Mepazine Inhibits RANK-Induced Osteoclastogenesis Independent of Its MALT1 Inhibitory Function

Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) is an intracellular cysteine protease (paracaspase) that plays an integral role in innate and adaptive immunity. The phenothiazine mepazine has been shown to inhibit the proteolytic activity of MALT1 and is frequently used to study its biological role. MALT1 has recently been suggested as a therapeutic target in rheumatoid arthritis. Here, we analyzed the effect of mepazine on the receptor activator of nuclear factor κ-B (RANK)-induced osteoclastogenesis. The treatment of mouse bone marrow precursor cells with mepazine strongly inhibited the RANK ligand (RANKL)-induced formation of osteoclasts, as well as the expression of several osteoclast markers, such as TRAP, cathepsin K, and calcitonin. However, RANKL induced osteoclastogenesis equally well in bone marrow cells derived from wild-type and Malt1 knock-out mice. Furthermore, the protective effect of mepazine was not affected by MALT1 deficiency. Additionally, the absence of MALT1 did not affect RANK-induced nuclear factor κB (NF-κB) and activator protein 1 (AP-1) activation. Overall, these studies demonstrate that MALT1 is not essential for RANK-induced osteoclastogenesis, and implicate a MALT1-independent mechanism of action of mepazine that should be taken into account in future studies using this compound.

[1]  C. Libert,et al.  Inhibition of MALT1 Decreases Neuroinflammation and Pathogenicity of Virulent Rabies Virus in Mice , 2018, Journal of Virology.

[2]  Y. Saeys,et al.  Ancient Origin of the CARD–Coiled Coil/Bcl10/MALT1-Like Paracaspase Signaling Complex Indicates Unknown Critical Functions , 2018, Front. Immunol..

[3]  R. Beyaert,et al.  Importance of Validating Antibodies and Small Compound Inhibitors Using Genetic Knockout Studies—T Cell Receptor-Induced CYLD Phosphorylation by IKKε/TBK1 as a Case Study , 2018, Front. Cell Dev. Biol..

[4]  R. Beyaert,et al.  MALT1 Controls Attenuated Rabies Virus by Inducing Early Inflammation and T Cell Activation in the Brain , 2018, Journal of Virology.

[5]  G. Bold,et al.  The T‐cell fingerprint of MALT1 paracaspase revealed by selective inhibition , 2018, Immunology and cell biology.

[6]  Chang Hoon Lee,et al.  Mucosa-associated lymphoid tissue lymphoma translocation 1 as a novel therapeutic target for rheumatoid arthritis , 2017, Scientific Reports.

[7]  L. Del Valle,et al.  A role for MALT1 activity in Kaposi's sarcoma-associated herpes virus latency and growth of primary effusion lymphoma , 2016, Leukemia.

[8]  J. Rozmus,et al.  Successful clinical treatment and functional immunological normalization of human MALT1 deficiency following hematopoietic stem cell transplantation. , 2016, Clinical immunology.

[9]  A. Bowcock,et al.  Psoriasis mutations disrupt CARD14 autoinhibition promoting BCL10-MALT1-dependent NF-κB activation. , 2016, The Biochemical journal.

[10]  N. Bidère,et al.  The paracaspase MALT1 cleaves the LUBAC subunit HOIL1 during antigen receptor signaling , 2016, Journal of Cell Science.

[11]  R. Beyaert,et al.  The paracaspase MALT1 mediates CARD14‐induced signaling in keratinocytes , 2016, EMBO reports.

[12]  Qiang Xu,et al.  MALT1 inhibitors prevent the development of DSS-induced experimental colitis in mice via inhibiting NF-κB and NLRP3 inflammasome activation , 2016, Oncotarget.

[13]  P. Hulpiau,et al.  MALT1 is not alone after all: identification of novel paracaspases , 2016, Cellular and Molecular Life Sciences.

[14]  R. Beyaert,et al.  Targeting MALT1 Proteolytic Activity in Immunity, Inflammation and Disease: Good or Bad? , 2016, Trends in molecular medicine.

[15]  R. Beyaert,et al.  MALT1 cleaves the E3 ubiquitin ligase HOIL‐1 in activated T cells, generating a dominant negative inhibitor of LUBAC‐induced NF‐κB signaling , 2016, The FEBS journal.

[16]  D. Krappmann,et al.  Combinatorial BTK and MALT1 inhibition augments killing of CD79 mutant diffuse large B cell lymphoma , 2015, Oncotarget.

[17]  R. Beyaert,et al.  MALT1 – a universal soldier: multiple strategies to ensure NF‐κB activation and target gene expression , 2015, The FEBS journal.

[18]  R. Toes,et al.  Glycosylation of immunoglobulin G determines osteoclast differentiation and bone loss , 2015, Nature Communications.

[19]  L. Xing,et al.  NF-κB-Mediated Regulation of Osteoclastogenesis , 2015, Endocrinology and metabolism.

[20]  E. Bowman,et al.  Crosstalk among IL-23 and DNAX Activating Protein of 12 kDa–Dependent Pathways Promotes Osteoclastogenesis , 2014, The Journal of Immunology.

[21]  M. Prinz,et al.  Pharmacological inhibition of MALT1 protease activity protects mice in a mouse model of multiple sclerosis , 2014, Journal of Neuroinflammation.

[22]  R. Holt,et al.  Combined immunodeficiency associated with homozygous MALT1 mutations. , 2014, The Journal of allergy and clinical immunology.

[23]  J. Ruland,et al.  Structural analysis of phenothiazine derivatives as allosteric inhibitors of the MALT1 paracaspase. , 2013, Angewandte Chemie.

[24]  M. R. Islam,et al.  CMV promoter is repressed by p53 and activated by JNK pathway. , 2013, Plasmid.

[25]  G. Lenz,et al.  Pharmacologic inhibition of MALT1 protease by phenothiazines as a therapeutic approach for the treatment of aggressive ABC-DLBCL. , 2012, Cancer cell.

[26]  H. Takayanagi,et al.  New insights into osteoclastogenic signaling mechanisms , 2012, Trends in Endocrinology & Metabolism.

[27]  Agata Jaszczyszyn,et al.  Chemical structure of phenothiazines and their biological activity , 2012, Pharmacological reports : PR.

[28]  J. Trowsdale,et al.  OSCAR is a collagen receptor that costimulates osteoclastogenesis in DAP12-deficient humans and mice. , 2011, The Journal of clinical investigation.

[29]  M. Kool,et al.  A20 (TNFAIP3) deficiency in myeloid cells triggers erosive polyarthritis resembling rheumatoid arthritis , 2011, Nature Genetics.

[30]  G. Lenz,et al.  Malt1-dependent RelB cleavage promotes canonical NF-κB activation in lymphocytes and lymphoma cell lines , 2011, Proceedings of the National Academy of Sciences.

[31]  Andreas Radbruch,et al.  Dephosphorylation of Bcl‐10 by calcineurin is essential for canonical NF‐κB activation in Th cells , 2011, European journal of immunology.

[32]  K. Pluta,et al.  Recent progress in biological activities of synthesized phenothiazines. , 2011, European journal of medicinal chemistry.

[33]  T. Grundström,et al.  CaMKII targets Bcl10 in T-cell receptor induced activation of NF-κB. , 2011, Molecular immunology.

[34]  K. Gevaert,et al.  T‐cell receptor‐induced JNK activation requires proteolytic inactivation of CYLD by MALT1 , 2011, The EMBO journal.

[35]  M. Karsdal,et al.  Osteoclast activity and subtypes as a function of physiology and pathology--implications for future treatments of osteoporosis. , 2011, Endocrine reviews.

[36]  A. Hess,et al.  Increased bone density and resistance to ovariectomy-induced bone loss in FoxP3-transgenic mice based on impaired osteoclast differentiation. , 2010, Arthritis and rheumatism.

[37]  C. Chu,et al.  The expanding roles of ITAM adapters FcRγ and DAP12 in myeloid cells , 2009, Immunological reviews.

[38]  Shao-Cong Sun,et al.  Deubiquitinating enzyme CYLD negatively regulates RANK signaling and osteoclastogenesis in mice. , 2008, The Journal of clinical investigation.

[39]  W. Zou,et al.  RelB is the NF-κB subunit downstream of NIK responsible for osteoclast differentiation , 2008, Proceedings of the National Academy of Sciences.

[40]  P. Marynen,et al.  T cell antigen receptor stimulation induces MALT1 paracaspase–mediated cleavage of the NF-κB inhibitor A20 , 2008, Nature Immunology.

[41]  R. Xavier,et al.  Bcl10 is phosphorylated on Ser138 by Ca2+/calmodulin-dependent protein kinase II. , 2007, Molecular immunology.

[42]  L. Piccio,et al.  Cutting Edge: TREM-2 Attenuates Macrophage Activation1 , 2006, The Journal of Immunology.

[43]  M. Daly,et al.  Ca2+/Calmodulin-Dependent Protein Kinase II Is a Modulator of CARMA1-Mediated NF-κB Activation , 2006, Molecular and Cellular Biology.

[44]  E. Wagner,et al.  Fos/AP‐1 proteins in bone and the immune system , 2005, Immunological reviews.

[45]  A. Kudo,et al.  Self‐Assembled RANK Induces Osteoclastogenesis Ligand‐Independently , 2005, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[46]  M. Santis,et al.  Rationale for T Cell Inhibition by Cyclosporin A in Major Autoimmune Diseases , 2005, Annals of the New York Academy of Sciences.

[47]  G. Weber,et al.  Synergistic activation of the CMV promoter by NF-κB P50 and PKG , 2004 .

[48]  Jürgen Ruland,et al.  Differential requirement for Malt1 in T and B cell antigen receptor signaling. , 2003, Immunity.

[49]  E. Wagner,et al.  Signalling in osteoclasts and the role of Fos/AP1 proteins , 2003, Annals of the rheumatic diseases.

[50]  J. McDonald,et al.  The role of calmodulin in the regulation of osteoclastogenesis. , 2003, Endocrinology.

[51]  David L. Lacey,et al.  Osteoclast differentiation and activation , 2003, Nature.

[52]  Hiroshi Takayanagi,et al.  Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal differentiation of osteoclasts. , 2002, Developmental cell.

[53]  Matthias Brüstle,et al.  Chemtool - Moleküle zeichnen mit dem Pinguin , 2001 .

[54]  M. Bezerra,et al.  Effects of chlorpromazine on alveolar bone loss in experimental periodontal disease in rats. , 2000, European journal of oral sciences.

[55]  W. Dougall,et al.  RANK is essential for osteoclast and lymph node development. , 1999, Genes & development.

[56]  R. Dubose,et al.  A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function , 1997, Nature.

[57]  T. J. Hall,et al.  Phenothiazines are potent inhibitors of osteoclastic bone resorption. , 1996, General pharmacology.

[58]  T. J. Hall,et al.  Promethazine inhibits osteoclastic bone resorption In vitro , 1994, Calcified Tissue International.

[59]  Sheila J. Jones,et al.  Osteoclast-like cell formation and its regulation by osteotropic hormones in mouse bone marrow cultures. , 1988, Endocrinology.

[60]  N. Kurihara,et al.  Effect of chlorpromazine on PZ-peptidase and several other peptidase activities in cloned osteoblastic cells (MC3T3-E1). , 1987, Biochemical pharmacology.

[61]  M. Kumegawa,et al.  Inhibitory effect of phenothiazine derivatives on bone in vivo and osteoblastic cells in vitro. , 1985, Biochemical pharmacology.

[62]  B. Weiss,et al.  Interaction of drugs with calmodulin. Biochemical, pharmacological and clinical implications. , 1982, Biochemical pharmacology.

[63]  P. Goldhaber,et al.  Effect of Promethazine Hydrochloride on Bone Resorption in Tissue Culture 1 , 1982, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[64]  D. Klein,et al.  Mepazine (pacatal): Clinical trial with placebo control and psychological study , 1960, Psychopharmacologia.

[65]  H. Bowes.  The ataractic drugs: the present position of chlorpromazine, frenquel, pacatal, and reserpine in the psychiatric hospital. , 1956 .

[66]  A. Togari,et al.  Inhibitory effect of chlorpromazine on RANKL-induced osteoclastogenesis in mouse bone marrow cells. , 2011, Journal of pharmacological sciences.