CAMK2G is identified as a novel therapeutic target for myelofibrosis

Blood Advances - Tập 6 - Trang 1585-1597 - 2022
Masashi Miyauchi1, Ken Sasaki1, Yuki Kagoya1, Kazuki Taoka1, Yosuke Masamoto1, Sho Yamazaki1, Shunya Arai1, Hideaki Mizuno1, Mineo Kurokawa1
1Department of Hematology and Oncology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

Tóm tắt

Abstract Although JAK1/2 inhibition is effective in alleviating symptoms of myelofibrosis (MF), it does not result in the eradication of MF clones, which can lead to inhibitor-resistant clones emerging during the treatment. Here, we established induced pluripotent stem cells (iPSCs) derived from MF patient samples (MF-iPSCs) harboring JAK2 V617F, CALR type 1, or CALR type 2 mutations. We demonstrated that these cells faithfully recapitulate the drug sensitivity of the disease. These cells were used for chemical screening, and calcium/calmodulin-dependent protein kinase 2 (CAMK2) was identified as a promising therapeutic target. MF model cells and mice induced by MPL W515L, another type of mutation recurrently detected in MF patients, were used to elucidate the therapeutic potential of CAMK2 inhibition. CAMK2 inhibition was effective against JAK2 inhibitor-sensitive and JAK2 inhibitor-resistant cells. Further research revealed CAMK2 γ subtype was important in MF model cells induced by MPL W515L. We showed that CAMK2G hetero knockout in the primary bone marrow cells expressing MPL W515L decreased colony-forming capacity. CAMK2G inhibition with berbamine, a CAMK2G inhibitor, significantly prolonged survival and reduced disease phenotypes, such as splenomegaly and leukocytosis in a MF mouse model induced by MPL W515L. We investigated the molecular mechanisms underlying the therapeutic effect of CAMK2G inhibition and found that CAMK2G is activated by MPL signaling in MF model cells and is an effector in the MPL-JAK2 signaling pathway in these cells. These results indicate CAMK2G plays an important role in MF, and CAMK2G inhibition may be a novel therapeutic strategy that overcomes resistance to JAK1/2 inhibition.

Tài liệu tham khảo

Vainchenker, 2017, Genetic basis and molecular pathophysiology of classical myeloproliferative neoplasms, Blood., 129, 667, 10.1182/blood-2016-10-695940 Cervantes, 2009, New prognostic scoring system for primary myelofibrosis based on a study of the International Working Group for Myelofibrosis Research and Treatment, Blood., 113, 2895, 10.1182/blood-2008-07-170449 Zahr, 2016, Bone marrow fibrosis in myelofibrosis: pathogenesis, prognosis and targeted strategies, Haematologica., 101, 660, 10.3324/haematol.2015.141283 Kroeger, 2007, Dose-reduced conditioning followed by allogeneic stem cell transplantation in patients with myelofibrosis. results from a multicenter prospective trial of the Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation (EBMT) [abstract], Blood., 110 Harrison, 2012, JAK inhibition with ruxolitinib versus best available therapy for myelofibrosis, N Engl J Med., 366, 787, 10.1056/NEJMoa1110556 Verstovsek, 2012, A double-blind, placebo-controlled trial of ruxolitinib for myelofibrosis, N Engl J Med., 366, 799, 10.1056/NEJMoa1110557 Verstovsek, 2017, Long-term survival in patients treated with ruxolitinib for myelofibrosis: COMFORT-I and -II pooled analyses, J Hematol Oncol., 10, 156, 10.1186/s13045-017-0527-7 Pardanani, 2015, Safety and efficacy of fedratinib in patients with primary or secondary myelofibrosis: a randomized clinical trial, JAMA Oncol., 1, 643, 10.1001/jamaoncol.2015.1590 Pardanani, 2011, Safety and efficacy of TG101348, a selective JAK2 inhibitor, in myelofibrosis, J Clin Oncol., 29, 789, 10.1200/JCO.2010.32.8021 Gotlib, 2021, Long-term follow up of a phase 1/2 study of SAR302503, an oral JAK2 selective inhibitor, in patients with myelofibrosis (MF) Koppikar, 2012, Heterodimeric JAK-STAT activation as a mechanism of persistence to JAK2 inhibitor therapy, Nature., 489, 155, 10.1038/nature11303 Araki, 2016, Activation of the thrombopoietin receptor by mutant calreticulin in CALR-mutant myeloproliferative neoplasms, Blood., 127, 1307, 10.1182/blood-2015-09-671172 Takayama, 2008, Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors, Blood., 111, 5298, 10.1182/blood-2007-10-117622 Ye, 2009, Human-induced pluripotent stem cells from blood cells of healthy donors and patients with acquired blood disorders, Blood., 114, 5473, 10.1182/blood-2009-04-217406 Saliba, 2013, Heterozygous and homozygous JAK2(V617F) states modeled by induced pluripotent stem cells from myeloproliferative neoplasm patients, PLoS One., 8, e74257, 10.1371/journal.pone.0074257 Kumano, 2012, Generation of induced pluripotent stem cells from primary chronic myelogenous leukemia patient samples, Blood., 119, 6234, 10.1182/blood-2011-07-367441 Hosoi, 2014, Generation of induced pluripotent stem cells derived from primary and secondary myelofibrosis patient samples, Exp Hematol., 42, 816, 10.1016/j.exphem.2014.03.010 Backs, 2010, The γ isoform of CaM kinase II controls mouse egg activation by regulating cell cycle resumption, Proc Natl Acad Sci USA., 107, 81, 10.1073/pnas.0912658106 Okita, 2013, An efficient nonviral method to generate integration-free human-induced pluripotent stem cells from cord blood and peripheral blood cells, Stem Cells., 31, 458, 10.1002/stem.1293 Okita, 2011, A more efficient method to generate integration-free human iPS cells, Nat Methods., 8, 409, 10.1038/nmeth.1591 Takayama, 2010, Transient activation of c-MYC expression is critical for efficient platelet generation from human induced pluripotent stem cells, J Exp Med., 207, 2817, 10.1084/jem.20100844 Sato, 2014, Evi1 defines leukemia-initiating capacity and tyrosine kinase inhibitor resistance in chronic myeloid leukemia, Oncogene., 33, 5028, 10.1038/onc.2014.108 Rampal, 2014, Integrated genomic analysis illustrates the central role of JAK-STAT pathway activation in myeloproliferative neoplasm pathogenesis, Blood., 123, e123, 10.1182/blood-2014-02-554634 Fiskus, 2011, Heat shock protein 90 inhibitor is synergistic with JAK2 inhibitor and overcomes resistance to JAK2-TKI in human myeloproliferative neoplasm cells, Clin Cancer Res., 17, 7347, 10.1158/1078-0432.CCR-11-1541 Gu, 2017, Stabilization of the c-Myc protein by CAMKIIγ promotes T cell lymphoma, Cancer Cell., 32, 115, 10.1016/j.ccell.2017.06.001 Gu, 2012, CaMKII γ, a critical regulator of CML stem/progenitor cells, is a target of the natural product berbamine, Blood., 120, 4829, 10.1182/blood-2012-06-434894 Pikman, 2006, MPLW515L is a novel somatic activating mutation in myelofibrosis with myeloid metaplasia, PLoS Med., 3, e270, 10.1371/journal.pmed.0030270 Ma, 2014, γCaMKII shuttles Ca2+/CaM to the nucleus to trigger CREB phosphorylation and gene expression, Cell., 159, 281, 10.1016/j.cell.2014.09.019 Park, 2008, Disease-specific induced pluripotent stem cells, Cell., 134, 877, 10.1016/j.cell.2008.07.041 Malik, 2014, Mission CaMKIIγ: shuttle calmodulin from membrane to nucleus, Cell., 159, 235, 10.1016/j.cell.2014.09.023 Gu, 2016, Aberrant activation of CaMKIIγ accelerates chronic myeloid leukemia blast crisis, Leukemia., 30, 1282, 10.1038/leu.2016.53 Si, 2008, Activated Ca2+/calmodulin-dependent protein kinase IIgamma is a critical regulator of myeloid leukemia cell proliferation, Cancer Res., 68, 3733, 10.1158/0008-5472.CAN-07-2509 Chi, 2016, Phosphorylation of calcium/calmodulin-stimulated protein kinase II at T286 enhances invasion and migration of human breast cancer cells, Sci Rep., 6, 33132, 10.1038/srep33132 Chai, 2015, Ca2+/calmodulin-dependent protein kinase IIγ enhances stem-like traits and tumorigenicity of lung cancer cells, Oncotarget., 6, 16069, 10.18632/oncotarget.3866 Seow, 1992, Comparative effects of tetrandrine and berbamine on production of the inflammatory cytokines interleukin-1 and tumor necrosis factor, Life Sci., 50, PL53, 10.1016/0024-3205(92)90373-W Ren, 2008, Novel immunomodulatory properties of berbamine through selective down-regulation of STAT4 and action of IFN-γ in experimental autoimmune encephalomyelitis, J Immunol., 181, 1491, 10.4049/jimmunol.181.2.1491 Xu, 2006, Berbamine: a novel inhibitor of bcr/abl fusion gene with potent anti-leukemia activity, Leuk Res., 30, 17, 10.1016/j.leukres.2005.05.023 Xie, 2009, Berbamine derivatives: a novel class of compounds for anti-leukemia activity, Eur J Med Chem., 44, 3293, 10.1016/j.ejmech.2009.02.018 Dong, 2004, Study on effect of berbamine on multidrug resistance leukemia K562/Adr cells [in Chinese], Zhongguo Zhong Xi Yi Jie He Za Zhi., 24, 820 Grisouard, 2014, Selective deletion of Jak2 in adult mouse hematopoietic cells leads to lethal anemia and thrombocytopenia, Haematologica., 99, e52, 10.3324/haematol.2013.100016 Wang, 2015, The emerging role of CaMKII in cancer, Oncotarget., 6, 11725, 10.18632/oncotarget.3955