1. BTK suppresses myeloma cellular senescence through activating AKT/P27/Rb signaling
Ye Yang, Yue Lu, Wen Zhang, Hongbao Yang, Gang Yin, Hailin Peng, Zhidan Tian, Sicheng Lu, Yi Zhang, Chunyan Gu Oncotarget . 2017 May 23;8(34):56858-56867. doi: 10.18632/oncotarget.18096.
We previously explored the role of BTK in maintaining multiple myeloma stem cells (MMSCs) self-renewal and drug-resistance. Here we investigated the elevation of BTK suppressing MM cellular senescence, a state of irreversible cellular growth arrest. We firstly discovered that an increased expression of BTK in MM samples compared to normal controls by immunohistochemistry (IHC), and significant chromosomal gain in primary samples. In addition, BTK high-expressing MM patients are associated with poor outcome in both Total Therapy 2 (TT2) and TT3 cohorts. Knockdown BTK expression by shRNA induced MM cellular senescence using β-galactosidase (SA-b-gal) staining, cell growth arrest by cell cycle staining and decreased clonogenicity while forcing BTK expression in MM cells abrogated these characteristics. We also validated this feature in mouse embryonic fibroblast cells (MEFs), which showed that elevated BTK expression was resistant to MEF senescence after serial cultivationin vitro. Further mechanism study revealed that BTK activated AKT signaling leading to down-regulation of P27 expression and hindered RB activity while AKT inhibitor, LY294002, overcame BTK-overexpression induced cellular senescence resistance. Eventually we demonstrated that BTK inhibitor, CGI-1746, induced MM cellular senescence, colony reduction and tumorigenecity inhibitionin vivo. Summarily, we designate a novel mechanism of BTK in mediating MM growth, and BTK inhibitor is of great potentialin vivoandin vitrosuggesting BTK is a promising therapeutic target for MM.
2. Bruton tyrosine kinase is a therapeutic target in stem-like cells from multiple myeloma
Dana Levasseur, Ye Yang, Junwei Huang, Erik Wendlandt, Yi Tao, Mu Hao, Fenghuang Zhan, Siegfried Janz, Mohamed E Salama, Reinaldo Franqui, Zhimin Gu, Satyabrata Das, Guido Tricot, Hongwei Xu, Jumei Shi Cancer Res . 2015 Feb 1;75(3):594-604. doi: 10.1158/0008-5472.CAN-14-2362.
Ibrutinib (Imbruvica), a small-drug inhibitor of Bruton tyrosine kinase (BTK), is currently undergoing clinical testing in patients with multiple myeloma, yet important questions on the role of BTK in myeloma biology and treatment are outstanding. Using flow-sorted side population cells from human myeloma cell lines and multiple myeloma primary samples as surrogate for the elusive multiple myeloma stem cell, we found that elevated expression of BTK in myeloma cells leads to AKT/WNT/β-catenin-dependent upregulation of key stemness genes (OCT4, SOX2, NANOG, and MYC) and enhanced self-renewal. Enforced transgenic expression of BTK in myeloma cells increased features of cancer stemness, including clonogenicity and resistance to widely used myeloma drugs, whereas inducible knockdown of BTK abolished them. Furthermore, overexpression of BTK in myeloma cells promoted tumor growth in laboratory mice and rendered side population-derived tumors that contained high levels of BTK more sensitive to the selective, second-generation BTK inhibitor, CGI1746, than side population-derived tumors that harbored low levels of BTK. Taken together, these findings implicate BTK as a positive regulator of myeloma stemness and provide additional support for the clinical testing of BTK-targeted therapies in patients with myeloma.
3. Specific Btk inhibition suppresses B cell- and myeloid cell-mediated arthritis
Steven L Gallion, James Barbosa, Wyne P Lee, Patricia M Maciejewski, Wendy B Young, Douglas R Davies, Sherry Yeh, Bart L Staker, Peter Gribling, J Andrew Whitney, Anthony M Giannetti, Christine Yu, Vincent Hurez, Mercedesz Balazs, Karin Reif, Laura E DeForge, Randall Jones, Jeffrey E Kropf, Sarah G Hymowitz, Brandon J Bravo, Julie A Di Paolo, Lauri Diehl, James Darrow, Juan Zhang, Tao Huang, Ronald Ferrando, Richard A D Carano, Scott A Mitchell, Kevin S Currie, Kai H Barck, Hong Rong Nat Chem Biol . 2011 Jan;7(1):41-50. doi: 10.1038/nchembio.481.
Bruton's tyrosine kinase (Btk) is a therapeutic target for rheumatoid arthritis, but the cellular and molecular mechanisms by which Btk mediates inflammation are poorly understood. Here we describe the discovery of CGI1746, a small-molecule Btk inhibitor chemotype with a new binding mode that stabilizes an inactive nonphosphorylated enzyme conformation. CGI1746 has exquisite selectivity for Btk and inhibits both auto- and transphosphorylation steps necessary for enzyme activation. Using CGI1746, we demonstrate that Btk regulates inflammatory arthritis by two distinct mechanisms. CGI1746 blocks B cell receptor-dependent B cell proliferation and in prophylactic regimens reduces autoantibody levels in collagen-induced arthritis. In macrophages, Btk inhibition abolishes FcγRIII-induced TNFα, IL-1β and IL-6 production. Accordingly, in myeloid- and FcγR-dependent autoantibody-induced arthritis, CGI1746 decreases cytokine levels within joints and ameliorates disease. These results provide new understanding of the function of Btk in both B cell- or myeloid cell-driven disease processes and provide a compelling rationale for targeting Btk in rheumatoid arthritis.
