source: Biochemical PharmacologyAuthor(s): Apiwat Sirichoat, Suchada Krutsri, Kornrawee Suwannakot, Anusara Aranarochana, Pornthip Chaisawang, Wanassanun Pannangrong, Peter Wigmore, Jariya Umka Welbat Methotrexate (MTX) is a chemotherapy agent linked to cognitive deficits in cancer patients received chemotherapy treatment. MTX decreases cell proliferation in the hippocampus, which is concomitant with cognitive deficits in animal models. The present study aimed to investigate the disadvantages of MTX on cognition associated with cell division, survival, and immature neurons involved in hippocampal neurogenesis, as well as the practical neuroprotective effects of melatonin. Male Sprague Dawley rats were given two injections of MTX (75 mg/kg) on days 8 and 15 followed by Leucovorin (LCV, 6 mg/kg) at hours 18, 26, 42, 50 via i.p. injection. Some rats received co-treatment with melatonin (8 mg/kg, i.p. injection) for 15 days before and during MTX administration (preventive), 15 days after MTX administration (recovery), or both (30 days total; throughout). Hippocampal-dependent memory was examined using novel objection location (NOL) and novel object recognition (NOR) tests. Cell division, survival and immature neurons in the subgranular zone (SGZ) in the hippocampus were evaluated using immunofluorescence staining. Rats given MTX/LCV were found to have cognitive memory deterioration based on the NOL and NOR tests. Moreover, reductions in cell division, cell survival, and the numbers of immature neurons were detected in the MTX/LCV group when compared to the controls. This damage was not observed in rats in the preventive, recovery, or throughout groups. These findings reveal that melatonin has the potential to diminish the negative effects of MTX on memory and neurogenesis. This also indicates the benefit of melatonin co-administration in patients who undergo chemotherapy treatment.Graphical abstractGraphical abstract for this article
  • New options of cancer treatment employing InsP6
    • Abstract: Publication date: Available online 21 February 2019Source: Biochemical PharmacologyAuthor(s): Maria A. Brehm, Sabine Windhorst Many mechanistic studies have been performed to analyze the cellular functions of the highly phosphorylated molecule inositol hexakisphosphate (InsP6) in health and disease. While the physiological intracellular functions are well described, the mechanism of potential pharmacological effects on cancer cell proliferation is still controversial. There are numerous studies demonstrating that a high InsP6 concentration (≥75 µM) inhibits growth of cancer cells in vitro and in vivo. Thus, there is no doubt that InsP6 exhibits anticancer activity but the mechanism underlying the cellular effects of extracellular InsP6 on cancer cells is far from being understood. In addition, studies on the inhibitory effect of InsP6 on cancer progression in animal models ignore aspects of its bioavailability.Here, we review and critically discuss the uptake mechanism and the intracellular involvement in signaling pathways of InsP6 in cancer cells. We take into account the controversial findings on InsP6 plasma concentration, which is a critical aspect of pharmacological accessibility of InsP6 for cancer treatment. Further, we discuss novel findings with respect to the effect of InsP6 on normal and immune cells as well as on platelet aggregate size. Our goal is to stimulate further mechanistic studies into novel directions considering previously disregarded aspects of InsP6. Only when we fully understand the mechanism underlying the anticancer activity of InsP6 novel and more efficient treatment options can be developed.Graphical abstractGraphical abstract for this article
  • FCPR03, a novel phosphodiesterase 4 inhibitor, alleviates cerebral
           ischemia/reperfusion injury through activation of the AKT/GSK3β/
           β-catenin signaling pathway
    • Abstract: Publication date: Available online 21 February 2019Source: Biochemical PharmacologyAuthor(s): Bingtian Xu, Tiantian Wang, Jiao Xiao, Wenli Dong, Hui-zhen Wen, Xinyi Wang, Yunyun Qin, Ningbo Cai, Zhongzhen Zhou, Jiangping Xu, Haitao Wang Inhibition of phosphodiesterase 4 (PDE4) is a promising strategy for the treatment of ischemic stroke. However, the side effects of nausea and vomiting from the current PDE4 inhibitors have limited their clinical applications. FCPR03 is a novel PDE4 inhibitor with little emetic potential. This study aimed to investigate the effects of FCPR03 on neuronal injury after cerebral ischemia/reperfusion and the underlying signaling pathway. The effects of FCPR03 on cellular apoptosis, intracellular accumulation of reactive oxygen species (ROS), and mitochondrial membrane potential (MMP) were evaluated in HT-22 neuronal cells and cortical neurons exposed to oxygen-glucose deprivation (OGD). The impact of FCPR03 on brain injury, neurological scores and behavioral performance was investigated in rats subjected to middle cerebral artery occlusion (MCAO). The protein kinase B (AKT) inhibitor MK-2206 and β-catenin siRNA were used to investigate the underlying pathways. FCPR03 dose-dependently protected against OGD-induced cellular apoptosis in both HT-22 cells and cortical neurons. The levels of MMP and ROS were also restored by FCPR03. FCPR03 increased the levels of phosphorylated AKT, glycogen synthase kinase-3β (GSK3β), and β-catenin. Interestingly, the role of FCPR03 was reversed by MK-2206 and β-catenin siRNA. Consistently, FCPR03 reduced the infarct volume and improved neurobehavioral outcomes in rats following MCAO. Moreover, FCPR03 increased the levels of phosphorylated AKT, GSK3β and β-catenin within the ischemic penumbra of rats following cerebral ischemia-reperfusion. Taken together, FCPR03 has therapeutic potential in cerebral ischemia-reperfusion. The neuroprotective effects of FCPR03 are mediated through activation of the AKT/GSK3β/β-catenin pathway.Graphical abstractGraphical abstract for this article
  • Inhibition of mast cell degranulation by melanin
    • Abstract: Publication date: Available online 20 February 2019Source: Biochemical PharmacologyAuthor(s): Yoshiyuki Kawamoto, Hiromoto Kondo, Mari Hasegawa, Chiharu Kurimoto, Yuuki Ishii, Chihiro Kato, Taishi Botei, Muneshige Shinya, Takashi Murate, Yuki Ueno, Masao Kawabe, Yuko Goto, Ryohei Yamamoto, Machiko Iida, Ichiro Yajima, Nobutaka Ohgami, Masashi Kato, Kozue Takeda Melanin is a dark naturally occurring pigment produced in nature and in many organisms. Although several reports have demonstrated applications for melanins in various therapeutic treatments, to date, no research has examined the anti-allergic effect of melanin. In this study, we for the first time found that solubilized or synthesized soluble melanin acts as a potent inhibitor of the degranulation of mast cells. We found that squid-ink-derived melanin significantly inhibited antigen-IgE-FcεRI-mediated degranulation of the mucosal mast cell line RBL-2H3. A homogenized melanin nanoparticle prepared by laser ablation also clearly suppressed antigen-induced mast cell degranulation. We also successfully solubilized synthetic melanin in a neutral biochemical buffer and found that it also significantly inhibited IgE-sensitized mast cells. The anti-degranulation activity of synthesized melanin was abolished in the melanin fraction below 50-kD molecular weight. All melanins used in this study did not exert significant cell death. Signal transduction analysis revealed that melanin suppressed antigen-triggered phosphorylation of signaling molecules as well as calcium influx. Transmission electron microscopy revealed that homogenized melanin nanoparticles partially attached to the cell surface and some nanoparticles were internalized to the cell. Flow cytometry revealed that the number of FcεRI-bound IgE molecules was decreased by melanin. Fluorescence recovery after photobleaching analysis indicated that melanin attenuated both plasma membrane and cytoplasmic fluidity, implying that melanin increased their viscosities. In vivo experiments using passive systemic anaphylaxis (PSA) and passive cutaneous anaphylaxis (PCA) mouse models demonstrated that oral administration of melanin accelerated the recovery of decreased body temperature after antigen infection in PSA, and combination sensitization of IgE with melanin attenuated antigen-induced extravasation in PCA. These findings indicated that melanin exhibits preventative effects against IgE-mast cell-mediated anaphylaxis. This study provides the first evidence that homogenized melanin may be a potential therapeutic agent for diseases involving mast cells.Graphical abstractGraphical abstract for this article
  • Mechanisms of insulin resistance by simvastatin in C2C12 myotubes and in
           mouse skeletal muscle
    • Abstract: Publication date: Available online 20 February 2019Source: Biochemical PharmacologyAuthor(s): Gerda M. Sanvee, Miljenko V. Panajatovic, Jamal Bouitbir, Stephan Krähenbühl Statins inhibit cholesterol biosynthesis and lower serum LDL-cholesterol levels. They are generally well tolerated, but can cause insulin resistance in patients. Therefore, we investigated the mechanisms underlying the statin-induced insulin resistance.We used mice and C2C12 myotubes (murine cell line): mice (n=10) were treated with oral simvastatin (5 mg/kg/day) or water (control) for 21 days and C2C12 cells were exposed to 10 μM simvastatin for 24h.After intraperitoneal glucose application (2 g/kg), simvastatin-treated mice had higher glucose but equal insulin plasma concentrations than controls and lower glucose transport into skeletal muscle. Similarly, glucose uptake by C2C12 myotubes exposed to 10 μM simvastatin for 24h was impaired compared to control cells. In simvastatin-treated C2C12 myotubes, mRNA and protein expression of the insulin receptor (IR) β-chain was increased, but the phosphorylation (Tyr1361) was impaired. Simvastatin decreased numerically Akt/PKB Thr308 phosphorylation (via insulin signaling pathway) and significantly Akt/PKB Ser473 phosphorylation (via mTORC2), which was explained by impaired phosphorylation of mTOR Ser2448. Reduced phosphorylation of Akt/PKB impaired downstream phosphorylation of GSK3β, leading to impaired translocation of GLUT4 into plasma membranes of C2C12 myotubes. In contrast, reduced phosphorylation of AS160 could be excluded as a reason for impaired GLUT4 translocation.In conclusion, simvastatin caused insulin resistance in mice and impaired glucose uptake in C2C12 myotubes. The findings in myotubes can be explained by diminished activation of Akt/PKB by mTORC2 and downstream effects on GSK3β, impairing the translocation of GLUT4 and the uptake of glucose.Graphical abstractGraphical abstract for this article
  • The transcription factor E4bp4 regulates the expression and activity of
           Cyp3a11 in mice
    • Abstract: Publication date: Available online 20 February 2019Source: Biochemical PharmacologyAuthor(s): Yongbin Tong, Peng Zeng, Tianpeng Zhang, Mengjing Zhao, Pei Yu, Baojian Wu Human CYP3A4 (Cyp3a11 in mice) is one of the most important enzymes for drug metabolism and detoxification. Here, we aimed to investigate a potential role for E4bp4 in regulation of Cyp3a11 expression and activity. The regulatory effects of E4bp4 on Cyp3a11 enzyme were assessed using E4bp4-/- mice and Hepa-1c1c7 cells. The mRNA and protein levels were quantified using qPCR and Western blotting, respectively. In vitro microsomal Cyp3a11 activity was probed using its specific substrates midazolam and testosterone. Pharmacokinetic studies were performed with wild-type and E4bp4-/- mice after midazolam administration. Global deletion of E4bp4 led to significant upregulation of Cyp3a11 mRNA and protein in major metabolic organs (i.e., the liver, kidney and small intestine). E4bp4 ablation also caused an increased microsomal Cyp3a11 activity consistent with the enzyme’s expression change. Overexpression of E4bp4 in Hepa-1c1c7 cells resulted in reduced levels of Cyp3a11 mRNA and protein, whereas E4bp4 knockdown caused upregulation of Cyp3a11 expression. In addition, the systemic exposure of midazolam was lowered in E4bp4-/- mice compared with wild-type mice. This was accompanied by enhanced formation of its metabolite 1’-hydroxymidazolam. Furthermore, luciferase reporter and mobility shift assays revealed that E4bp4 repressed Cyp3a11 transcription via direct binding to C-site (-1539/-1529 bp) in the promoter region. In conclusion, E4bp4 negatively regulates Cyp3a11 expression, thereby impacting drug metabolism and pharmacokinetics.Graphical abstractGraphical abstract for this article
  • Aryl hydrocarbon receptor (AHR) functions in NAD+ metabolism,
           myelopoiesis and obesity
    • Abstract: Publication date: Available online 16 February 2019Source: Biochemical PharmacologyAuthor(s): Karl Walter Bock Diverse physiologic functions of AHR, a transcription factor discovered in studies of dioxin toxicity, are currently elucidated in many laboratories including chemical and microbial defense, immunity and myelopoiesis. Accumulating evidence suggests that AHR may also be involved in obesity and TCDD-mediated lethality in sensitive species. Underlying mechanisms include NAD+- and sirtuin-mediated deregulation of lipid, glucose and NAD+ homeostasis. Progress in NAD metabolome research suggests large consumption of NAD+ by NAD glycohydrolases (NADases) and NAD-dependent sirtuins. In focus are two NADases: (i) TiPARP (TCDD-induced poly(ADP-ribose) polymerase), one of several nuclear NADases, and (ii) plasma membrane-bound ectoNADase/CD38, a multifunctional enzyme and receptor. CD38 is involved in extra- and intracellular NAD degradation but acts also as differentiation marker. Both CD38 and AHR are components of a complex signalsome that enhances retinoic acid-induced differentiation of myeloid progenitor cells to granulocytes. Further advances of NAD metabolome research may lead to therapeutic options in the control of obesity and to improved risk assessment of TCDD toxicity.Graphical abstractGraphical abstract for this article
  • Ca2+/calmodulin-dependent protein kinase II regulation by inhibitor 1 of
           protein phosphatase 1 alleviates necroptosis in high glucose-induced
           cardiomyocytes injury
    • Abstract: Publication date: Available online 16 February 2019Source: Biochemical PharmacologyAuthor(s): Linlin Sun, Yun Chen, Huiqin Luo, Mengting Xu, Guoliang Meng, Wei Zhang Ca2+/calmodulin-dependent protein kinase II (CaMKII) plays an important role in the cardiovascular system. However, the potential protective role of inhibitor 1 of protein phosphatase 1 (I1PP1), which is able to regulate CaMKII, in high glucose-induced cardiomyocytes injury remains unknown. In the present study, cardiomyocytes were transfected with I1PP1 adenovirus to inhibit protein phosphatase 1 (PP1) expression. After the cardiomyocytes were subjected to high glucose stimulation for 48 h, quantitative real-time PCR was used to detect CaMKIIδ alternative splicing. Lactate dehydrogenase (LDH) release and adenosine triphosphate (ATP) level were measured to assess cell damage and energy metabolism respectively. CaMKII activity was represented as phospholamban (PLB) phosphorylation, CaMKII phosphorylation (p-CaMKII) and oxidation (ox-CaMKII). Dihydroethidium (DHE), MitoSOX and JC-1 staining were used to assess oxidative stress and mitochondrial membrane potential. Necroptosis was evaluated by receptor interacting protein kinase 3 (RIPK3) expression, TUNEL and cleaved-caspase 3 levels. RIPK3, mixed lineage kinase domain like protein (MLKL) and dynamin-related protein 1 (DRP1) expressions were also detected. We found that high glucose disordered CaMKIIδ alternative splicing. I1PP1 over-expression suppressed PLB phosphorylation, ox-CaMKII, DRP1, RIPK3 and cleaved-caspase 3 proteins expression, decreased LDH release, attenuated necroptosis, increased ATP level, inhibited oxidative stress, and elevated mitochondrial membrane potential in high glucose-stimulated cardiomyocytes. However, there was no effect on phosphorylation of MLKL (p-MLKL), p-CaMKII, and receptor interacting protein kinase (RIPK1) expression. Altogether, I1PP1 over-expression alleviated CaMKIIδ alternative splicing disorder, suppressed CaMKII oxidation, reduced reactive oxygen species (ROS) accumulation and inhibited necroptosis to attenuate high glucose-induced cardiomyocytes injury.Graphical abstractGraphical abstract for this article
  • Nox2-mediated platelet activation by glycoprotein (GP) VI: Effect of
           rivaroxaban alone and in combination with aspirin
    • Abstract: Publication date: May 2019Source: Biochemical Pharmacology, Volume 163Author(s): Vittoria Cammisotto, Roberto Carnevale, Cristina Nocella, Lucia Stefanini, Simona Bartimoccia, Antonio Coluccia, Romano Silvestri, Pasquale Pignatelli, Daniele Pastori, Francesco Violi Factor Xa (FXa) has been reported to activate platelet via interaction with glycoprotein (GP) VI but the underlying mechanism has not been fully elucidated. We investigated if Nox2-derived oxidative stress is implicated in FXa-induced platelet aggregation (PA), and the effect of a FXa inhibitor, namely rivaroxaban, with or without aspirin (ASA), on PA.We performed an in vitro study measuring convulxin-induced PA, thromboxane (Tx) B2 and isoprostanes biosynthesis, soluble Nox2-dp (sNox2-dp), a marker of Nox2 activation, soluble GPVI (sGPVI) and PLA2 activation in platelets from healthy subjects (n = 5) added with and without a Nox2 inhibitor. The same variables were also examined in platelets treated with rivaroxaban (15–60 ng/ml), combined or less with ASA (25 µM).Convulxin-stimulated platelets increased sGPVI, sNox2-dp, H2O2, eicosanoid biosynthesis and PLA2 phosphorylation, which were all inhibited by a Nox2 inhibitor. Rivaroxaban alone significantly reduced PA, sGPVI, TxB2 and isoprostanes biosynthesis, concomitantly with Syk, sNox2-dp and PLA2 activation in a dose-dependent fashion; a significant effect was achieved with 30 ng/ml rivaroxaban. Docking simulation analysis showed that rivaroxaban interacts with GPVI. In platelets co-incubated with ASA, rivaroxaban amplified the ASA antiplatelet effect, which was achieved with 30 ng/ml and prevalently attributable to Nox2 inhibition and impaired isoprostane biosynthesis.Here we show that rivaroxaban, at concentrations achievable in human circulation, inhibits PA via GPVI interaction and eventually Nox2-mediated isoprostanes biosynthesis and amplifies the ASA antiplatelet effect.Graphical abstractRed and blue arrows show the rivaroxaban and convulxin (CVX) pathways respectively. Rivaroxaban binds the GPVI receptor, inhibits Nox2-derived oxidative stress and ultimately platelet Thromboxane A2 and isoprostanes production.Graphical abstract for this article
  • Glycogen synthase kinase-3 inhibitor as a multi-targeting anti-rheumatoid
    • Abstract: Publication date: Available online 15 February 2019Source: Biochemical PharmacologyAuthor(s): Masaki Arioka, Fumi Takahashi-Yanaga Rheumatoid arthritis (RA) is a chronic inflammatory joint disease that causes swelling, bone erosion, and joint disorder. Patients with RA therefore suffer from pain and physiological disability, and have a decreased quality of life. During the progression of RA, many different types of cells and inflammatory factors influence each other with an important role. A better understanding of the pathology of RA should therefore lead to the development of effective anti-rheumatoid drugs, such as the anti-TNFα antibody. Glycogen synthase kinase-3 (GSK-3) is a cytoplasmic serine/threonine protein kinase that is involved in a large number of key cellular processes and is dysregulated in a wide variety of diseases, including inflammation and osteoporosis. The accumulated evidence has suggested that GSK-3 could be involved in multiple steps in the progression of RA. In the present review, the mechanisms of the pathogenesis of RA are summarized, and recent developments and potential new drugs targeting GSK-3 are discussed.Graphical abstractGraphical abstract for this article
  • Functional characterization of AC5 gain-of-function variants: Impact on
           the molecular basis of ADCY5-related dyskinesia
    • Abstract: Publication date: Available online 14 February 2019Source: Biochemical PharmacologyAuthor(s): T.B. Doyle, M.P. Hayes, D.H. Chen, W.H. Raskind, V.J. Watts Adenylyl cyclases are key points for the integration of stimulatory and inhibitory G protein-coupled receptor (GPCR) signals. Adenylyl cyclase type 5 (AC5) is highly expressed in striatal medium spiny neurons (MSNs), and is known to play an important role in mediating striatal dopaminergic signaling. Dopaminergic signaling from the D1 expressing MSNs of the direct pathway, as well as the D2 expressing MSNs of the indirect pathway both function through the regulation of AC5 activity, controlling the production of the 2nd messenger cAMP, and subsequently the downstream effectors. Here, we used a newly developed cell line that used Crispr-Cas9 to eliminate the predominant adenylyl cyclase isoforms to more accurately characterize a series of AC5 gain-of-function mutations which have been identified in ADCY5-related dyskinesias. Our results demonstrate that these AC5 mutants exhibit enhanced activity to Gαs-mediated stimulation in both cell and membrane-based assays. We further show that the increased cAMP response at the membrane effectively translates into increased downstream gene transcription in a neuronal model. Subsequent analysis of inhibitory pathways show that the AC5 mutants exhibit significantly reduced inhibition following D2 dopamine receptor activation. Finally, we demonstrate that an adenylyl cyclase “P-site” inhibitor, SQ22536 may represent an effective future therapeutic mechanism by preferentially inhibiting the overactive AC5 gain-of-function mutants.Graphical abstractGraphical abstract for this article
  • The novel hybrid agonist HyNDA-1 targets the D3R-nAChR heteromeric complex
           in dopaminergic neurons
    • Abstract: Publication date: Available online 14 February 2019Source: Biochemical PharmacologyAuthor(s): Carlo Matera, Federica Bono, Silvia Pelucchi, Ginetta Collo, Leonardo Bontempi, Cecilia Gotti, Michele Zoli, Marco De Amici, Cristina Missale, Chiara Fiorentini, Clelia Dallanoce In this paper, we designed, synthesized and tested a small set of three new derivatives potentially targeting the D3R-nAChR heteromer, a receptor complex recently identified and characterized as the molecular entity that, in dopaminergic neurons, mediates the neurotrophic effects of nicotine. By means of a partially rigidified spacer of variable length, we incorporated in the new compounds (1a-c) the pharmacophoric substructure of a known β2-subunit-containing nAChR agonist (A-84543) and that of the D2/D3R agonist drug ropinirole. All the compounds retained the ability to bind with high affinity both β2-subunit-containing nAChR and D3R. Compound 1a, renamed HyNDA-1, which is characterized by the shortest linker moiety, was the most interesting ligand. We found, in fact, that HyNDA-1 significantly modulated structural plasticity on both mice and human dopaminergic neurons, an effect strongly prevented by co-incubating this ligand with either nAChR or D3R antagonists. Moreover, the neurotrophic effects of HyNDA-1 were specifically lost by disrupting the complex with specific interfering peptides. Interestingly, by using the Bioluminescence Resonance Energy Transfer 2 (BRET2) assay in HEK-293 transfected cells, we also found that HyNDA-1 has the ability to increase the affinity of interaction between nAChR and D3R. Overall, our results indicate that the neurotrophic effects of HyNDA-1 are mediated by activation of the D3R-nAChR heteromeric complex specifically expressed on dopaminergic neurons.Graphical abstractGraphical abstract for this article
  • The role of the Hint1 protein in the metabolism of phosphorothioate
           oligonucleotides drugs and prodrugs, and the release of H2S under cellular
    • Abstract: Publication date: Available online 14 February 2019Source: Biochemical PharmacologyAuthor(s): Krakowiak Agnieszka, Piotrzkowska Danuta, Kocoń-Rębowska Beata, Kaczmarek Renata, Maciaszek Anna Phosphorothioate oligonucleotides (PS-oligos) containing sulfur atom attached in a nonbridging position to the phosphorus atom at one or more internucleotide bond(s) are often used in medicinal applications. Their hydrolysis in cellular media proceeds mainly from the 3’-end, resulting in the appearance of nucleoside 5’-O-phosphorothioates ((d)NMPS), whose further metabolism is poorly understood. We hypothesize that the enzyme responsible for (d)NMPS catabolism could be Hint1, an enzyme that belongs to the histidine triad (HIT) superfamily and is present in all organisms. We previously found that (d)NMPS were desulfurated in vitro to yield (d)NMP and H2S in a Hint1-assisted reaction. Here, we demonstrate that AMPS/GMPS/dGMPS introduced into HeLa/A549 cells are intracellularly converted into AMP/GMP/dGMP and H2S. The level of the released H2S was relative to the concentration of the compounds used and the reaction time. Using RNAi technology, we have shown decreased levels of AMPS/GMPS desulfuration in HeLa/A549 cells with reduced Hint1 levels. Finally, after transfection of a short Rp-d(APSAPSA) oligomer into HeLa cells, the release of H2S was observed. These results suggest that the metabolic pathway of PS-oligos includes hydrolysis into (d)NMPS (by cellular nucleases) followed by Hint1-promoted conversion of the resulting (d)NMPS into (d)NMP accompanied by H2S elimination. Our observations may be also important for possible medicinal applications of (d)NMPS because H2S is a gasotransmitter involved in many physiological and pathological processes.Graphical abstractGraphical abstract for this article
  • Palbociclib triggers apoptosis in bladder cancer cells by Cdk2-induced
           Rad9-mediated reorganization of the Bak.Bcl-xl complex
    • Abstract: Publication date: Available online 14 February 2019Source: Biochemical PharmacologyAuthor(s): Guohai Zhang, Feng'e Ma, Liangping Li, Jingjing Li, Pingping Li, Shulan Zeng, Hongbin Sun, Erguang Li Palbociclib is a Cdk4/6 inhibitor approved for metastatic estrogen receptor-positive breast cancer. The drug is also under clinical evaluation for metastatic urothelial cancer and other solid tumors. Preclinical studies from multiple tumor types suggest that other factors also affect the sensitivity of individual tumors to Cdk4/6 inhibitor. We show here that Cdk2 has an essential role in palbociclib antitumor effect against bladder cancers. We found that palbociclib induced apoptosis instead of cell cycle arrest to exhibit its anticancer activity in T24 cells, as was evidenced by membrane blebbing, caspase-3 activation and AIF release from mitochondria. Cdk2 activation was important to palbociclib-induced apoptotic triggering activity, since depletion of Cdk2 significantly inhibited caspase-3 activation and cell apoptosis. Cdk2 activation caused p-Rad9 translocation to the mitochondria and subsequently interaction with Bcl-xl, leading to conformational activation of Bak and cell apoptosis. The anticancer activity and Cdk2 activation of palbociclib-treated mice were finally validated in a T24 xenograft model. Collectively, these results together demonstrate that palbociclib exerts its anticancer effect in T24 cells mainly through Cdk2 activation. Our findings provide new insights into the molecular interactions and anticancer mechanisms of Cdk4/6 inhibitors.Graphical abstractGraphical abstract for this article
  • Characterization of aspirin esterase activity in health and disease: in
           vitro and ex vivo studies
    • Abstract: Publication date: Available online 13 February 2019Source: Biochemical PharmacologyAuthor(s): Benedetta Porro, Alessandro Di Minno, Bianca Rocca, Susanna Fiorelli, Sonia Eligini, Linda Turnu, Simone Barbieri, Alessandro Parolari, Elena Tremoli, Viviana Cavalca Due to its ability to irreversibly inactivate platelet cyclooxygenase, low-dose aspirin (ASA) is the most widely used antithrombotic agent. Although, studies in specific types of patients with cardiovascular disease (CVD) have shown an incomplete inhibition of platelet’s cyclooxygenase, which may increase the variability in drug response. Some aspects of ASA pharmacokinetics (PK) still need further investigation. In this study, we aimed to characterise the contribution of esterase enzymes to ASA hydrolysis in the peripheral blood and to assess their activity in 36 healthy subjects (Ctrl) and 77 CVD patients. To this aim, an in vitro assay testing esterase activity in parallel to a liquid chromatography-tandem mass spectrometry method simultaneously detecting ASA and its main metabolites salicylic (SA) and gentisic acids, have been developed. Michaelis-Menten constant (Km) calculation, ASA esterase isoforms characterisation, and ASA PK study were then achieved. The calculated Km identified plasma esterases as the enzymes with the higher affinity for the substrate compared to the RBC ones. Both rivastigmine and 4-bis-nitrophenyl phosphate inhibited plasma esterase activities, suggesting that acetylcholinesterase and carboxylesterase largely contribute to ASA hydrolysis. The feasibility of the method here developed has been explored in Ctrl and CVD patients. The effect of ASA treatment on enzyme activity was further evaluated on an age, sex and BMI matched subgroup of patients and Ctrl (n=10 for each subgroup, on and off ASA). No overall variations were evidenced in both CVD and Ctrl groups, even when the effect of ASA treatment was tested. This result suggests the absence of any influence of disease state, drug treatments, and comorbidities on plasma esterase and the inability of ASA intake to induce esterase function. In conclusion, the method here developed allows a better characterisation of ASA esterase activity and could be helpful to define the PK-related determinants of ASA responsiveness in order to personalise regimen in specific pathological conditions.Graphical abstractGraphical abstract for this article
  • Alkylphenol inverse agonists of HCN1 gating: H-bond propensity, ring
           saturation and adduct geometry differentially determine efficacy and
    • Abstract: Publication date: Available online 13 February 2019Source: Biochemical PharmacologyAuthor(s): Rebecca L Joyceadat, Nicole P Beyer, Georgia Vasilopoulos, Kellie Woll, Adam C Hall, Roderic G Eckenhoff, Dipti N Barman, J David Warren, Gareth R Tibbs, Peter A Goldstein Background and purposeIn models of neuropathic pain, inhibition of HCN1 is anti-hyperalgesic. 2,6-di-iso-propyl phenol (propofol) and its non-anesthetic congener, 2,6-di-tert-butyl phenol, inhibit HCN1 channels by stabilizing closed state(s).ExperimentalapproachUsing in vitro electrophysiology and kinetic modeling, we systematically explore the contribution of ligand architecture to alkylphenol-channel coupling.Key resultsWhen corrected for changes in hydrophobicity (and propensity for intra-membrane partitioning), the decrease in potency upon 1-position substitution (NCO∼OH>> SH>>> F) mirrors the ligands’ H-bond acceptor (NCO> OH> SH>>> F) but not donor profile (OH> SH>>> NCO∼F). H-bond elimination (OH to F) corresponds to a ΔΔG of ∼4.5 kCal mol-1 loss of potency with little or no disruption of efficacy. Substitution of compact alkyl groups (iso-propyl, tert-butyl) with shorter (ethyl, methyl) or more extended (sec-butyl) adducts disrupts both potency and efficacy. Ring saturation (with the obligate loss of both planarity and π electrons) primarily disrupts efficacy.Conclusions and implicationsA hydrophobicity-independent decrement in potency at higher volumes suggests the alkylbenzene site has a volume of ≥ 800 Å3. Within this, a relatively static (with respect to ligand) H-bond donor contributes to initial binding with little involvement in generation of coupling energy. The influence of π electrons/ring planarity and alkyl adducts on efficacy reveals these aspects of the ligand present towards a face of the channel that undergoes structural changes during opening. The site’s characteristics suggest it is “druggable”; introduction of other adducts on the ring may generate higher potency inverse agonists.Graphical abstractGraphical abstract for this article
  • Impairment of chemical hypoxia-induced sphingosine kinase-1 expression and
           activation in rheumatoid arthritis synovial fibroblasts: A signature of
    • Abstract: Publication date: Available online 10 February 2019Source: Biochemical PharmacologyAuthor(s): Chenqi Zhao, Nathalie Amiable, Melissa Laflamme, David Marsolais, John A. Di Battista, Maria J. Fernandes, Sylvain G. Bourgoin Sphingosine kinase 1 (SphK1) and 2 (SphK2) have been shown contribute to synovial inflammation in animal models of arthritis. However, low levels of intracellular sphingosine-1 phosphate (S1P) were reported in fibroblast-like synoviocytes (FLS) from patients in the end stage of rheumatoid arthritis (RA) compared to normal FLS. Moreover, the S1P receptor-mediated chemokine synthesis was altered in RAFLS in response to chemical hypoxia. Since the mechanisms responsible for low levels of intracellular S1P in RAFLS are not fully identified, we evaluated the contribution of SphKs to the S1P-induced synthesis of chemokines under conditions of chemical hypoxia. Our results show that a chemical hypoxia mimetic cobalt chloride (CoCl2) increased SphK1 expression and activation in normal FLS but not in RAFLS. Using selective inhibitors of SphKs and gene silencing approaches, we provide evidence that both SphK1 and SphK2 are involved in hypoxia-induced chemokine production in normal FLS. In contrast, only SphK2 mediates hypoxia-induced chemokine production in RAFLS. Moreover, CoCl2 increased S1P2 and S1P3 receptor mRNA levels in normal FLS but not in RAFLS. The data suggest that altered expression and/or activation of SphK1 combined with reduced induction of S1P receptor expression by CoCl2 impaired the CoCl2-mediated autocrine S1P receptor signaling loop and chemokine production in RAFLS.Graphical abstractSphK1 expression and/or activation are impaired in hypoxic RAFLS, leading to reduced autocrine S1PR signalling/chemokine loop. (↓: decrease, ↑: increase)Graphical abstract for this article
  • A novel soluble guanylyl cyclase activator, BR 11257, acts as a
           non-stabilising partial agonist of sGC
    • Abstract: Publication date: Available online 10 February 2019Source: Biochemical PharmacologyAuthor(s): Christin Elgert, Anne Rühle, Peter Sandner, Sönke Behrends The soluble guanylyl cyclase (sGC) plays a key role in NO/cGMP signalling and is widely recognised to be important in different disease pathomechanisms. The discovery of sGC agonists provides a new opportunity to stimulate the NO/cGMP pathway. One class of compounds are the heme-independent sGC activators, which are thought to bind to oxidised or heme-free sGC. This enzyme is preferentially formed under disease situations accompanied by oxidative stress. Accordingly, this binding mode of sGC activators has quite some appeal for the clinical use of sGC activator drugs in diseases with high oxidative stress burden. However, none of the previous sGC activators, most of them dicarboxylic acid derivatives, has passed clinical trials to date, also because of the potent blood pressure lowering effects. In the current study, we investigate the effects of a new monocarboxylic drug BR 11257 in vitro and in vivo. Activity measurements with purified enzyme indicated gentle sGC activation for BR 11257 resembling a partial agonistic behaviour. In thermal shift measurements, we observed an unexpected difference between BR 11257 and the sGC activators from the dicarboxylic acid type. While activators from the dicarboxylic acid type had a highly thermostabilising influence on sGC, this effect was absent with BR 11257. We hypothesize that the key interaction partner for thermostabilisation is the second carboxylic acid in BAY 60-2770 which is missing in BR 11257. The absence of this thermodynamic receptor stabilisation and the partial agonism may be advantageous to overcome limitations of this class of drugs by avoiding excessive hypotension.Graphical abstractGraphical abstract for this article
  • Piperlongumine-induced nuclear translocation of the FOXO3A transcription
           factor triggers BIM-mediated apoptosis in cancer cells
    • Abstract: Publication date: Available online 10 February 2019Source: Biochemical PharmacologyAuthor(s): Zhenxing Liu, Zhichen Shi, Jieru Lin, Shuang Zhao, Min Hao, Junting Xu, Yuyin Li, Qing Zhao, Li Tao, Aipo Diao The transcription factor forkhead box O 3A (FOXO3A) is a tumor suppressor that promotes cell cycle arrest and apoptosis. Piperlongumine (PL), a plant alkaloid, is known to selectively kill tumor cells while sparing normal cells. However, the mechanism of PL-induced cancer cell death is not fully understood. We report here that an association of FOXO3A with the pro-apoptotic protein BIM (also known as BCL2-like 11, BCL2L11) has a direct and specific function in PL-induced cancer cell death. Using HeLa cells stably expressing a FOXO3A-GFP fusion protein and several other cancer cell lines, we found that PL treatment induces FOXO3A dephosphorylation and nuclear translocation and promotes its binding to the BIM gene promoter, resulting in the up-regulation of BIM in the cancer cell lines. Accordingly, PL inhibited cell viability and caused intrinsic apoptosis in a FOXO3A-dependent manner. Of note, siRNA-mediated FOXO3A knockdown rescued the cells from PL-induced cell death. In vivo, the PL treatment markedly inhibited xenograft tumor growth, and this inhibition was accompanied by the activation of the FOXO3A-BIM axis. Moreover, PL promoted FOXO3A dephosphorylation by inhibiting phosphorylation and activation of Akt, a kinase that phosphorylates FOXO3A. In summary, our findings indicate that PL activates the FOXO3A-BIM apoptotic axis by promoting dephosphorylation and nuclear translocation of FOXO3A via Akt signaling inhibition. These findings uncover a critical mechanism underlying the effects of PL on cancer cells.Graphical abstractGraphical abstract for this article
  • Time-dependent course of gastric ulcer healing and molecular markers
           profile modulated by increased gastric mucosal content of carbon monoxide
           released from its pharmacological donor
    • Abstract: Publication date: Available online 10 February 2019Source: Biochemical PharmacologyAuthor(s): Katarzyna Magierowska, Dominik Bakalarz, Dagmara Wójcik, Anna Chmura, Magdalena Hubalewska-Mazgaj, Sabina Licholai, Edyta Korbut, Slawomir Kwiecien, Zbigniew Sliwowski, Grzegorz Ginter, Tomasz Brzozowski, Marcin Magierowski Background and PurposeBesides hydrogen sulfide (H2S) and nitric oxide (NO), carbon monoxide (CO) contributes to the maintenance of gastric mucosal integrity. We investigated increased CO bioavailability effects on time-dependent dynamics of gastric ulcer healing mediated by particular growth factors, anti-inflammatory and molecular pathways.Experimental ApproachWistar rats with gastric ulcers induced by serosal acetic acid application (day 0) were treated i.g. throughout 3, 6 or 14 days with vehicle or CO-releasing tricarbonyldichlororuthenium (II) dimer (CORM-2, 2.5 mg/kg). Gross and microscopic alterations in gastric ulcer size and gastric blood flow (GBF) at ulcer margin were determined by planimetry, histology and laser flowmetry, respectively. Gastric mRNA/protein expressions of platelet derived growth factors (PDGFA-D), insulin-like growth factor (IGF-1), epidermal growth factor (EGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGFA) and their receptors, heme oxygenases (HMOX), nuclear factor (erythroid-derived 2)-like 2 (Nrf-2), cyclooxygenase (COX-2), hypoxia inducible factor (HIF)-1α, anti-inflammatory annexin-1 and transforming growth factor (TGF-β1) were assessed by real-time PCR or Western blot. TGF-β1-3 and IL-10 plasma concentration were measured using Luminex platform. Prostaglandin E2 content at ulcer margin was assessed by ELISA.KeyresultsCORM-2 decreased ulcer area and increased GBF after 6 and 14 days comparing to vehicle. CO donor upregulated HGF, HGFr, VEGFR1, VEGFR2, TGF-β1, annexin-1 and maintained increased IGF-1, PDGFC and EGF expression at time-intervals of ulcer healing. TGF-β3 and IL-10 plasma concentration were significantly increased after COMR-2 vs. vehicle.ConclusionsCO time-dependently accelerates gastric ulcer healing and raises GBF at ulcer margin by mechanism involving subsequent upregulation of anti-inflammatory, growth promoting and angiogenic factors response, not observed physiologically.Graphical abstractGraphical abstract for this article
  • Small molecule nicotinamide N-methyltransferase inhibitor activates
           senescent muscle stem cells and improves regenerative capacity of aged
           skeletal muscle
    • Abstract: Publication date: Available online 10 February 2019Source: Biochemical PharmacologyAuthor(s): Harshini Neelakantan, Camille R. Brightwell, Ted Gfoch. Graber, Rosario Maroto, Hua-Yu Leo Wang, Stanton F. McHardy, John Papaconstantinou, Christopher S. Fry, Stanley J. Watowich Aging is accompanied by progressive declines in skeletal muscle mass and strength and impaired regenerative capacity, predisposing older adults to debilitating age-related muscle deteriorations and severe morbidity. Muscle stem cells (muSCs) that proliferate, differentiate to fusion-competent myoblasts, and facilitate muscle regeneration are increasingly dysfunctional upon aging, impairing muscle recovery after injury. While regulators of muSC activity can offer novel therapeutics to improve recovery and reduce morbidity among aged adults, there are no known muSC regenerative small molecule therapeutics. We recently developed small molecule inhibitors of nicotinamide N-methyltransferase (NNMT), an enzyme overexpressed with aging in skeletal muscles and linked to impairment of the NAD+ salvage pathway, dysregulated sirtuin 1 activity, and increased muSC senescence. We hypothesized that NNMT inhibitor (NNMTi) treatment will rescue age-related deficits in muSC activity to promote superior regeneration post-injury in aging muscle. 24-month old mice were treated with saline (control), and low and high dose NNMTi (5 and 10 mg/kg) for 1-week post-injury, or control and high dose NNMTi for 3-weeks post-injury. All mice underwent an acute muscle injury (barium chloride injection) locally to the tibialis anterior (TA) muscle, and received 5-ethynyl-2’-deoxyuridine systemically to analyze muSC activity. In vivo contractile function measurements were conducted on the injured TA muscle and tissues collected for ex-vivo analyses, including myofiber cross-sectional area (CSA) measurements to assess muscle recovery. Results revealed that muscle stem cell proliferation and subsequent fusion were elevated in NNMTi-treated mice, supporting nearly 2-fold greater CSA and shifts in fiber size distribution to greater proportions of larger sized myofibers and fewer smaller sized fibers in NNMTi-treated mice compared to controls. Prolonged NNMTi treatment post-injury further augmented myofiber regeneration evinced by increasingly larger fiber CSA. Importantly, improved muSC activity translated not only to larger myofibers after injury but also to greater contractile function, with the peak torque of the TA increased by ∼70% in NNMTi-treated mice compared to controls. Similar results were recapitulated in vitro with C2C12 myoblasts, where NNMTi treatment promoted and enhanced myoblast differentiation with supporting changes in the cellular NAD+/NADH redox states. Taken together, these results provide the first clear evidence that NNMT inhibitors constitute a viable pharmacological approach to enhance aged muscle regeneration by rescuing muSC function, supporting the development of NNMTi as novel mechanism-of-action therapeutic to improve skeletal muscle regenerative capacity and functional recovery after musculoskeletal injury in older adults.Graphical abstractGraphical abstract for this article
  • AMPK: a promising molecular target for combating cisplatin toxicities
    • Abstract: Publication date: Available online 7 February 2019Source: Biochemical PharmacologyAuthor(s): Nadereh Rashtchizadeh, Hassan Argani, Amir Ghorbani haghjo, Davoud Sanajou, Vahid Hosseini, Siavoush Dastmalchi, Saeed Nazari Soltan Ahmad Cisplatin is a broadly prescribed anti-tumor agent for the treatment of diverse cancers. Therapy with cisplatin, however, is associated with various adverse effects including nephrotoxicity and ototoxicity. AMP kinase (AMPK), an evolutionarily conserved enzyme, functions as the fundamental regulator of energy homeostasis. While AMPK activation protects normal tissues against cisplatin-induced toxicities, its impact in cancer is context-dependent and there is no single, uniform role for AMPK. On one hand, some report that AMPK activation augments cisplatin-induced apoptosis in cancer, while on the other hand, few reports indicate that AMPK activation rescues cancer cells from the cytotoxicity induced by cisplatin. Here we review the most salient signaling pathways regulated by AMPK with an emphasis on their relation to cisplatin toxicity and yet discuss context-dependent functions of AMPK in cancer.Graphical abstractGraphical abstract for this article
  • 7,8-dihydroxyflavone blocks the development of behavioral sensitization to
           MDPV, but not to cocaine: differential role of the BDNF-TrkB pathway
    • Abstract: Publication date: Available online 6 February 2019Source: Biochemical PharmacologyAuthor(s): L. Duart-Castells, R. López-Arnau, S. Vizcaíno, J. Camarasa, D. Pubill, E. Escubedo 3,4-Methylenedioxypyrovalerone (MDPV) acts as a dopamine transporter blocker and exerts powerful psychostimulant effects. In this study we aimed to investigate the bidirectional cross-sensitization between MDPV and cocaine, as well as to evaluate the role of the BDNF-TrkB signaling pathway in the development of locomotor sensitization to both drugs.Mice were treated with MDPV (1.5 mg/kg) or cocaine (10 or 15 mg/kg) once daily for 5 days. After withdrawal (10 days), animals were challenged with cocaine (8 mg/kg) or MDPV (1 mg/kg). For biochemical determinations, MDPV (1.5 mg/kg) or cocaine (15 mg/kg) were administered acutely or repeatedly, and BDNF, D3R and G9a transcription levels as well as pro- and mature BDNF protein levels were determined.Our results demonstrate that repeated administration of MDPV or cocaine sensitizes to cocaine and MDPV locomotor effects. After an acute or a repeated exposure to MDPV, cortical mRNA BDNF levels were increased, while a decrease in mBDNF protein levels in the nucleus accumbens 2h after repeated exposure was evidenced. Interestingly, such decline was involved in the development of locomotor sensitization, thus the pretreatment with 7,8-dihydroxyflavone (10 mg/kg), a TrkB agonist, blocked the development of sensitization to MDPV but not to cocaine, for which no changes in the BDNF-TrkB signaling pathway were observed at early withdrawal.In conclusion, a bidirectional cross-sensitization between MDPV and cocaine was evidenced. Our findings suggest that decreased BDNF-TrkB signaling has an important role in the behavioral sensitization to MDPV, pointing TrkB modulation as a target to prevent MDPV sensitization.Graphical abstractGraphical abstract for this article
  • Helicobacter pylori infection promotes autophagy through Nrf2-mediated
           heme oxygenase upregulation in human gastric cancer cells
    • Abstract: Publication date: Available online 4 February 2019Source: Biochemical PharmacologyAuthor(s): Ji Yeon Paik, Hee Geum Lee, Juan-Yu Piao, Su-Jung Kim, Do-Hee Kim, Hye-Kyung Na, Young-Joon Surh It has been reported that Helicobacter pylori (H. pylori) infection is one of the primary causes of gastritis and peptic ulcer diseases. More than 50% of the world’s population is supposed to be infected by this bacterium. However, 90% of infected patients are asymptomatic, suggesting the existence of host defense mechanisms. Nrf2 is a transcription factors that plays a key role in cellular defence against oxidative stress and inflammation. Autophagy, an autodigestive process that degrades cellular organelles and proteins, plays an important role in maintaining cellular homeostasis. To investigate the molecular mechanisms responsible for cellular adaptive response to H. pylori induced gastric inflammation, human gastric epithelial cells and mice were infected with H. pylori. H. pylori infection induced expression of microtubule-associated light chain3 (LC3), an autophagic marker, through accumulation of reactive oxygen species and subsequently Nrf2 nuclear translocation in AGS cells. Furthermore, Nrf2-induced LC3 up-regulation was mediated by heme oxygenase-1 and its by-product, carbon monoxide. Taken together, Nrf2 may be considered to play a role in cellular adaptive response to H. pylori-induced gastritis by inducing autophagy.Graphical abstractGraphical abstract for this article
  • Transport of ribavirin across the rat and human placental barrier: roles
           of nucleoside and ATP-binding cassette drug efflux transporters
    • Abstract: Publication date: Available online 2 February 2019Source: Biochemical PharmacologyAuthor(s): S. Karbanova, L. Cerveny, L. Jiraskova, R. Karahoda, M. Ceckova, Z. Ptackova, F. Staud Ribavirin is a broad-spectrum nucleoside-derived antiviral drug used in combination pharmacotherapy treatment of hepatitis C virus infection. Current evidence indicates that ribavirin-associated teratogenicity is not significant in humans, but more information about the developmental toxicity and mechanisms involved in ribavirin placental kinetics is required to assure its safe use in pregnancy. Thus, we have investigated potential roles of equilibrative nucleoside transporters (ENTs, SLC29A), Na+-dependent influx-mediating concentrative nucleoside transporters (CNTs, SLC28A), and ATP-binding cassette efflux pumps (ABC), in ribavirin placental pharmacokinetics. Our data indicate that ENT1 participates in uptake of ribavirin by BeWo cells, fresh human placental villous fragments and microvillous plasma membrane (MVM) vesicles while activity of CNTs (probably CNT2) was only observed in BeWo cells. In situ dual perfusion experiments with rat term placenta in an open circuit setup showed that ENT inhibition significantly decreases total ribavirin maternal-to-foetal and foetal-to-maternal clearances. In contrast, no contribution of ABC transporters, p-glycoprotein (ABCB1), breast cancer resistance protein (ABCG2), or multidrug resistance-associated protein (ABCC2) was detected in assays with MDCKII cells overexpressing them, or in closed circuit dual perfusion experiments with rat term placenta. In summary, our data show that ribavirin placental pharmacokinetics are largely controlled by ENT1 activity and independent of ABCB1, ABCG2, and ABCC2 efflux pumps.Graphical abstractGraphical abstract for this article
  • Dehydrocostus lactone (DHC) Suppresses Estrogen Deficiency-Induced
    • Abstract: Publication date: Available online 2 February 2019Source: Biochemical PharmacologyAuthor(s): Zhaoning Li, Guixin Yuan, Xixi Lin, Qian Liu, Jiake Xu, Zhen Lian, Fangming Song, Jinjian Zheng, Dantao Xie, Lingzi Chen, Xinjia Wang, Haotian Feng, Mengyu Zhou, Guanfeng Yao Osteoporosis is a chronic bone lytic disease, because of inadequate bone ossification and/or excessive bone resorption. Even though drugs are currently available for the treatment of osteoporosis, there remains an unmet need for the development of more specific novel agents with less adverse effects. Dehydrocostus lactone (DHC), a natural sesquiterpene lactone, was previously found to affect the differentiation of inflammatory cells by the inhibiting NF-κB pathways, and garnered much interest for its anti-cancer properties via SOCS-mediated cell cycle arrest and apoptosis. As NF-κB pathway plays an essential role in osteoclast differentiation, we sought to discover the biological effects of DHC on osteoclast differentiation and resorptive activity, as well as the underlying mechanisms on these effects. Our research found that DHC inhibited RANKL-induced osteoclast differentiation, bone resorption and osteoclast specific genes expression via suppression of NF-κB and NFAT signaling pathways in vitro. We further demonstrated that DHC protected against ovariectomy (OVX)-induced osteoprosis in the mouse model and the protective effects may be mediated through the attenuation of NF-κB signaling pathway. Thus, this study provides insight that DHC might be used as a potential pharmacological treatment for osteoporosis.Graphical abstractGraphical abstract for this article
  • Crocin, a carotenoid, suppresses spindle microtubule dynamics and
           activates the mitotic checkpoint by binding to tubulin
    • Abstract: Publication date: Available online 30 January 2019Source: Biochemical PharmacologyAuthor(s): Avishkar V. Sawant, Shalini Srivastava, Shweta S. Prassanawar, Bhabatarak Bhattacharyya, Dulal Panda Crocin, a constituent of the saffron spice, exhibits promising antitumor activity in animal models and also inhibits the proliferation of several types of cancer cells in culture. Recently, we have shown that crocin binds to purified tubulin at the vinblastine site, depolymerizes microtubules and induces a mitotic block in cultured cells. Here, we extend our previous suggestion and explore the cellular effects of crocin to further understand its mechanism of action. In a kinetic study, we observed that the crocin-induced depolymerization of microtubules preceded both DNA damage and reactive oxygen species generation indicating that depolymerizing microtubules is the primary action of crocin. Crocin also inhibited the growth of cold-depolymerized microtubules in HeLa cells indicating that it can inhibit microtubule dynamics. Using fluorescence recovery after photobleaching, crocin was found to suppress the spindle microtubule dynamics in live HeLa cells. Further, crocin treatment resulted in activation of spindle assembly checkpoint proteins, BubR1 and Mad2. Similar to other microtubule-targeting agents, crocin also perturbed the localization of end-binding protein EB1 from the growing microtubule ends and enhanced the acetylation of remaining microtubules. Further, crocin was found to bind to purified tubulin with a dissociation constant of 12 ± 1.5 μM. The results suggested that crocin exerted its antiproliferative effect primarily by inhibiting the assembly and dynamics of microtubules. Importantly, the combination of crocin with known anticancer agents like combretastatin A-4, cisplatin, doxorubicin or sorafenib, exerted a strong synergistic cytotoxic effect in HeLa cells indicating that crocin may enhance the effectiveness of other anticancer agents.Graphical abstractGraphical abstract for this article
  • Benproperine, an ARPC2 inhibitor, suppresses cancer cell migration and
           tumor metastasis
    • Abstract: Publication date: Available online 30 January 2019Source: Biochemical PharmacologyAuthor(s): Yae Jin Yoon, Young-Min Han, Jiyeon Choi, Yu-Jin Lee, Jieun Yun, Su-Kyung Lee, Chang Woo Lee, Jong Soon Kang, Seung-Wook Chi, Jeong Hee Moon, Sangku Lee, Dong Cho Han, Byoung-Mog Kwon Metastasis is the leading cause of cancer mortality and cancer cell migration is an essential stage of metastasis. We identified benproperine (Benp, a clinically used antitussive drug) as an inhibitor of cancer cell migration and an anti-metastatic agent. Benp selectively inhibited cancer cell migration and invasion, which also suppressed metastasis of cancer cells in animal models. Actin-related protein 2/3 complex subunit 2 (ARPC2) was identified as a molecular target of Benp by affinity column chromatography with Benp-tagged Sepharose beads. Benp bound directly to ARPC2 in cells, which was validated by pull-down assay using Benp-biotin and label-free biochemical methods such as the drug affinity responsive target stability (DARTS) and cellular thermal shift assay (CETSA). Benp inhibited Arp2/3 function, showing disruption of lamellipodial structure and inhibition of actin polymerization. Unlike Arp2/3 inhibitors, Benp selectively inhibited the migration of cancer cells but not normal cells. ARPC2-knockdown cancer cells showed defective cell migration and suppressed metastasis in an animal model. Therefore, ARPC2 is a potential target for anti-metastatic therapy, and Benp has the clinical potential to block metastasis. Furthermore, Benp is a useful agent for studying the functions of the Arp2/3 complex in cancer cell migration and metastasis.Graphical abstractGraphical abstract for this article
  • Ability of CP-532,903 to protect mouse hearts from ischemia/reperfusion
           injury is dependent on expression of A3 adenosine receptors in
    • Abstract: Publication date: Available online 30 January 2019Source: Biochemical PharmacologyAuthor(s): Tina C. Wan, Akihito Tampo, Wai-Meng Kwok, John A. Auchampach A3 adenosine receptor (A3AR) agonists are effective at limiting injury caused by ischemia/reperfusion injury of the heart in experimental animal models. However, understanding of their mechanism of action, which is likely multifactorial, remains incomplete. In prior studies, it has been demonstrated that A3AR-mediated ischemic protection is blocked by glibenclamide and is absent in Kir6.2 gene ablated mice that lack the pore-forming subunit of the ATP-sensitive potassium (KATP) channel, suggesting one contributing mechanism may involve accelerated activation of KATP channels. However, presence of A3ARs in the myocardium has yet to be established. Utilizing a whole-cell recording technique, in this study we confirm functional expression of the A3AR in adult mouse ventricular cardiomyocytes, coupled to activation of ATP-dependent potassium (KATP) channels via Gi inhibitory proteins. We further show that ischemic protection provided by the selective A3AR agonist CP-532,903 in an isolated, buffer-perfused heart model is lost completely in Adora3LoxP/LoxP;Myh6-Cre mice, which is a newly developed model developed and comprehensively described herein whereby the A3AR gene (Adora3) is deleted exclusively in cardiomyocytes. Our findings, taken together with previously published work, are consistent with the hypothesis that A3AR agonists provide ischemic tolerance, at least in part, by facilitating opening of myocardial KATP channels.Graphical abstractGraphical abstract for this article
  • Paricalcitol alleviates lipopolysaccharide-induced depressive-like
           behavior by suppressing hypothalamic microglia activation and
    • Abstract: Publication date: Available online 28 January 2019Source: Biochemical PharmacologyAuthor(s): Ming-chao He, Zhe Shi, Nan-nan Sha, Nan Chen, Shi-yu Peng, Duan-fang Liao, Man-sau Wong, Xiao-li Dong, Yong-jun Wang, Ti-fei Yuan, Yan Zhang Depression is highly prevalent in patients suffering from chronic inflammatory diseases. Dysregulated neuroinflammation and concomitant activated micorglia play a pivotal role in the pathogenesis of depression. Paricalcitol (Pari), a vitamin D2 analogue, has been demonstrated to exert anti-inflammative effects on renal and cardiovascular diseases. In this study, mice were pretreated with Pari before being induced to acute depression-like behaviors by systemic lipopolysaccharide (LPS) injection. To determine the therapeutic effects of Pari, alterations in acute body weight, sucrose preference, forced swimming and tail suspension tests were assessed. Then, alterations of pro-inflammation cytokine IL1-β level and microglia activity in the hypothalamus, which are involved in the pathophysiology of depression, were examined. The results showed that Pari significantly alleviated systemic LPS injection induced depressive-like behaviors as shown by increased sucrose preference and decreased TST and FST immobility. Pari could specifically regulate microglia-mediated neuroinflammation process and local activity of renin-angiotensin system to exert its anti-depressant effects. This study demonstrated a potential for paricalcitol in treating depressive symptoms induced by systemic inflammation, particularly in patients with chronic hypertension.Graphical abstractGraphical abstract for this article
  • Mining the NaV1.7 interactome: Opportunities for chronic pain
    • Abstract: Publication date: Available online 27 January 2019Source: Biochemical PharmacologyAuthor(s): Lindsey A. Chew, Shreya S. Bellampalli, Erik T. Dustrude, Rajesh Khanna The peripherally expressed voltage-gated sodium NaV1.7 (gene SCN9A) channel boosts small stimuli to initiate firing of pain-signaling dorsal root ganglia (DRG) neurons and facilitates neurotransmitter release at the first synapse within the spinal cord. Mutations in SCN9A produce distinct human pain syndromes. Widely acknowledged as a “gatekeeper” of pain, NaV1.7 has been the focus of intense investigation but, to date, no NaV1.7-selective drugs have reached the clinic. Elegant crystallographic studies have demonstrated the potential of designing highly potent and selective NaV1.7 compounds but their therapeutic value remains untested. Transcriptional silencing of NaV1.7 by a naturally expressed antisense transcript has been reported in rodents and humans but whether this represents a viable opportunity for designing NaV1.7 therapeutics is currently unknown. The demonstration that loss of NaV1.7 function is associated with upregulation of endogenous opioids and potentiation of mu- and delta-opioid receptor activities, suggests that targeting only NaV1.7 may be insufficient for analgesia. However, the link between opioid-dependent analgesic mechanisms and function of sodium channels and intracellular sodium-dependent signaling remains controversial and disputed. Thus, additional new targets - regulators, modulators - are needed. In this context, we mine the literature for the known interactome of NaV1.7 with a focus on protein interactors that affect the channel’s trafficking or link it to opioid signaling. As a case study, we present antinociceptive evidence of allosteric regulation of NaV1.7 by the cytosolic collapsin response mediator protein 2 (CRMP2). Throughout discussions of these possible new targets, we offer thoughts on the therapeutic implications of modulating NaV1.7 function in chronic pain.Graphical abstractGraphical abstract for this article
  • Understanding the Cancer Stem Cell Phenotype: A Step Forward in the
           Therapeutic Management of Cancer
    • Abstract: Publication date: Available online 25 January 2019Source: Biochemical PharmacologyAuthor(s): Andrea Li Ann Wong, Gregory Lucien Bellot, Jayshree L. Hirpara, Shazib Pervaiz The experimental validation of the existence of cancer stem cells (CSC) has had a significant impact on our understanding of the cellular mechanisms and signaling networks involved in the process of carcinogenesis and its progression. These findings provide insights into the critical role that tumor microenvironment and metabolism play in the acquisition of the drug resistance phenotype as well as provide potential targets for therapeutic exploitation. Here we briefly review the literature on the involvement of key signaling pathways such as Wnt/β-catenin, Notch, Hedgehog and STAT3 in the appearance of cancer cells with stem cells-like characteristics. In addition, we also highlight some of the recent therapeutic strategies used to target these pathways as well as approaches aiming to specifically target CSCs through their distinctive metabolic features.Graphical abstractGraphical abstract for this article
  • A Delicate Balance - The BCL-2 Family and its Role in Apoptosis,
           Oncogenesis, and Cancer Therapeutics
    • Abstract: Publication date: Available online 19 January 2019Source: Biochemical PharmacologyAuthor(s): Tristan Knight, Daniel Luedtke, Holly Edwards, Jeffrey W. Taub, Yubin Ge Evasion of apoptosis is fundamental to the pathogenesis of cancer. Members of the B-cell Lymphoma 2 (BCL-2) protein family are key pro- and anti-apoptotic regulators, and in healthy cells are held in a fine, delicate balance – perturbations of which may tip a cell irreversibly towards cellular death or, conversely, allow a cell to permanently escape apoptosis and immortalize itself as a malignant clone. The restoration of this balance or, indeed, adjustment in favor of apoptosis via manipulation of the BCL-2 family, is a promising area in the realm of molecular therapeutics, and one in which breathtaking advances are currently being made. The purpose of this review is to outline the role of the BCL-2 family in apoptosis, to contrast its optimal functioning with those disruptions seen in malignancy, and to provide an overview of the medications both presently available and currently under development which selectively target members of this family.Graphical abstractGraphical abstract for this article
  • Targeting intrinsic cell death pathways to control fungal pathogens
    • Abstract: Publication date: Available online 17 January 2019Source: Biochemical PharmacologyAuthor(s): Madhura Kulkarni, Zachary D. Stolp, J. Marie Hardwick Fungal pathogens pose an increasing threat to public health. Limited clinical drug regimens and emerging drug-resistant isolates challenge infection control. The global burden of human fungal pathogens is estimated at 1 billion infections and 1.5 million deaths annually. In addition, plant fungal pathogens increasingly threaten global food resources. Novel strategies are needed to combat emerging fungal diseases and pan-resistant fungi. An untapped mechanistically novel approach is to pharmacologically activate the intrinsic cell death pathways encoded by pathogenic fungi. This strategy is analogous to new anti-cancer therapeutics now entering the clinic. Here we summarize the best understood examples of cell death mechanisms encoded by pathogenic fungi, contrast these to mammalian cell death pathways, and highlight the gaps in knowledge towards identifying potential death effectors as druggable targets.Graphical abstractFungal species appear to undergo forms of programmed cell death though the detailed mechanisms are not yet known.Graphical abstract for this article
  • Non-immunological toxicological mechanisms of metamizole-associated
           neutropenia in HL60 cells
    • Abstract: Publication date: Available online 14 January 2019Source: Biochemical PharmacologyAuthor(s): Deborah Rudin, Angelo Lanzilotto, Fabio Bachmann, Catherine E Housecroft, Edwin C Constable, Jürgen Drewe, Manuel Haschke, Stephan Krähenbühl Metamizole is an analgesic and antipyretic, but can cause neutropenia and agranulocytosis. We investigated the toxicity of the metabolites N-methyl-4-aminoantipyrine (MAA), 4-aminoantipyrine (AA), N-formyl-4-aminoantipyrine (FAA) and N-acetyl-4-aminoantipyrine (AAA) on neutrophil granulocytes and on HL60 cells (granulocyte precursor cell line). MAA, FAA, AA, and AAA (up to 100 µM) alone were not toxic for HL60 cells or granulocytes. In the presence of the myeloperoxidase substrate H2O2, MAA reduced cytotoxicity for HL60 cells at low (
  • Anti-tumor effect of sulfasalazine in neuroblastoma
    • Abstract: Publication date: Available online 9 January 2019Source: Biochemical PharmacologyAuthor(s): Marie R. Mooney, Dirk Geerts, Eric J. Kort, André S. Bachmann Neuroblastoma (NB) is a tumor arising from the sympathetic nervous system during infancy and early childhood. High-risk patients who relapse often fail to respond to further therapy, which results in 5-year survival rate for this patient group below 5%. Therefore, there continues to be an urgent need for innovative treatments. Recently, we found that sulfasalazine (SSZ), an FDA-approved drug for the treatment of rheumatoid arthritis and ulcerative colitis induces anti-proliferative effects in NB tumor cells. SSZ was recently shown to inhibit sepiapterin reductase (SPR), a key enzyme that produces tetrahydrobiopterin (BH4) in the nitric oxide (NO) pathway. Here we tested SSZ against purified SPR in vitro, measured the anti-proliferative effect of SSZ on a panel of MYCN amplified and MYCN non-amplified NB cell lines, and assessed the anti-tumor effect of SSZ in NB tumor-xenografted mice. We found that the expression of both SPR mRNA and SPR protein was significantly higher in cell lines without MYCN amplification. SSZ inhibited SPR enzyme activity in vitro and exhibits anti-proliferative activity in a large number of NB cell lines derived from high-risk tumors. Importantly, oral/intraperitoneal (i.p.) SSZ co-administration resulted in measureable anti-tumor effects in vivo. The FDA-approved drug SSZ, a well-tolerated drug in clinical use, could be repositioned to inhibit tumor growth in NB.Graphical abstractGraphical abstract for this article
  • Potentialization of anticancer agents by identification of new
           chemosensitizers active under hypoxia
    • Abstract: Publication date: Available online 7 January 2019Source: Biochemical PharmacologyAuthor(s): MARX Sébastien, VAN GYSEL Mégane, BREUER Aurélie, DAL MASO Thomas, MICHIELS Carine, WOUTERS Johan, LE CALVE Benjamin Hypoxia is one of the most important biological phenomena that influences cancer agressiveness and chemotherapy resistance. Cancer cells display dysregulated pathways notably resulting from oncogene expression. Tumors also show modifications in extracellular pH, extracellular matrix remodeling, neo-angiogenesis, hypoxia compared to normal tissues. Classically, the conventional anticancer therapies are efficient in cancer cells in normoxic conditions but under hypoxia, chemoresistance may occur. The addition of compounds that potentiate their activity in low oxygen environment could be a strategy to counteract this resistance. To identify new compounds active in hypoxia, we screened one hundred molecules with different chemical structures from an internal chemolibrary. Their potential ability to increase the activity of taxol and etoposide independently of their mechanism of action has been assayed. After a first step of selection, based on biological/pharmacological properties and chemical structure analysis, we identified three potential hits. Two hits are closely related amides/ureas and the third is a thiosemicarbazone. The compounds present no activity in cancer and normal cells when used alone but demonstrate chemosensitizing activity under hypoxia. Finally, by analyzing cell death, the indole thiosemicarbazone was shown to be able to significantly potentiate apoptosis induced by taxol and etoposide in two models of cancer cell lines. This new compound could lead to the development of an original series of chemosensitizers active under hypoxia.Graphical abstractGraphical abstract for this article
  • Natural modulators of the hallmarks of immunogenic cell death
    • Abstract: Publication date: Available online 5 January 2019Source: Biochemical PharmacologyAuthor(s): Flavia Radogna, Mario Dicato, Marc Diederich Natural compounds act as immunoadjuvants as their therapeutic effects trigger cancer stress response and release of damage-associated molecular patterns (DAMPs). These reactions occur through an increase in the immunogenicity of cancer cells that undergo stress followed by immunogenic cell death (ICD). These processes result in a chemotherapeutic response with a potent immune-mediating reaction. Natural compounds that induce ICD may function as an interesting approach in converting cancer into its own vaccine. However, multiple parameters determine whether a compound can act as an ICD inducer, including the nature of the inducer, the premortem stress pathways, the cell death pathways, the intrinsic antigenicity of the cell, and the potency and availability of an immune cell response. Thus, the identification of hallmarks of ICD is important in determining the prognostic biomarkers for new therapeutic approaches and combination treatments.Graphical abstractGraphical abstract for this article
  • Tumor cell escape from therapy-induced senescence
    • Abstract: Publication date: Available online 19 December 2018Source: Biochemical PharmacologyAuthor(s): Tareq Saleh, Liliya Tyutyunyk-Massey, Graeme F. Murray, Moureq R. Alotaibi, Ajinkya S. Kawale, Zeinab Elsayed, Scott C. Henderson, Vasily Yakovlev, Lynne W. Elmore, Amir Toor, Hisashi Harada, Jason Reed, Joseph W. Landry, David A. Gewirtz H460 non-small cell lung, HCT116 colon and 4T1 breast tumor cell lines induced into senescence by exposure to either etoposide or doxorubicin were able to recover proliferative capacity both in mass culture and when enriched for the senescence-like phenotype by flow cytometry (based on β-galactosidase staining and cell size, and a senescence-associated reporter, BTG1-RFP). Recovery was further established using both real-time microscopy and High-Speed Live-Cell Interferometry (HSLCI) and was shown to be accompanied by the attenuation of the senescence-associated secretory phenotype (SASP). Cells enriched for the senescence-like phenotype were also capable of forming tumors when implanted in both immunodeficient and immunocompetent mice. As chemotherapy-induced senescence has been identified in patient tumors, our results suggest that certain senescence-like phenotypes may not reflect a terminal state of growth arrest, as cells that recover with self-renewal capacity may ultimately contribute to disease recurrence.Graphical abstractGraphical abstract for this article
  • Contrasting effects of microtubule destabilizers versus stabilizers on
           induction of death in G1 phase of the cell cycle
    • Abstract: Publication date: Available online 19 December 2018Source: Biochemical PharmacologyAuthor(s): Magdalena Delgado, Alicja Urbaniak, Timothy C. Chambers Microtubule targeting agents (MTAs) have been reported to manifest their cytotoxic effects not only in mitosis but also in interphase. However, the relationship between phase-specific susceptibility and MTA concentration, especially with respect to microtubule integrity, remains poorly defined. In addition, whether microtubule stabilizers and destabilizers act similarly or differ in the ability to induce interphase death is unclear. In order to resolve these uncertainties, we report here the results of a systematic comparison of primary acute lymphoblastic leukemia (ALL) and HeLa cells treated with three different MTAs, namely the microtubule stabilizer paclitaxel and two microtubule destabilizers, vincristine, and eribulin. Both types of cells were sensitive to each MTA, with IC50 values in the sub-nanomolar to low nanomolar range. Primary ALL cells arrested in mitosis when treated with paclitaxel at all tested concentrations, whereas the effects of vincristine or eribulin were concentration-dependent; low (< 30 nM) concentrations induced mitotic death whereas higher concentrations (>100 nM) induced death directly in G1 phase. G1 phase death in response to higher concentrations of the destabilizers was associated with complete loss of interphase microtubule structure. In contrast, HeLa cells were only susceptible in M phase regardless of drug type or concentration. These results represent an important advance in our understanding and appreciation of microtubule function, and indicate that susceptibility to MTAs in G1 phase is both cell type- and drug type-restricted. The findings have important implications for the clinical use of MTAs especially in the context of drug combinations.Graphical abstractGraphical abstract for this article
  • Loperamide overcomes the resistance of colon cancer cells to bortezomib by
           inducing CHOP-mediated paraptosis-like cell death
    • Abstract: Publication date: Available online 7 December 2018Source: Biochemical PharmacologyAuthor(s): In Young Kim, Min June Shim, Dong Min Lee, A Reum Lee, Mi Ae Kim, Mi Jin Yoon, Mi Ri Kwon, Hae In Lee, Min Ji Seo, Yong Won Choi, Kyeong Sook Choi Although the proteasome inhibitor (PI) bortezomib (Btz) is in current clinical use as a front-line treatment for multiple myeloma, its clinical efficacy in solid tumors has not been satisfactory. Here, we show that loperamide (Lop), an antidiarrheal drug, effectively sensitizes various colon cancer cells, but not normal epithelial cells, to PI-mediated cell death. We report that combined treatment with Btz and Lop induces paraptosis-like cell death accompanied by severe endoplasmic reticulum (ER)-derived vacuolation. Furthermore, Lop potentiates Btz-mediated ER stress and ER dilation due to misfolded protein accumulation and Ca2+ imbalance, leading to CHOP upregulation and subsequent paraptosis-like cell death. Taken together, our results show for the first time that a combined regimen of PI and Lop may provide an effective and safe therapeutic strategy against solid tumors, including colon cancer, by enhancing the sensitivity to PIs and reducing the side effects of such treatment.Graphical abstractGraphical abstract for this article
  • BubR1 depletion delays apoptosis in the microtubule-depolymerized cells
    • Abstract: Publication date: Available online 22 November 2018Source: Biochemical PharmacologyAuthor(s): Afsana Naaz, Shazia Ahad, Ankit Rai, Avadhesha Surolia, Dulal Panda We investigated the role of a spindle assembly checkpoint protein, BubR1, in determining the mechanism of cell killing of an anti-microtubule agent CXI-benzo-84. CXI-benzo-84 dampened microtubule dynamics in live MCF-7 cells. The compound arrested MCF-7 cells in mitosis and induced apoptosis in these cells. Though CXI-benzo-84 efficiently depolymerized microtubules in the BubR1-depleted MCF-7 cells, it did not arrest the BubR1-depleted cells at mitosis. Interestingly, apoptosis occurred in the BubR1-depleted MCF-7 cells in the absence of a mitotic block suggesting that the mitotic block is not a prerequisite for the induction of apoptosis by anti-microtubule agents. In the presence of CXI-Benzo-84, the level of apoptosis was initially found to be lesser in the BubR1-depleted MCF-7 cells than the control cells; however, the BubR1-depleted cells displayed a similar level of apoptosis as the control cells at 72 hours of drug treatment. The depletion of BubR1 enhanced DNA damage in MCF-7 cells upon microtubule depolymerization. In addition, CXI-benzo-84 in combination with cisplatin-induced more cell death in BubR1-depleted cells than the BubR1-expressing MCF-7 cells. The results indicated a possibility that the BubR1-compromised cancer patients can be treated with combination therapy.Graphical abstractGraphical abstract for this article
  • Clomiphene citrate induces nuclear translocation of the TFEB transcription
           factor and triggers apoptosis by enhancing lysosomal membrane
    • Abstract: Publication date: Available online 22 November 2018Source: Biochemical PharmacologyAuthor(s): Wei Li, Jieru Lin, Zhichen Shi, Jiren Wen, Yuyin Li, Zhenxing Liu, Aipo Diao The autophagy-lysosome pathway plays a central role in cellular homeostasis by regulating the cellular degradative machinery. The transcription factor EB (TFEB) regulates the biogenesis and function of both lysosomes and autophagosomes, and enhancement of TFEB function has emerged as an attractive therapeutic strategy for lysosome-related disorders. However, little is known about the role of TFEB activation in regulating the cellular fate. Here, we describe that clomiphene citrate (CC), a selective estrogen receptor modulator, promotes nuclear translocation of TFEB and increases lysosomal biogenesis in HeLa and MDA-MB-231 cells. Treatment with CC inhibits cell viability and causes apoptosis by increasing the release of proteases cathepsin B (CatB) and cathepsin D (CatD) from lysosomes into the cytosol. In contrast, knockdown of TFEB rescues the cells from CC-induced cell death. Furthermore, CC-induced TFEB activation also enhances the autophagy flux in HeLa cells. Knockdown of autophagy-related gene 7 (ATG7) significantly decreases the CC-induced CatB and CatD release and cell death, suggesting that autophagy contributes to the lysosomal membrane permeabilization (LMP) caused by CC. Altogether, these findings have broad implications for our understanding of TFEB function and provide new insights into CC pharmacological therapy.Graphical abstractGraphical abstract for this article
  • The roles of ubiquitination in extrinsic cell death pathways and its
           implications for therapeutics
    • Abstract: Publication date: Available online 16 November 2018Source: Biochemical PharmacologyAuthor(s): Jinho Seo, Min Wook Kim, Kwang-Hee Bae, Sang Chul Lee, Jaewhan Song, Eun-Woo Lee Regulation of cell survival and death, including apoptosis and necroptosis, is important for normal development and tissue homeostasis, and disruption of these processes can cause cancer, inflammatory diseases, and degenerative diseases. Ubiquitination is a cellular process that induces proteasomal degradation by covalently attaching ubiquitin to the substrate protein. In addition to proteolytic ubiquitination, nonproteolytic ubiquitination, such as M1-linked and K63-linked ubiquitination, has been shown to be important in recent studies, which have demonstrated its function in cell signaling pathways that regulate inflammation and cell death pathways. In this review, we summarize the TRAIL- and TNF-induced death receptor signaling pathways along with recent advances in this field and illustrate how different types of ubiquitination control cell death and survival. In particular, we provide an overview of the different types of ubiquitination, target residues, and modifying enzymes, including E3 ligases and deubiquitinating enzymes. Given the relevance of these regulatory pathways in human disease, we hope that a better understanding of the regulatory mechanisms of cell death pathways will provide insights into and therapeutic strategies for related diseases.Graphical abstractGraphical abstract for this article
  • Emerging roles of ADP-ribosyl-acceptor hydrolases (ARHs) in tumorigenesis
           and cell death pathways
    • Abstract: Publication date: Available online 27 September 2018Source: Biochemical PharmacologyAuthor(s): Xiangning Bu, Jiro Kato, Joel Moss Malignant transformation may occur in the background of post-translational modification, such as ADP-ribosylation, phosphorylation and acetylation. Recent genomic analysis of ADP-ribosylation led to the discovery of more than twenty ADP-ribosyltransferases (ARTs), which catalyze either mono- or poly-ADP-ribosylation. ARTs catalyze the attachment of ADP-ribose to acceptor molecules. The ADP-ribose-acceptor bond can then be cleaved by a family of hydrolases in a substrate-specific manner, which is dependent on the acceptor and its functional group, e.g., arginine (guanidino), serine (hydroxyl), aspartate (carboxyl). These hydrolases vary in structure and function, and include poly-ADP-ribose glycohydrolase (PARG), MacroD1, MacroD2, terminal ADP-ribose protein glycohydrolase 1 (TARG1) and ADP-ribosyl-acceptor hydrolases (ARHs). In murine models, PARG deficiency increased susceptibility to alkylating agents-induced carcinogenesis. Similarly, by cleaving mono-ADP-ribosylated arginine on target proteins, ARH1 appears to inhibit tumor formation, suggesting that ARH1 is a tumor-suppressor gene. Although ARH3 is similar to ARH1 in amino acid sequence and crystal structure, ARH3 does not cleave ADP-ribose-arginine, rather it degrades in an exocidic manner, the PAR polymer and cleaves O-acetyl-ADP-ribose (OAADPr) and the ADP-ribose-serine linkage in acceptor proteins. Under conditions of oxidative stress, ARH3-deficient cells showed increased cytosolic PAR accumulation and PARP-1 mediated cell death. These findings expand our understanding of ADP-ribosylation and provide new therapeutic targets for cancer treatment. In the present review, research on ARH1-regulated tumorigenesis and cell death pathways that are enhanced by ARH3 deficiency are discussed.Graphical abstractGraphical abstract for this article
  • About canonical, non-canonical and immunogenic cell death: Basic
           mechanisms and translational applications: A meeting report of the
           International Cell Death Society
    • Abstract: Publication date: Available online 14 September 2018Source: Biochemical PharmacologyAuthor(s): Zahra Zakeri, Richard A. Lockshin, Marc Diederich International Cell Death Society held its 25th meeting, entitled “About canonical, non-canonical, and immunogenic cell death: basic mechanisms and translational applications” in Seoul, South Korea, May 31-June 2, 2018, addressed the most current issues in the field. Now that many types and pathways of cell death are recognized, attention has turned to how the threshold to death is maintained or surpassed, and how and what intracellular signals control the process. Most of the speakers addressed these topics, focusing on mitochondria and on new high-resolution techniques that promise to answer current questions.
  • Tofacitinib and TPCA-1 exert chondroprotective effects on extracellular
           matrix turnover in bovine articular cartilage ex vivo
    • Abstract: Publication date: Available online 29 July 2018Source: Biochemical PharmacologyAuthor(s): Cecilie F. Kjelgaard-Petersen, Neha Sharma, Ashref Kayed, Morten A. Karsdal, Ali Mobasheri, Per Hägglund, Anne-Christine Bay-Jensen, Christian S. Thudium ObjectiveCurrently, there are no disease-modifying osteoarthritis drugs (DMOADs) approved for osteoarthritis. It is hypothesized that a subtype of OA may be driven by inflammation and may benefit from treatment with anti-inflammatory small molecule inhibitors adopted from treatments of rheumatoid arthritis. This study aimed to investigate how small molecule inhibitors of intracellular signaling modulate cartilage degradation and formation as a pre-clinical model for structural effects.DesignBovine cartilage explants were cultured with oncostatin M (OSM) and tumour necrosis factor α (TNF-α) either alone or combined with the small molecule inhibitors: SB203580 (p38 inhibitor), R406 (Spleen tyrosine kinase (Syk) inhibitor), TPCA-1 (Inhibitor of κB kinase (Ikk) inhibitor), or Tofacitinib (Tofa) (Janus kinases (Jak) inhibitor). Cartilage turnover was assessed with the biomarkers of degradation (AGNx1 and C2M), and type II collagen formation (PRO-C2) using ELISA. Explant proteoglycan content was assessed by Safranin O/Fast Green staining.ResultsR406, TPCA-1 and Tofa reduced the cytokine-induced proteoglycan loss and decreased AGNx1 release 3.7-, 43- and 32-fold, respectively. SB203580 showed no effect. All inhibitors suppressed C2M at a concentration of 3 µM. TPCA-1 and Tofa increased the cytokine reduced PRO-C2 3.5 and 3.7-fold, respectively.ConclusionUsing a pre-clinical model we found that the inhibitors TPCA-1 and Tofa inhibited cartilage degradation and rescue formation of type II collagen under inflammatory conditions, while R406 and SB203580 only inhibited cartilage degradation, and SB203580 only partially. These pre-clinical data suggest that TPCA-1 and Tofa preserve and help maintain cartilage ECM under inflammatory conditions and could be investigated further as DMOADs for inflammation-driven osteoarthritis.Graphical abstractGraphical abstract for this article
  • Dedicated to the memory of Professor Jacques Gielen
    • Abstract: Publication date: Available online 5 December 2005Source: Biochemical PharmacologyAuthor(s): Jacques Piette
  • Note from the Publisher
    • Abstract: Publication date: Available online 5 December 2005Source: Biochemical PharmacologyAuthor(s):
  • Poly(ADP-ribosylated) proteins in β-amyloid peptide-stimulated
           microglial cells
    • Abstract: Publication date: Available online 9 November 2018Source: Biochemical PharmacologyAuthor(s): Virginia Correani, Sara Martire, Giuseppina Mignogna, Lisa Beatrice Caruso, Italo Tempera, Alessandra Giorgi, Maddalena Grieco, Luciana Mosca, M.Eugenia Schininà, Bruno Maras, Maria d'Erme Amyloid-treated microglia prime and sustain neuroinflammatory processes in the central nervous system activating different signalling pathways inside the cells. Since a key role for PARP-1 has been demonstrated in inflammation and in neurodegeneration, we investigated PARylated proteins in resting and in β-amyloid peptide treated BV2 microglial cells. A total of 1158 proteins were identified by mass spectrometry with 117 specifically modified in the amyloid-treated cells. Intervention of PARylation on the proteome of microglia showed to be widespread in different cellular districts and to affect various cellular pathways, highlighting the role of this dynamic post-translational modification in cellular regulation. Ubiquitination is one of the more enriched pathways, encompassing PARylated proteins like NEDD4, an E3 ubiquitine ligase and USP10, a de-ubiquitinase, both associated with intracellular responses induced by β-amyloid peptide challenge. PARylation of NEDD4 may be involved in the recruiting of this protein to the plasma membrane where it regulates the endocytosis of AMPA receptors, whereas USP10 may be responsible for the increase of p53 levels in amyloid stimulated microglia. Unfolded Protein Response and Endoplasmic Reticulum Stress pathways, strictly correlated with the Ubiquitination process, also showed enrichment in PARylated proteins. PARylation may thus represent one of the molecular switches responsible for the transition of microglia towards the inflammatory microglia phenotype, a pivotal player in brain diseases including neurodegenerative processes. The establishment of trials with PARP inhibitors to test their efficacy in the containment of neurodegenerative diseases may be envisaged.Graphical abstractGraphical abstract for this article
  • Dormant, quiescent, tolerant and persister cells: four synonyms for the
           same target in cancer
    • Abstract: Publication date: Available online 9 November 2018Source: Biochemical PharmacologyAuthor(s): François M. Vallette, Christophe Olivier, Frédéric Lézot, Lisa Oliver, Denis Cochonneau, Lisenn Lalier, Pierre-François Cartron, Dominique Heymann Although many drugs/treatments are now available for most diseases, too often, resistance to these treatments impedes complete therapeutic success. Acquired resistance is a major problem in many pathologies but it is an acute one in cancers and infections. This is probably because these diseases often require long durations of treatment, which ascribe to the selection of resistant cells. However, the actual mechanisms implicated in the selection process are still under debate. It is becoming increasingly clear that resistance is associated with the heterogeneity of cancer cells or micro-organisms and that multiple mechanisms underlie the emergence of drug-resistant subpopulations. Recently, it has been suggested that a subpopulation of drug tolerant cells present in cancer populations and called “persisters” play a major role in this resistance. Recent studies have shown that microorganisms share similar properties. Still, how persister/tolerant cells intervene in the development of resistance is not completely elucidated but seems to be related to epigenetic changes in treated cells and the capacity of persisters to modulate and/or highjack their microenvironment. Due to the complexity of this process, the input from mathematicians, as well as new methods of bioinformatics and statistics, is necessary to fully comprehend the acquisition of resistance/tolerance deriving from and leading to the heterogeneous cell populations. The present review will give a brief overview of the most recent data available on drug tolerant cells in cancers and their similarities with microorganisms.Graphical abstractGraphical abstract for this article
  • Non-mitotic effect of albendazole triggers apoptosis of human leukemia
           cells via SIRT3/ROS/p38 MAPK/TTP axis-mediated TNF-α upregulation
    • Abstract: Publication date: Available online 7 November 2018Source: Biochemical PharmacologyAuthor(s): Liang-Jun Wang, Yuan-Chin Lee, Chia-Hui Huang, Yi-Jun Shi, Ying-Jung Chen, Sung-Nan Pei, Yu-Wei Chou, Long-Sen Chang Albendazole (ABZ) is a microtubule-targeting anthelmintic that acts against a variety of human cancer cells, but the dependence of its cytotoxicity on non-mitotic effect remains elusive. Thus, we aimed to explore the mechanistic pathway underlying the cytotoxicity of ABZ in human leukemia U937 cells. ABZ-induced apoptosis of U937 cells was characterized by mitochondrial ROS generation, p38 MPAK activation, TNF-α upregulation and activation of the death receptor-mediated pathway. Meanwhile, ABZ induced tubulin depolymerization and G2/M cell cycle arrest. ABZ-induced SIRT3 degradation elicited ROS-mediated p38 MAPK activation, leading to pyruvate kinase M2-mediated tristetraprolin (TTP) degradation. Inhibition of TTP-mediated TNF-α mRNA decay elicited TNF-α upregulation in ABZ-treated cells. Either the overexpression of SIRT3 or abolishment of ROS/p38 MAPK activation suppressed TNF-α upregulation and rescued the viability of ABZ-treated cells. In contrast to the inhibition of ROS/p38 MAPK pathway, SIRT3 overexpression attenuated tubulin depolymerization and G2/M arrest in ABZ-treated cells. Treatment with a SIRT3 inhibitor induced TNF-α upregulation and cell death without the induction of G2/M arrest in U937 cells. Taken together, our data indicate that ABZ-induced SIRT3 downregulation promotes its microtubule-destabilizing effect, and that the non-mitotic effect of ABZ largely triggers apoptosis of U937 cells via SIRT3/ROS/p38 MAPK/TTP axis-mediated TNF-α upregulation. Notably, the same pathway is involved in the ABZ-induced death of HL-60 cells.Graphical abstractGraphical abstract for this article
  • MDM2 and Mitochondrial Function: One Complex Intersection
    • Abstract: Publication date: Available online 1 November 2018Source: Biochemical PharmacologyAuthor(s): Camila Rubio-Patiño, Andrew Paul Trotta, Jerry Edward Chipuk Decades of research reveal that MDM2 participates in cellular processes ranging from macro-molecular metabolism to cancer signaling mechanisms. Two recent studies uncovered a new role for MDM2 in mitochondrial bioenergetics. Through the negative regulation of NDUFS1 (NADH:ubiquinone oxidoreductase 75 kDa Fe-S protein 1) and MT-ND6 (NADH dehydrogenase 6), MDM2 decreases the function and efficiency of Complex I (CI). These observations propose several important questions: (1) Where does MDM2 affect CI activity? (2) What are the cellular consequences of MDM2-mediated regulation of CI? (3) What are the physiological implications of these interactions? Here, we will address these questions and position these observations within the MDM2 literature.Graphical abstractGraphical abstract for this article
  • Hedgehog and Wingless signaling are not essential for autophagy-dependent
           cell death
    • Abstract: Publication date: Available online 26 October 2018Source: Biochemical PharmacologyAuthor(s): Tianqi Xu, Donna Denton, Sharad Kumar Autophagy-dependent cell death is a distinct mode of regulated cell death required in a context specific manner. One of the best validated genetic models of autophagy-dependent cell death is the removal of the Drosophila larval midgut during larval-pupal transition. We have previously shown that down-regulation of growth signaling is essential for autophagy induction and larval midgut degradation. Sustained growth signaling through Ras and PI3K blocks autophagy and consequently inhibits midgut degradation. In addition, the morphogen Dpp plays an important role in regulating the correct timing of midgut degradation. Here we explore the potential roles of Hh and Wg signaling in autophagy dependent midgut cell death. We demonstrate that Hh and Wg signaling are not involved in the regulation of autophagy-dependent cell death. However, surprisingly we found that one key component of these pathways, the Drosophila Glycogen Synthase Kinase 3, Shaggy (Sgg), may regulate midgut cell size independent of Hh and Wg signaling.Graphical abstractGraphical abstract for this article
  • Vincristine Ablation of Sirt2 Induces Cell Apoptosis and Mitophagy via
           Hsp70 Acetylation in MDA-MB-231 Cells
    • Abstract: Publication date: Available online 21 October 2018Source: Biochemical PharmacologyAuthor(s): Fanghui Sun, Xiaoxiao Jiang, Xuan Wang, Yong Bao, Guize Feng, Huijuan Liu, Xinhui Kou, Qing Zhu, Lan Jiang, Yonghua Yang Cancer cells are continuously challenged by adverse environmental stress and adopt diverse strategies to survive. Hsp70 plays pivotal roles in invasion, migration, drug resistance, and the survival of tumor cells. Hsp70 functions as molecular chaperone to protect tumor cells from stress-induced cell death. Hsp70 acetylation alters its chaperone activity in cell death pathways, but its relevance in the process of cell death and the underlying mechanisms involved are not well understood. In this study, we demonstrated that vincristine induces mitophagy via the disruption of Hsp70 binding with Sirt2, leading to Hsp70 acetylation at K126 and elevated sequestration of Bcl2 by Hsp70 for autophagosome creation. Acetylation at K126 significantly changes the physiological function of Hsp70 compared to acetylation at other sites. It also attenuates the protein folding and renaturation function of Hsp70 by altering the binding co-chaperones. In addition, acetylation at K126 inhibits Hsp70-mediated tumor cell invasion and migration, and the binding of Hsp70 to AIF1 and Apaf1 for promoting mitochondrial-mediated apoptosis. In conclusion, this study describes the molecular mechanism of vincristine induction of cell apoptosis and mitophagy via ablation of Sirt2 induced Hsp70 acetylation at K126 in MDA-MB-231 cells.Graphical abstractGraphical abstract for this article
  • Puerarin reverses cadmium-induced lysosomal dysfunction in primary rat
           proximal tubular cells via inhibiting Nrf2 pathway
    • Abstract: Publication date: Available online 19 October 2018Source: Biochemical PharmacologyAuthor(s): Li-Yuan Wang, Rui-Feng Fan, Du-Bao Yang, Dong Zhang, Lin Wang Previous studies have shown that oxidative stress-induced inhibition of autophagy plays a pivotal role in cadmium (Cd)-mediated cytotoxicity in primary rat proximal tubular (rPT) cells. The objective of this study is to explore the protective effect of puerarin (PU), a potent antioxidant, on Cd-induced autophagy inhibition and oxidative stress in rPT cells. First, Cd-induced blockage of autophagic flux in rPT cells was obviously restored by PU treatment, evidenced by immunoblot analysis of autophagy marker proteins and tandem fluorescent-tagged LC3 method. Resultantly, Cd-induced autophagosome accumulation was significantly alleviated by PU treatment. Also, Cd-induced lysosomal alkalinization and impairment of lysosomal degradation capacity were obviously recovered by PU, demonstrating that PU can restore Cd-induced lysosomal dysfunction. Moreover, Cd-induced lysosomal membrane permeabilization (LMP) was effectively blocked by PU. Cd-stimulated Nrf2 nuclear translocation and subsequent elevated expression of Nrf2-downstream targets were significantly inhibited by PU treatment. Simultaneously, Cd-elevated protein levels of antioxidant enzymes and glutathione synthesis-related proteins in rPT cells were markedly downregulated by PU treatment. In conclusion, these observations indicate that PU alleviates Cd-induced cytotoxicity in rPT cells through restoring autophagy, blocking LMP and inhibiting Nrf2 pathway, which is intimately related with its antioxidant activity.Graphical abstractGraphical abstract for this article
  • Evaluating the potential of kinase inhibitors to suppress DNA repair and
           sensitise ovarian cancer cells to PARP inhibitors
    • Abstract: Publication date: Available online 17 October 2018Source: Biochemical PharmacologyAuthor(s): Asima Mukhopadhyay, Yvette Drew, Elizabeth Matheson, Mo Salehan, Lucy Gentles, Jonathan A Pachter, Nicola J Curtin PARP inhibitors (PARPi) represent a major advance in the treatment of ovarian cancer associated with defects in homologous recombination DNA repair (HRR), primarily due to mutations in BRCA genes. Imatinib and PI3K inhibitors are reported to downregulate HRR and, in some cases, sensitise cells to PARPi.We investigated the ability of imatinib, and the PI3K inhibitors: NVP-BEZ235 and VS-5584, to downregulate HRR and sensitise paired ovarian cancer cells with mutant and reconstituted BRCA1 to the PARPi, olaparib and rucaparib. Olaparib and imatinib combinations were also measured in primary cultures of ovarian cancer.NVP-BEZ235 and imatinib reduced RAD51 levels and focus formation (an indication of HRR function), but VS-5584 did not. In colony-forming assays none of the inhibitors sensitised cells to PARPi cytotoxicity, in fact there was a mild protective effect. These conflicting data were resolved by the observation that the kinase inhibitors reduced the S-phase fraction, when HRR proteins are at their peak and cells are sensitive to PARPi cytotoxicity. In contrast, in primary cultures in 96-well plate assays, imatinib did increase olaparib-induced growth inhibition. However, in one primary culture that could be used in colony-formation cytotoxicity assays, imatinib protected from olaparib cytotoxicity.The kinase inhibitors protect from PARPi cytotoxicity by arresting cell growth, but this may be interpreted as synergy on the basis of 96-well cell growth assays. We urge caution before combining these drugs clinically. (228 words)Graphical abstractGraphical abstract for this article
  • Differences between acute and chronic stress granules, and how these
           differences may impact function in human disease
    • Abstract: Publication date: Available online 14 October 2018Source: Biochemical PharmacologyAuthor(s): Lucas C. Reineke, Joel R. Neilson Stress granules are macromolecular aggregates of mRNA and proteins assembling in response to stresses that promote the repression of protein synthesis. Most of the work characterizing stress granules has been done under acute stress conditions or during viral infection. Comparatively less work has been done to understand stress granule assembly during chronic stress, specifically regarding the composition and function of stress granules in this alternative context. Here, we describe key aspects of stress granule biology under acute stress, and how these stress granule hallmarks differ in the context of chronic stress conditions. We will provide perspective for future work aimed at further uncovering the form and function of both acute and chronic stress granules and discuss aspects of stress granule biology that have the potential to be exploited in human disease.Graphical abstractGraphical abstract for this article
  • Prenylated quinolinecarboxylic acid derivative prevents neuronal cell
           death through inhibition of MKK4
    • Abstract: Publication date: Available online 12 October 2018Source: Biochemical PharmacologyAuthor(s): Masato Ogura, Haruhisa Kikuchi, Norshalena Shakespear, Toshiyuki Suzuki, Junko Yamaki, Miwako K. Homma, Yoshiteru Oshima, Yoshimi Homma The development of neuroprotective agents is necessary for the treatment of neurodegenerative diseases. Here, we report PQA-11, a prenylated quinolinecarboxylic acid (PQA) derivative, as a potent neuroprotectant. PQA-11 inhibits glutamate-induced cell death and caspase-3 activation in hippocampal cultures, as well as inhibits N-Methyl-4-phenylpyridinium iodide- and amyloid β1-42-induced cell death in SH-SY5Y cells. PQA-11 also suppresses mitogen-activated protein kinase kinase 4 (MKK4) and c-jun N-terminal kinase (JNK) signaling activated by these neurotoxins. Quartz crystal microbalance analysis and in vitro kinase assay reveal that PQA-11 interacts with MKK4, and inhibits its sphingosine-induced activation. The administration of PQA-11 by intraperitoneal injection alleviates 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced degeneration of nigrostriatal dopaminergic neurons in mice. These results suggest that PQA-11 is a unique MKK4 inhibitor with potent neuroprotective effects in vitro and in vivo. PQA-11 may be a valuable lead for the development of novel neuroprotectants.Graphical abstractGraphical abstract for this article
  • PARP inhibition induces Akt-mediated cytoprotective effects through the
           formation of a mitochondria-targeted phospho-ATM-NEMO-Akt-mTOR signalosome
    • Abstract: Publication date: Available online 6 October 2018Source: Biochemical PharmacologyAuthor(s): Antal Tapodi, Zita Bognár, Csaba Szabo, Ferenc Gallyas, Balázs Sümegi, Enikő Hocsák PurposeThe cytoprotective effect of poly(ADP-ribose) polymerase 1 (PARP1) inhibition is well documented in various cell types subjected to oxidative stress. Previously, we have demonstrated that PARP1 inhibition activates Akt, and showed that this response plays a critical role in the maintenance of mitochondrial integrity and in cell survival. However, it has not yet been defined how nuclear PARP1 signals to cytoplasmic Akt.MethodsWRL 68, HeLa and MCF7 cells were grown in culture. Oxidative stress was induced with hydrogen peroxide. PARP was inhibited with the PARP inhibitor PJ34. ATM, mTOR- and NEMO were silenced using specific siRNAs. Cell viability assays were based on the MTT assay. PARP-ATM pulldown experiments were conducted; each protein was visualized by Western blotting. Immunoprecipitation of ATM, phospho-ATM and NEMO was performed from cytoplasmic and mitochondrial cell fractions and proteins were detected by Western blotting. In some experiments, a continually active Akt construct was introduced. Nuclear to cytoplasmic and mitochondrial translocation of phospho-Akt was visualized by confocal microscopy.ResultsHere we present evidence for a PARP1 mediated, PARylation-dependent interaction between ATM and NEMO, which is responsible for the cytoplasmic transport of phosphorylated (thus, activated) ATM kinase. In turn, the cytoplasmic p-ATM and NEMO forms complex with mTOR and Akt, yielding the phospho-ATM-NEMO-Akt-mTOR signalosome, which is responsible for the PARP-inhibition induced Akt activation. The phospho-ATM-NEMO-Akt-mTOR signalosome localizes to the mitochondria and is essential for the PARP-inhibition-mediated cytoprotective effects in oxidatively stressed cells. When the formation of the signalosome is prevented, the cytoprotective effects diminish, but cells can be rescued by constantly active Akt1, further confirming the critical role of Akt activation in cytoprotection.ConclusionsTaken together, the data presented in the current paper are consistent with the hypothesis that PARP inhibition suppresses the PARylation of ATM, which, in turn, forms an ATM-NEMO complex, which exits the nucleus, and combines in the cytosol with mTOR and Act, resulting in Act phosphorylation (i.e. activation), which, in turn, produces the cytoprotective action via the induction of Akt-mediated survival pathways. This mechanism can be important in the protective effect of PARP inhibitor in various diseases associated with oxidative stress. Moreover, disruption of the formation or action of the phospho-ATM-NEMO-Akt-mTOR signalosome may offer potential future experimental therapeutic checkpoints.Graphical abstractGraphical abstract for this article
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