The Experts below are selected from a list of 639 Experts worldwide ranked by ideXlab platform
Rudi Beyaert - One of the best experts on this subject based on the ideXlab platform.
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Mepazine Inhibits RANK-Induced Osteoclastogenesis Independent of Its MALT1 Inhibitory Function
Molecules, 2018Co-Authors: Laura Meloni, Jens Staal, Yasmine Driege, Inna S Afonina, Marja Kreike, Lynn Verstrepen, Rudi BeyaertAbstract: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.
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card14 mediated activation of Paracaspase malt1 in keratinocytes implications for psoriasis
Journal of Investigative Dermatology, 2017Co-Authors: Elien Van Nuffel, Rudi Beyaert, Inna S Afonina, Anja Schmitt, Klaus Schulzeosthoff, Stephan HailfingerAbstract:Mutations in caspase recruitment domain-containing protein 14 (CARD14) have been linked to susceptibility to psoriasis. CARD14 is an intracellular scaffold protein that regulates proinflammatory gene expression. Recent studies have offered novel insights into the mechanisms of CARD14-mediated signaling in keratinocytes and the molecular impact of psoriasis-associated CARD14 mutations. CARD14 forms a signaling complex with BCL10 and the Paracaspase MALT1, and this process is enhanced upon pathogenic CARD14 mutation, culminating in the activation of MALT1 protease activity and psoriasis-associated gene expression. This review summarizes the current knowledge of CARD14/MALT1-mediated signaling in keratinocytes and its therapeutic implications in psoriasis.
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02 08 the Paracaspase malt1 plays a central role in the pathogenesis of rheumatoid arthritis
Annals of the Rheumatic Diseases, 2017Co-Authors: Elisabeth Gilis, Jens Staal, Rudi Beyaert, Dirk ElewautAbstract:Background One of the hallmarks of many inflammatory arthritides is their strong linkage with MHC-signalling, which is mirrored by the marked role for adaptive immunity. Accordingly, rheumatoid arthritis (RA) is characterized by the activation of auto-reactive T-cells and the development of auto-antibodies. T-cells may additionally respond to non-TCR mediated signals, which are essential in driving their effector functions. Pathways leading to the modulation of both innate and adaptive signals are therefore of marked interest to study in arthritic diseases. Objectives The Paracaspase MALT1 is a key player in the activation and proliferation of immune and non-immune cells. These cells include the lymphoid, myeloid and mast cells, indicating MALT19s crucial role in both innate and adaptive signaling (1). Therefore, MALT1 is regarded a promising target for the treatment of autoimmune diseases and defining its role in the pathogenesis of inflammatory arthritis is a critical first step. Methods To unravel MALT19s role in inflammatory arthritis, we initially assessed MALT1-activation in mice that were challenged with collagen-induced arthritis (CIA), the prototype model for antigen-induced RA. We then addressed the role of MALT1 in the pathogenesis of inflammatory arthritis by challenging MALT1-deficient mice to distinct models of arthritis (CIA and CAIA) or by backcrossing MALT1-deficient mice to TNF DARE mice, representing an SpA-like model. Additionally, CIA was induced in CD4-specific MALT1-deficient mice to determine the importance of MALT1 in T-cells. Results We provide evidence that MALT1 plays a crucial role in the pathogenesis of RA as MALT1-deficent mice were completely protected against CIA. This complete protection was additionally observed in CD4-specific MALT1-deficient mice, indicating that the selective ablation of MALT1 in CD4-positive cells is sufficient for the observed resistance against CIA. CAIA on the other hand, which is a T- and B-cell independent model of RA, did not depend on the presence of MALT1, since both MALT1 +/+ and MALT1 -/- mice showed comparable symptoms of RA. Interestingly, TNF DARE mice that were deficient for MALT1 also showed a reduced enthesitis and ileitis phenotype, although TNF-concentration in the serum of these mice was higher compared to MALT1 +/+ xTNF DARE mice. Conclusions Overall, our data highlight that MALT1 plays a crucial role in the pathogenesis of inflammatory arthritis and represents an interesting candidate to target therapeutically. References Thome M. Multifunctional roles for MALT1 in T-cell activation. Nat Rev Immunol 2008; 8 (7): 495–500. Acknowledgements We thank Chris Vercruysse (Department of Basic Medical Sciences, University of Ghent, Belgium) for exerting the three-point-bending tests of the femurs and the lab of Prof. Dr. Luc Van Hoorebeke (Department of Physics and Astronomy) for the use of the μCT-scanner. Disclosure of Interest None declared
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the Paracaspase malt1 mediates card14 induced signaling in keratinocytes
EMBO Reports, 2016Co-Authors: Inna S Afonina, Jens Staal, Yasmine Driege, Elien Van Nuffel, Griet Baudelet, Marja Kreike, Rudi BeyaertAbstract:Mutations in CARD14 have recently been linked to psoriasis susceptibility. CARD14 is an epidermal regulator of NF‐κB activation. However, the ability of CARD14 to activate other signaling pathways as well as the biochemical mechanisms that mediate and regulate its function remain to be determined. Here, we report that in addition to NF‐κB signaling, CARD14 activates p38 and JNK MAP kinase pathways, all of which are dependent on the Paracaspase MALT1. Mechanistically, we demonstrate that CARD14 physically interacts with Paracaspase MALT1 and activates MALT1 proteolytic activity and inflammatory gene expression, which are enhanced by psoriasis‐associated CARD14 mutations. Moreover, we show that MALT1 deficiency or pharmacological inhibition of MALT1 catalytic activity inhibits pathogenic mutant CARD14‐induced cytokine and chemokine expression in human primary keratinocytes. Collectively, our findings demonstrate a novel role for MALT1 in CARD14‐induced signaling and indicate MALT1 as a valuable therapeutic target in psoriasis. ![][1] This study shows that the Paracaspase MALT1 is indispensable for CARD14‐induced NF‐κB and MAP kinase signaling. Pharmacological inhibition of MALT1 prevents pro‐inflammatory gene expression in primary keratinocytes induced by psoriasis‐associated mutation of CARD14. EMBO Reports (2016) 17: 914–927 [1]: /embed/graphic-1.gif
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The Paracaspase MALT1 mediates CARD14‐induced signaling in keratinocytes
EMBO Reports, 2016Co-Authors: Inna S Afonina, Jens Staal, Yasmine Driege, Elien Van Nuffel, Griet Baudelet, Marja Kreike, Rudi BeyaertAbstract:Mutations in CARD14 have recently been linked to psoriasis susceptibility. CARD14 is an epidermal regulator of NF‐κB activation. However, the ability of CARD14 to activate other signaling pathways as well as the biochemical mechanisms that mediate and regulate its function remain to be determined. Here, we report that in addition to NF‐κB signaling, CARD14 activates p38 and JNK MAP kinase pathways, all of which are dependent on the Paracaspase MALT1. Mechanistically, we demonstrate that CARD14 physically interacts with Paracaspase MALT1 and activates MALT1 proteolytic activity and inflammatory gene expression, which are enhanced by psoriasis‐associated CARD14 mutations. Moreover, we show that MALT1 deficiency or pharmacological inhibition of MALT1 catalytic activity inhibits pathogenic mutant CARD14‐induced cytokine and chemokine expression in human primary keratinocytes. Collectively, our findings demonstrate a novel role for MALT1 in CARD14‐induced signaling and indicate MALT1 as a valuable therapeutic target in psoriasis. ![][1] This study shows that the Paracaspase MALT1 is indispensable for CARD14‐induced NF‐κB and MAP kinase signaling. Pharmacological inhibition of MALT1 prevents pro‐inflammatory gene expression in primary keratinocytes induced by psoriasis‐associated mutation of CARD14. EMBO Reports (2016) 17: 914–927 [1]: /embed/graphic-1.gif
Jens Staal - One of the best experts on this subject based on the ideXlab platform.
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Mepazine Inhibits RANK-Induced Osteoclastogenesis Independent of Its MALT1 Inhibitory Function
Molecules, 2018Co-Authors: Laura Meloni, Jens Staal, Yasmine Driege, Inna S Afonina, Marja Kreike, Lynn Verstrepen, Rudi BeyaertAbstract: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.
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ancient origin of the card coiled coil bcl10 malt1 like Paracaspase signaling complex indicates unknown critical functions
Frontiers in Immunology, 2018Co-Authors: Jens Staal, Yasmine Driege, Paco Hulpiau, Mira Haegman, Alice Borghi, Laurens Lievens, Srividhya Sundararaman, Amanda Goncalves, Ineke Dhondt, Jorge H PinzonAbstract:The CARD-coiled coil (CC)/Bcl10/MALT1-like Paracaspase (CBM) signaling complexes composed of a CARD-CC family member (CARD-9, -10, -11, or -14), Bcl10, and the type 1 Paracaspase MALT1 (PCASP1) play a pivotal role in immunity, inflammation, and cancer. Targeting MALT1 proteolytic activity is of potential therapeutic interest. However, little is known about the evolutionary origin and the original functions of the CBM complex. Type 1 Paracaspases originated before the last common ancestor of planulozoa (bilaterians and cnidarians). Notably in bilaterians, Ecdysozoa (e.g., nematodes and insects) lacks Bcl10, whereas other lineages have a Bcl10 homolog. A survey of invertebrate CARD-CC homologs revealed such homologs only in species with Bcl10, indicating an ancient common origin of the entire CBM complex. Furthermore, vertebrate-like Syk/Zap70 tyrosine kinase homologs with the ITAM-binding SH2 domain were only found in invertebrate organisms with CARD-CC/Bcl10, indicating that this pathway might be related to the original function of the CBM complex. Moreover, the type 1 Paracaspase sequences from invertebrate organisms that have CARD-CC/Bcl10 are more similar to vertebrate Paracaspases. Functional analysis of protein-protein interactions, NF-kappa B signaling, and CYLD cleavage for selected invertebrate type 1 Paracaspase and Bcl10 homologs supports this scenario and indicates an ancient origin of the CARD-CC/Bcl10/Paracaspase signaling complex. By contrast, many of the known MALT1-associated activities evolved fairly recently, indicating that unknown functions are at the basis of the protein conservation. As a proof-of-concept, we provide initial evidence for a CBM- and NF-kappa B-independent neuronal function of the Caenorhabditis elegans type 1 Paracaspase malt-1. In conclusion, this study shows how evolutionary insights may point at alternative functions of MALT1.
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Ancient Origin of the CARD-Coiled Coil/Bcl10/MALT1-Like Paracaspase Signaling Complex Indicates Unknown Critical Functions.
Frontiers in Immunology, 2018Co-Authors: Jens Staal, Yasmine Driege, Paco Hulpiau, Mira Haegman, Alice Borghi, Laurens Lievens, Srividhya Sundararaman, Amanda Goncalves, Ineke DhondtAbstract:The CARD-coiled coil (CC)/Bcl10/MALT1-like Paracaspase (CBM) signaling complexes composed of a CARD-CC family member (CARD-9, -10, -11, or -14), Bcl10, and the type 1 Paracaspase MALT1 (PCASP1) play a pivotal role in immunity, inflammation, and cancer. Targeting MALT1 proteolytic activity is of potential therapeutic interest. However, little is known about the evolutionary origin and the original functions of the CBM complex. Type 1 Paracaspases originated before the last common ancestor of planulozoa (bilaterians and cnidarians). Notably in bilaterians, Ecdysozoa (e.g., nematodes and insects) lacks Bcl10, whereas other lineages have a Bcl10 homolog. A survey of invertebrate CARD-CC homologs revealed such homologs only in species with Bcl10, indicating an ancient common origin of the entire CBM complex. Furthermore, vertebrate-like Syk/Zap70 tyrosine kinase homologs with the ITAM-binding SH2 domain were only found in invertebrate organisms with CARD-CC/Bcl10, indicating that this pathway might be related to the original function of the CBM complex. Moreover, the type 1 Paracaspase sequences from invertebrate organisms that have CARD-CC/Bcl10 are more similar to vertebrate Paracaspases. Functional analysis of protein-protein interactions, NF-kappa B signaling, and CYLD cleavage for selected invertebrate type 1 Paracaspase and Bcl10 homologs supports this scenario and indicates an ancient origin of the CARD-CC/Bcl10/Paracaspase signaling complex. By contrast, many of the known MALT1-associated activities evolved fairly recently, indicating that unknown functions are at the basis of the protein conservation. As a proof-of-concept, we provide initial evidence for a CBM- and NF-kappa B-independent neuronal function of the Caenorhabditis elegans type 1 Paracaspase malt-1. In conclusion, this study shows how evolutionary insights may point at alternative functions of MALT1.
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GC content of Early Metazoan genes and its impact on gene expression levels in mammalian cell lines
Genome Biology and Evolution, 2018Co-Authors: Jens Staal, Yasmine Driege, Paco Hulpiau, Evi De Keuckelaere, Kai Kamm, Tom Deroo, Ellen Sanders, Katrien Staes, Yvan SaeysAbstract:With the genomes available for many animal clades, including the early-branching metazoans, one can readily study the functional conservation of genes across a diversity of animal lineages. Ectopic expression of an animal protein in, for instance, a mammalian cell line is a generally used strategy in structure–function analysis. However, this might turn out to be problematic in case of distantly related species. Here we analyzed the GC content of the coding sequences of basal animals and show its impact on gene expression levels in human cell lines, and, importantly, how this expression efficiency can be improved. Optimization of the GC3 content in the coding sequences of cadherin, alpha-catenin, and Paracaspase of Trichoplax adhaerens dramatically increased the expression of these basal animal genes in human cell lines.
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02 08 the Paracaspase malt1 plays a central role in the pathogenesis of rheumatoid arthritis
Annals of the Rheumatic Diseases, 2017Co-Authors: Elisabeth Gilis, Jens Staal, Rudi Beyaert, Dirk ElewautAbstract:Background One of the hallmarks of many inflammatory arthritides is their strong linkage with MHC-signalling, which is mirrored by the marked role for adaptive immunity. Accordingly, rheumatoid arthritis (RA) is characterized by the activation of auto-reactive T-cells and the development of auto-antibodies. T-cells may additionally respond to non-TCR mediated signals, which are essential in driving their effector functions. Pathways leading to the modulation of both innate and adaptive signals are therefore of marked interest to study in arthritic diseases. Objectives The Paracaspase MALT1 is a key player in the activation and proliferation of immune and non-immune cells. These cells include the lymphoid, myeloid and mast cells, indicating MALT19s crucial role in both innate and adaptive signaling (1). Therefore, MALT1 is regarded a promising target for the treatment of autoimmune diseases and defining its role in the pathogenesis of inflammatory arthritis is a critical first step. Methods To unravel MALT19s role in inflammatory arthritis, we initially assessed MALT1-activation in mice that were challenged with collagen-induced arthritis (CIA), the prototype model for antigen-induced RA. We then addressed the role of MALT1 in the pathogenesis of inflammatory arthritis by challenging MALT1-deficient mice to distinct models of arthritis (CIA and CAIA) or by backcrossing MALT1-deficient mice to TNF DARE mice, representing an SpA-like model. Additionally, CIA was induced in CD4-specific MALT1-deficient mice to determine the importance of MALT1 in T-cells. Results We provide evidence that MALT1 plays a crucial role in the pathogenesis of RA as MALT1-deficent mice were completely protected against CIA. This complete protection was additionally observed in CD4-specific MALT1-deficient mice, indicating that the selective ablation of MALT1 in CD4-positive cells is sufficient for the observed resistance against CIA. CAIA on the other hand, which is a T- and B-cell independent model of RA, did not depend on the presence of MALT1, since both MALT1 +/+ and MALT1 -/- mice showed comparable symptoms of RA. Interestingly, TNF DARE mice that were deficient for MALT1 also showed a reduced enthesitis and ileitis phenotype, although TNF-concentration in the serum of these mice was higher compared to MALT1 +/+ xTNF DARE mice. Conclusions Overall, our data highlight that MALT1 plays a crucial role in the pathogenesis of inflammatory arthritis and represents an interesting candidate to target therapeutically. References Thome M. Multifunctional roles for MALT1 in T-cell activation. Nat Rev Immunol 2008; 8 (7): 495–500. Acknowledgements We thank Chris Vercruysse (Department of Basic Medical Sciences, University of Ghent, Belgium) for exerting the three-point-bending tests of the femurs and the lab of Prof. Dr. Luc Van Hoorebeke (Department of Physics and Astronomy) for the use of the μCT-scanner. Disclosure of Interest None declared
Changying Jiang - One of the best experts on this subject based on the ideXlab platform.
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targeting the Paracaspase malt1 a potential therapy to overcome ibrutinib resistance in relapsed refractory mantle cell lymphoma patients
Blood, 2017Co-Authors: Changying Jiang, Shengjian Huang, Carrie J Li, Jeff F Wang, Elyse R Lopez, Hui Zhang, Elizabeth Lorence, Maria Merolle, Swathi Balaji, Makhdum AhmedAbstract:Mantle cell lymphoma (MCL) is an aggressive B cell malignancy that is not yet curable. Ibrutinib was approved in 2013 for the treatment of relapsed/refractory MCL patients; however, ibrutinib resistance inevitably develops. Therefore, there is an urgent unmet need to overcome ibrutinib resistance and to study alternative treatment options. Constitutive NF-κB activation is the hallmark of MCL. Indeed, next generation sequencing analysis of 110 MCL patient samples revealed that genes belonging to the NF-κB signaling pathway had a high mutation rate (28.8%), indicating a significant contribution of NF-κB signaling to MCL malignancy. Mucosa-associated lymphoid tissue transformation protein (MALT1) plays a crucial role in NF-κB signaling. MALT1 is a unique Paracaspase within the human genome, and the proteolytic activity of MALT1 has been found to be constitutively active in many MCL samples, suggesting that MALT1 may be a potential therapeutic target without significant off-target side effects. MI-2 is a specific inhibitor of MALT1 and its safety and efficacy are currently being examined in a clinical trial for the ABC subtype of diffuse large B cell lymphoma. In addition, animal models treated with MI-2 did not have any detectable physiological, histological or biochemical signs of toxicity. However, whether MALT1 activity contributes to ibrutinib resistance and whether targeting MALT1 can overcome ibrutinib resistance in relapsed/refractory MCL patients remain unclear. In this study, we found that both canonical and non-canonical NF-κB signaling is activated in ibrutinib-resistant MCL cells, which correlates with constitutive MALT1 activity. Interestingly, we found that MALT1 is highly mutated in four clusters including the death domain, TRAF6 binding site, Caspase like domain, and IKKγ binding site. Occurrence of L79P, K80R, E319D, L445P and N446S strongly correlated with ibrutinib resistance and disease progression. Treatment with MI-2 at nanomolar levels significantly reduced cell viability and induced apoptosis in several MCL cell lines. MI-2 treatment inhibited NF-κB activation, IL-6 production and downstream STAT3 activation. Combination of MI-2 and ibrutinib resulted in synergistic growth inhibition in both ibrutinib-resistant MCL cell lines and primary MCL cells. RPPA analysis showed that treatment with the MI-2 and ibrutinib combination resulted in marked changes in the cellular protein profiles, suggesting that targeting MALT1 catalytic activity in MCL is a promising therapy to overcome ibrutinib resistance in relapsed/refractory MCL patients. Furthermore, MI-2 treatment greatly inhibited the in vitro cell migration of the majority of examined MCL cell lines as well as cell adhesion to the extracellular matrix, including integrin and laminin; therefore, treatment with MI-2 may induce lymphocytosis (i.e., compartmental shift) in patients and mouse xenograft models. Altogether, our data suggest that targeting MALT1 is a very promising therapy for treating relapsed/refractory MCL patients. These studies support the development of preclinical and clinical studies to overcome ibrutinib resistance by targeting MALT1. Disclosures Wang: Proteolix: Honoraria, Research Funding; Pharmacyclics: Research Funding; Onyx: Research Funding; Kite Pharma: Research Funding; Juno Therapeutics: Research Funding; Janssen: Consultancy, Honoraria, Membership on an entity9s Board of Directors or advisory committees, Research Funding; Dava Oncology: Honoraria; Celgene: Honoraria, Research Funding; BeiGene: Research Funding; Asana Biosciences: Research Funding; Acerta Pharma: Consultancy, Research Funding.
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Targeting the Paracaspase MALT1: A Potential Therapy to Overcome Ibrutinib Resistance in Relapsed/Refractory Mantle Cell Lymphoma Patients
Blood, 2017Co-Authors: Changying Jiang, Shengjian Huang, Carrie J Li, Jeff F Wang, Elyse R Lopez, Hui Zhang, Elizabeth Lorence, Maria MerolleAbstract:Mantle cell lymphoma (MCL) is an aggressive B cell malignancy that is not yet curable. Ibrutinib was approved in 2013 for the treatment of relapsed/refractory MCL patients; however, ibrutinib resistance inevitably develops. Therefore, there is an urgent unmet need to overcome ibrutinib resistance and to study alternative treatment options. Constitutive NF-κB activation is the hallmark of MCL. Indeed, next generation sequencing analysis of 110 MCL patient samples revealed that genes belonging to the NF-κB signaling pathway had a high mutation rate (28.8%), indicating a significant contribution of NF-κB signaling to MCL malignancy. Mucosa-associated lymphoid tissue transformation protein (MALT1) plays a crucial role in NF-κB signaling. MALT1 is a unique Paracaspase within the human genome, and the proteolytic activity of MALT1 has been found to be constitutively active in many MCL samples, suggesting that MALT1 may be a potential therapeutic target without significant off-target side effects. MI-2 is a specific inhibitor of MALT1 and its safety and efficacy are currently being examined in a clinical trial for the ABC subtype of diffuse large B cell lymphoma. In addition, animal models treated with MI-2 did not have any detectable physiological, histological or biochemical signs of toxicity. However, whether MALT1 activity contributes to ibrutinib resistance and whether targeting MALT1 can overcome ibrutinib resistance in relapsed/refractory MCL patients remain unclear. In this study, we found that both canonical and non-canonical NF-κB signaling is activated in ibrutinib-resistant MCL cells, which correlates with constitutive MALT1 activity. Interestingly, we found that MALT1 is highly mutated in four clusters including the death domain, TRAF6 binding site, Caspase like domain, and IKKγ binding site. Occurrence of L79P, K80R, E319D, L445P and N446S strongly correlated with ibrutinib resistance and disease progression. Treatment with MI-2 at nanomolar levels significantly reduced cell viability and induced apoptosis in several MCL cell lines. MI-2 treatment inhibited NF-κB activation, IL-6 production and downstream STAT3 activation. Combination of MI-2 and ibrutinib resulted in synergistic growth inhibition in both ibrutinib-resistant MCL cell lines and primary MCL cells. RPPA analysis showed that treatment with the MI-2 and ibrutinib combination resulted in marked changes in the cellular protein profiles, suggesting that targeting MALT1 catalytic activity in MCL is a promising therapy to overcome ibrutinib resistance in relapsed/refractory MCL patients. Furthermore, MI-2 treatment greatly inhibited the in vitro cell migration of the majority of examined MCL cell lines as well as cell adhesion to the extracellular matrix, including integrin and laminin; therefore, treatment with MI-2 may induce lymphocytosis (i.e., compartmental shift) in patients and mouse xenograft models. Altogether, our data suggest that targeting MALT1 is a very promising therapy for treating relapsed/refractory MCL patients. These studies support the development of preclinical and clinical studies to overcome ibrutinib resistance by targeting MALT1. Disclosures Wang: Proteolix: Honoraria, Research Funding; Pharmacyclics: Research Funding; Onyx: Research Funding; Kite Pharma: Research Funding; Juno Therapeutics: Research Funding; Janssen: Consultancy, Honoraria, Membership on an entity9s Board of Directors or advisory committees, Research Funding; Dava Oncology: Honoraria; Celgene: Honoraria, Research Funding; BeiGene: Research Funding; Asana Biosciences: Research Funding; Acerta Pharma: Consultancy, Research Funding.
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abstract 4099 targeting the Paracaspase malt1 a potential therapy to overcome ibrutinib resistance in relapsed refractory mcl patients
Cancer Research, 2017Co-Authors: Changying Jiang, Shengjian Huang, Michael Wang, Liang ZhangAbstract:Mantle cell lymphoma (MCL) is an aggressive B cell malignancy that is not yet curable. Ibrutinib was FDA-approved in 2013 to treat relapsed/refractory MCL; however, ibrutinib resistance inevitably develops. Once patients relapse after ibrutinib treatment, the 1-year survival rate is only 22%; therefore, there is an urgent unmet need to overcome ibrutinib resistance and to study alternative treatment options. Constitutive NF-κB activation is the hallmark of MCL. Indeed, next generation sequencing analysis of 110 MCL patient samples revealed that genes in the NF-κB signaling pathway had the highest mutation rates (28.8%), indicating the significant contribution of NF-κB signaling to MCL malignancy. Mucosa-associated lymphoid tissue transformation protein (MALT1) plays a crucial role in NF-κB signaling. MALT1 is a unique Paracaspase within the human genome, and the proteolytic activity of MALT1 has been found to be constitutively active in many MCL samples, suggesting MALT1 may be a potential therapeutic target without significant off-target side effects. MI-2 is a specific inhibitor of MALT1 and its efficacy and safety are currently being evaluated in a clinical trial with ABC-type diffuse large B cell lymphoma patients. Mice treated with MI-2 did not have detectable physiological, histological or biochemical signs of toxicity. However, whether MALT1 activity contributes to ibrutinib resistance and whether targeting MALT1 can overcome ibrutinib resistance in relapsed/refractory MCL patients remain unclear. In this study, we found that both canonical and non-canonical NF-κB signaling is activated in ibrutinib-resistant MCL cells, which correlates with constitutive MALT1 activity. Interestingly, we found that MALT1 is highly mutated in four clusters, including the death domain, TRAF6-binding site, Caspase-like domain, and IKKγ-binding site in MCL samples. Occurrence of L79P, K80R, E319D, L445P and N446S was also highly correlated with ibrutinib resistance and disease progression, which requires more detailed investigation. Treatment with MI-2 significantly reduced cell viability in several MCL cell lines with nanomolar activity. MI-2 treatment inhibited NF-κB activation, IL-6 production and its downstream STAT3 activation. Combining MI-2 with ibrutinib resulted in synergistic growth inhibition in both ibrutinib-resistant MCL cell lines and primary MCL cells. These findings suggest that targeting MALT1 catalytic activity in MCL is a promising therapy to overcome ibrutinib resistance in relapsed/refractory MCL patients. The effect of MI-2 in in vivo PDX models is currently under investigation. This work and follow-up in vitro and in vivo studies will provide strong evidence that targeting MALT1 with MI-2 may be an effective novel therapeutic approach to overcome ibrutinib resistance. Citation Format: Changying Jiang, Shengjian Huang, Xin Lin, Michael Wang, Liang Zhang. Targeting the Paracaspase MALT1: A potential therapy to overcome ibrutinib resistance in relapsed/refractory MCL patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4099. doi:10.1158/1538-7445.AM2017-4099
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Abstract 4099: Targeting the Paracaspase MALT1: A potential therapy to overcome ibrutinib resistance in relapsed/refractory MCL patients
Cancer Research, 2017Co-Authors: Changying Jiang, Shengjian Huang, Michael Wang, Liang ZhangAbstract:Mantle cell lymphoma (MCL) is an aggressive B cell malignancy that is not yet curable. Ibrutinib was FDA-approved in 2013 to treat relapsed/refractory MCL; however, ibrutinib resistance inevitably develops. Once patients relapse after ibrutinib treatment, the 1-year survival rate is only 22%; therefore, there is an urgent unmet need to overcome ibrutinib resistance and to study alternative treatment options. Constitutive NF-κB activation is the hallmark of MCL. Indeed, next generation sequencing analysis of 110 MCL patient samples revealed that genes in the NF-κB signaling pathway had the highest mutation rates (28.8%), indicating the significant contribution of NF-κB signaling to MCL malignancy. Mucosa-associated lymphoid tissue transformation protein (MALT1) plays a crucial role in NF-κB signaling. MALT1 is a unique Paracaspase within the human genome, and the proteolytic activity of MALT1 has been found to be constitutively active in many MCL samples, suggesting MALT1 may be a potential therapeutic target without significant off-target side effects. MI-2 is a specific inhibitor of MALT1 and its efficacy and safety are currently being evaluated in a clinical trial with ABC-type diffuse large B cell lymphoma patients. Mice treated with MI-2 did not have detectable physiological, histological or biochemical signs of toxicity. However, whether MALT1 activity contributes to ibrutinib resistance and whether targeting MALT1 can overcome ibrutinib resistance in relapsed/refractory MCL patients remain unclear. In this study, we found that both canonical and non-canonical NF-κB signaling is activated in ibrutinib-resistant MCL cells, which correlates with constitutive MALT1 activity. Interestingly, we found that MALT1 is highly mutated in four clusters, including the death domain, TRAF6-binding site, Caspase-like domain, and IKKγ-binding site in MCL samples. Occurrence of L79P, K80R, E319D, L445P and N446S was also highly correlated with ibrutinib resistance and disease progression, which requires more detailed investigation. Treatment with MI-2 significantly reduced cell viability in several MCL cell lines with nanomolar activity. MI-2 treatment inhibited NF-κB activation, IL-6 production and its downstream STAT3 activation. Combining MI-2 with ibrutinib resulted in synergistic growth inhibition in both ibrutinib-resistant MCL cell lines and primary MCL cells. These findings suggest that targeting MALT1 catalytic activity in MCL is a promising therapy to overcome ibrutinib resistance in relapsed/refractory MCL patients. The effect of MI-2 in in vivo PDX models is currently under investigation. This work and follow-up in vitro and in vivo studies will provide strong evidence that targeting MALT1 with MI-2 may be an effective novel therapeutic approach to overcome ibrutinib resistance. Citation Format: Changying Jiang, Shengjian Huang, Xin Lin, Michael Wang, Liang Zhang. Targeting the Paracaspase MALT1: A potential therapy to overcome ibrutinib resistance in relapsed/refractory MCL patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4099. doi:10.1158/1538-7445.AM2017-4099
Yasmine Driege - One of the best experts on this subject based on the ideXlab platform.
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Mepazine Inhibits RANK-Induced Osteoclastogenesis Independent of Its MALT1 Inhibitory Function
Molecules, 2018Co-Authors: Laura Meloni, Jens Staal, Yasmine Driege, Inna S Afonina, Marja Kreike, Lynn Verstrepen, Rudi BeyaertAbstract: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.
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ancient origin of the card coiled coil bcl10 malt1 like Paracaspase signaling complex indicates unknown critical functions
Frontiers in Immunology, 2018Co-Authors: Jens Staal, Yasmine Driege, Paco Hulpiau, Mira Haegman, Alice Borghi, Laurens Lievens, Srividhya Sundararaman, Amanda Goncalves, Ineke Dhondt, Jorge H PinzonAbstract:The CARD-coiled coil (CC)/Bcl10/MALT1-like Paracaspase (CBM) signaling complexes composed of a CARD-CC family member (CARD-9, -10, -11, or -14), Bcl10, and the type 1 Paracaspase MALT1 (PCASP1) play a pivotal role in immunity, inflammation, and cancer. Targeting MALT1 proteolytic activity is of potential therapeutic interest. However, little is known about the evolutionary origin and the original functions of the CBM complex. Type 1 Paracaspases originated before the last common ancestor of planulozoa (bilaterians and cnidarians). Notably in bilaterians, Ecdysozoa (e.g., nematodes and insects) lacks Bcl10, whereas other lineages have a Bcl10 homolog. A survey of invertebrate CARD-CC homologs revealed such homologs only in species with Bcl10, indicating an ancient common origin of the entire CBM complex. Furthermore, vertebrate-like Syk/Zap70 tyrosine kinase homologs with the ITAM-binding SH2 domain were only found in invertebrate organisms with CARD-CC/Bcl10, indicating that this pathway might be related to the original function of the CBM complex. Moreover, the type 1 Paracaspase sequences from invertebrate organisms that have CARD-CC/Bcl10 are more similar to vertebrate Paracaspases. Functional analysis of protein-protein interactions, NF-kappa B signaling, and CYLD cleavage for selected invertebrate type 1 Paracaspase and Bcl10 homologs supports this scenario and indicates an ancient origin of the CARD-CC/Bcl10/Paracaspase signaling complex. By contrast, many of the known MALT1-associated activities evolved fairly recently, indicating that unknown functions are at the basis of the protein conservation. As a proof-of-concept, we provide initial evidence for a CBM- and NF-kappa B-independent neuronal function of the Caenorhabditis elegans type 1 Paracaspase malt-1. In conclusion, this study shows how evolutionary insights may point at alternative functions of MALT1.
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Ancient Origin of the CARD-Coiled Coil/Bcl10/MALT1-Like Paracaspase Signaling Complex Indicates Unknown Critical Functions.
Frontiers in Immunology, 2018Co-Authors: Jens Staal, Yasmine Driege, Paco Hulpiau, Mira Haegman, Alice Borghi, Laurens Lievens, Srividhya Sundararaman, Amanda Goncalves, Ineke DhondtAbstract:The CARD-coiled coil (CC)/Bcl10/MALT1-like Paracaspase (CBM) signaling complexes composed of a CARD-CC family member (CARD-9, -10, -11, or -14), Bcl10, and the type 1 Paracaspase MALT1 (PCASP1) play a pivotal role in immunity, inflammation, and cancer. Targeting MALT1 proteolytic activity is of potential therapeutic interest. However, little is known about the evolutionary origin and the original functions of the CBM complex. Type 1 Paracaspases originated before the last common ancestor of planulozoa (bilaterians and cnidarians). Notably in bilaterians, Ecdysozoa (e.g., nematodes and insects) lacks Bcl10, whereas other lineages have a Bcl10 homolog. A survey of invertebrate CARD-CC homologs revealed such homologs only in species with Bcl10, indicating an ancient common origin of the entire CBM complex. Furthermore, vertebrate-like Syk/Zap70 tyrosine kinase homologs with the ITAM-binding SH2 domain were only found in invertebrate organisms with CARD-CC/Bcl10, indicating that this pathway might be related to the original function of the CBM complex. Moreover, the type 1 Paracaspase sequences from invertebrate organisms that have CARD-CC/Bcl10 are more similar to vertebrate Paracaspases. Functional analysis of protein-protein interactions, NF-kappa B signaling, and CYLD cleavage for selected invertebrate type 1 Paracaspase and Bcl10 homologs supports this scenario and indicates an ancient origin of the CARD-CC/Bcl10/Paracaspase signaling complex. By contrast, many of the known MALT1-associated activities evolved fairly recently, indicating that unknown functions are at the basis of the protein conservation. As a proof-of-concept, we provide initial evidence for a CBM- and NF-kappa B-independent neuronal function of the Caenorhabditis elegans type 1 Paracaspase malt-1. In conclusion, this study shows how evolutionary insights may point at alternative functions of MALT1.
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GC content of Early Metazoan genes and its impact on gene expression levels in mammalian cell lines
Genome Biology and Evolution, 2018Co-Authors: Jens Staal, Yasmine Driege, Paco Hulpiau, Evi De Keuckelaere, Kai Kamm, Tom Deroo, Ellen Sanders, Katrien Staes, Yvan SaeysAbstract:With the genomes available for many animal clades, including the early-branching metazoans, one can readily study the functional conservation of genes across a diversity of animal lineages. Ectopic expression of an animal protein in, for instance, a mammalian cell line is a generally used strategy in structure–function analysis. However, this might turn out to be problematic in case of distantly related species. Here we analyzed the GC content of the coding sequences of basal animals and show its impact on gene expression levels in human cell lines, and, importantly, how this expression efficiency can be improved. Optimization of the GC3 content in the coding sequences of cadherin, alpha-catenin, and Paracaspase of Trichoplax adhaerens dramatically increased the expression of these basal animal genes in human cell lines.
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the Paracaspase malt1 mediates card14 induced signaling in keratinocytes
EMBO Reports, 2016Co-Authors: Inna S Afonina, Jens Staal, Yasmine Driege, Elien Van Nuffel, Griet Baudelet, Marja Kreike, Rudi BeyaertAbstract:Mutations in CARD14 have recently been linked to psoriasis susceptibility. CARD14 is an epidermal regulator of NF‐κB activation. However, the ability of CARD14 to activate other signaling pathways as well as the biochemical mechanisms that mediate and regulate its function remain to be determined. Here, we report that in addition to NF‐κB signaling, CARD14 activates p38 and JNK MAP kinase pathways, all of which are dependent on the Paracaspase MALT1. Mechanistically, we demonstrate that CARD14 physically interacts with Paracaspase MALT1 and activates MALT1 proteolytic activity and inflammatory gene expression, which are enhanced by psoriasis‐associated CARD14 mutations. Moreover, we show that MALT1 deficiency or pharmacological inhibition of MALT1 catalytic activity inhibits pathogenic mutant CARD14‐induced cytokine and chemokine expression in human primary keratinocytes. Collectively, our findings demonstrate a novel role for MALT1 in CARD14‐induced signaling and indicate MALT1 as a valuable therapeutic target in psoriasis. ![][1] This study shows that the Paracaspase MALT1 is indispensable for CARD14‐induced NF‐κB and MAP kinase signaling. Pharmacological inhibition of MALT1 prevents pro‐inflammatory gene expression in primary keratinocytes induced by psoriasis‐associated mutation of CARD14. EMBO Reports (2016) 17: 914–927 [1]: /embed/graphic-1.gif
Stephan Hailfinger - One of the best experts on this subject based on the ideXlab platform.
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card14 mediated activation of Paracaspase malt1 in keratinocytes implications for psoriasis
Journal of Investigative Dermatology, 2017Co-Authors: Elien Van Nuffel, Rudi Beyaert, Inna S Afonina, Anja Schmitt, Klaus Schulzeosthoff, Stephan HailfingerAbstract:Mutations in caspase recruitment domain-containing protein 14 (CARD14) have been linked to susceptibility to psoriasis. CARD14 is an intracellular scaffold protein that regulates proinflammatory gene expression. Recent studies have offered novel insights into the mechanisms of CARD14-mediated signaling in keratinocytes and the molecular impact of psoriasis-associated CARD14 mutations. CARD14 forms a signaling complex with BCL10 and the Paracaspase MALT1, and this process is enhanced upon pathogenic CARD14 mutation, culminating in the activation of MALT1 protease activity and psoriasis-associated gene expression. This review summarizes the current knowledge of CARD14/MALT1-mediated signaling in keratinocytes and its therapeutic implications in psoriasis.
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targeting b cell lymphomas with inhibitors of the malt1 Paracaspase
Current Opinion in Chemical Biology, 2014Co-Authors: Stephan Hailfinger, Georg Lenz, Margot ThomeAbstract:The Paracaspase MALT1 is an Arg-specific protease that cleaves multiple substrates to promote lymphocyte proliferation and survival. The catalytic activity of MALT1 is normally tightly regulated by antigen receptor triggering, which promotes MALT1 activation by its inducible monoubiquitination-dependent dimerization. Constitutive MALT1 activity is a hallmark of specific subsets of B-cell lymphomas, which are characterized by chromosomal translocations or point mutations that activate MALT1 or its upstream regulators. Recent findings suggest that such lymphomas may be sensitive to treatment with MALT1 inhibitors. Here we review recent progress in the understanding of MALT1 function and regulation, and the development of small molecule MALT1 inhibitors for therapeutic applications.
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detection and measurement of Paracaspase malt1 activity
Methods of Molecular Biology, 2014Co-Authors: Stephan Hailfinger, Christiane Pelzer, Margot ThomeAbstract:The Paracaspase MALT1 is a Cys-dependent, Arg-specific protease that plays an essential role in the activation and proliferation of lymphocytes during the immune response. Oncogenic activation of MALT1 is associated with the development of specific forms of B-cell lymphomas. Through specific cleavage of its substrates, MALT1 controls various aspects of lymphocyte activation, including the activation of transcriptional pathways, the stabilization of mRNAs, and an increase in cellular adhesion. In lymphocytes, the activity of MALT1 is tightly controlled by its inducible monoubiquitination, which promotes the dimerization of MALT1. Here, we describe both in vitro and in vivo assays that have been developed to assess MALT1 activity.
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monoubiquitination and activity of the Paracaspase malt1 requires glutamate 549 in the dimerization interface
PLOS ONE, 2013Co-Authors: Katrin Cabalzar, Christiane Pelzer, Annette Wolf, Georg Lenz, Stephan Hailfinger, Justyna Iwaszkiewicz, Vincent Zoete, Margot ThomeAbstract:The mucosa-associated lymphoid tissue protein-1 (MALT1, also known as Paracaspase) is a protease whose activity is essential for the activation of lymphocytes and the growth of cells derived from human diffuse large B-cell lymphomas of the activated B-cell subtype (ABC DLBCL). Crystallographic approaches have shown that MALT1 can form dimers via its protease domain, but why dimerization is relevant for the biological activity of MALT1 remains largely unknown. Using a molecular modeling approach, we predicted Glu 549 (E549) to be localized within the MALT1 dimer interface and thus potentially relevant. Experimental mutation of this residue into alanine (E549A) led to a complete impairment of MALT1 proteolytic activity. This correlated with an impaired capacity of the mutant to form dimers of the protease domain in vitro, and a reduced capacity to promote NF-κB activation and transcription of the growth-promoting cytokine interleukin-2 in antigen receptor-stimulated lymphocytes. Moreover, this mutant could not rescue the growth of ABC DLBCL cell lines upon MALT1 silencing. Interestingly, the MALT1 mutant E549A was unable to undergo monoubiquitination, which we identified previously as a critical step in MALT1 activation. Collectively, these findings suggest a model in which E549 at the dimerization interface is required for the formation of the enzymatically active, monoubiquitinated form of MALT1.
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Malt1-dependent RelB cleavage promotes canonical NF-κB activation in lymphocytes and lymphoma cell lines
Proceedings of the National Academy of Sciences of the United States of America, 2011Co-Authors: Stephan Hailfinger, Christiane Pelzer, Katrin Cabalzar, Maike Jaworski, Michael Grau, Montserrat Guzzardi, Hendrik Nogai, Jean-enno Charton, Chantal Décaillet, Bernd DörkenAbstract:The protease activity of the Paracaspase Malt1 contributes to antigen receptor-mediated lymphocyte activation and lymphomagenesis. Malt1 activity is required for optimal NF-κB activation, but little is known about the responsible substrate(s). Here we report that Malt1 cleaved the NF-κB family member RelB after Arg-85. RelB cleavage induced its proteasomal degradation and specifically controlled DNA binding of RelA- or c-Rel–containing NF-κB complexes. Overexpression of RelB inhibited expression of canonical NF-κB target genes and led to impaired survival of diffuse large B-cell lymphoma cell lines characterized by constitutive Malt1 activity. These findings identify a central role for Malt1-dependent RelB cleavage in canonical NF-κB activation and thereby provide a rationale for the targeting of Malt1 in immunomodulation and cancer treatment.
