Low-dose decitabine modulates T-cell homeostasis and restores immune tolerance in immune thrombocytopenia
Panpan Han, Yu Hou, Yajing Zhao, Yang Liu, Tianshu Yu, Yunqi Sun, Haoyi Wang, Pengcheng Xu, Guosheng Li, Tao Sun, Xiang Hu, Xinguang Liu, Lizhen Li, Jun Peng, Hai Zhou, Ming Hou, Panpan Han, Yu Hou, Yajing Zhao, Yang Liu, Tianshu Yu, Yunqi Sun, Haoyi Wang, Pengcheng Xu, Guosheng Li, Tao Sun, Xiang Hu, Xinguang Liu, Lizhen Li, Jun Peng, Hai Zhou, Ming Hou
Abstract
Our previous clinical study showed that low-dose decitabine exhibited sustained responses in nearly half of patients with refractory immune thrombocytopenia (ITP). The long-term efficacy of decitabine in ITP is not likely due to its simple role in increasing platelet production. Whether decitabine has the potential to restore immune tolerance in ITP is unknown. In this study, we analyzed the effect of decitabine on T-cell subpopulations in ITP in vitro and in vivo. We found that low-dose decitabine promoted the generation and differentiation of regulatory T (Treg) cells and augmented their immunosuppressive function. Splenocytes from CD61 knockout mice immunized with CD61+ platelets were transferred into severe combined immunodeficient mouse recipients to induce a murine model of ITP. Low-dose decitabine alleviated thrombocytopenia and restored the balance between Treg and helper T (Th) cells in active ITP mice. Treg deletion and depletion offset the effect of decitabine in restoring CD4+ T-cell subpopulations in ITP mice. For patients who received low-dose decitabine, the quantity and function of Treg cells were substantially improved, whereas Th1 and Th17 cells were suppressed compared with the pretreatment levels. Next-generation RNA-sequencing and cytokine analysis showed that low-dose decitabine rebalanced T-cell homeostasis, decreased proinflammatory cytokines, and downregulated phosphorylated STAT3 in patients with ITP. STAT3 inhibition analysis suggested that low-dose decitabine might restore Treg cells by inhibiting STAT3 activation. In conclusion, our data indicate that the immunomodulatory effect of decitabine provides one possible mechanistic explanation for the sustained response achieved by low-dose decitabine in ITP.
© 2021 by The American Society of Hematology.
Figures
References
- Olsson B, Andersson PO, Jernås M, et al. . T-cell-mediated cytotoxicity toward platelets in chronic idiopathic thrombocytopenic purpura. Nat Med. 2003;9(9):1123-1124.
- McMillan R, Wang L, Tomer A, Nichol J, Pistillo J.. Suppression of in vitro megakaryocyte production by antiplatelet autoantibodies from adult patients with chronic ITP. Blood. 2004;103(4):1364-1369.
- Cines DB, Bussel JB, Liebman HA, Luning Prak ET.. The ITP syndrome: pathogenic and clinical diversity. Blood. 2009;113(26):6511-6521.
- Kuter DJ, Rummel M, Boccia R, et al. . Romiplostim or standard of care in patients with immune thrombocytopenia. N Engl J Med. 2010;363(20):1889-1899.
- Imbach P, Crowther M.. Thrombopoietin-receptor agonists for primary immune thrombocytopenia. N Engl J Med. 2011;365(8):734-741.
- Cines DB, Cuker A, Semple JW.. Pathogenesis of immune thrombocytopenia. Presse Med. 2014;43(4 pt 2):e49-e59.
- Nishimoto T, Kuwana M.. CD4+CD25+Foxp3+ regulatory T cells in the pathophysiology of immune thrombocytopenia. Semin Hematol. 2013; 50(suppl 1):S43-S49.
- Son BR, Kim JY.. Association of CD4(+)CD25(+)FoxP3(+) regulatory T cells with natural course of childhood chronic immune thrombocytopenic purpura. Korean J Pediatr. 2015;58(5):178-182.
- Arandi N, Mirshafiey A, Jeddi-Tehrani M, et al. . Alteration in frequency and function of CD4+CD25+FOXP3+ regulatory T cells in patients with immune thrombocytopenic purpura. Iran J Allergy Asthma Immunol. 2014;13(2):85-92.
- Panitsas FP, Theodoropoulou M, Kouraklis A, et al. . Adult chronic idiopathic thrombocytopenic purpura (ITP) is the manifestation of a type-1 polarized immune response. Blood. 2004;103(7):2645-2647.
- McKenzie CGJ, Guo L, Freedman J, Semple JW.. Cellular immune dysfunction in immune thrombocytopenia (ITP). Br J Haematol. 2013;163(1):10-23.
- Li J, Wang Z, Hu S, Zhao X, Cao L.. Correction of abnormal T cell subsets by high-dose dexamethasone in patients with chronic idiopathic thrombocytopenic purpura. Immunol Lett. 2013;154(1-2):42-48.
- Li Z, Mou W, Lu G, et al. . Low-dose rituximab combined with short-term glucocorticoids up-regulates Treg cell levels in patients with immune thrombocytopenia. Int J Hematol. 2011;93(1):91-98.
- Bao W, Bussel JB, Heck S, et al. . Improved regulatory T-cell activity in patients with chronic immune thrombocytopenia treated with thrombopoietic agents. Blood. 2010;116(22):4639-4645.
- Steensma DP.Myelodysplasia, megakaryocytes, and methylation. Leuk Res. 2004;28(8):775-776.
- Wang J, Yi Z, Wang S, Li Z.. The effect of decitabine on megakaryocyte maturation and platelet release. Thromb Haemost. 2011;106(2):337-343.
- Zhou H, Hou Y, Liu X, et al. . Low-dose decitabine promotes megakaryocyte maturation and platelet production in healthy controls and immune thrombocytopenia. Thromb Haemost. 2015;113(5):1021-1034.
- Zhou H, Qin P, Liu Q, et al. . A prospective, multicenter study of low dose decitabine in adult patients with refractory immune thrombocytopenia. Am J Hematol. 2019;94(12):1374-1381.
- Kim HP, Leonard WJ.. CREB/ATF-dependent T cell receptor-induced FoxP3 gene expression: a role for DNA methylation. J Exp Med. 2007;204(7):1543-1551.
- Sánchez-Abarca LI, Gutierrez-Cosio S, Santamaría C, et al. . Immunomodulatory effect of 5-azacytidine (5-azaC): potential role in the transplantation setting. Blood. 2010;115(1):107-121.
- Chow L, Aslam R, Speck ER, et al. . A murine model of severe immune thrombocytopenia is induced by antibody- and CD8+ T cell-mediated responses that are differentially sensitive to therapy. Blood. 2010;115(6):1247-1253.
- Hou Y, Feng Q, Xu M, et al. . High-dose dexamethasone corrects impaired myeloid-derived suppressor cell function via Ets1 in immune thrombocytopenia. Blood. 2016;127(12):1587-1597.
- Aslam R, Hu Y, Gebremeskel S, et al. . Thymic retention of CD4+CD25+FoxP3+ T regulatory cells is associated with their peripheral deficiency and thrombocytopenia in a murine model of immune thrombocytopenia. Blood. 2012;120(10):2127-2132.
- Liu W, Li H, Hao Y, et al. . Decreased immunosuppressive actions of 1α, 25-dihydroxyvitamin D3 in patients with immune thrombocytopenia. Mol Immunol. 2016;78:89-97.
- Chen M, Yan R, Zhou K, et al. . Akt-mediated platelet apoptosis and its therapeutic implications in immune thrombocytopenia. Proc Natl Acad Sci U S A. 2018;115(45):E10682-E10691.
- Hu Y, Wang X, Yu S, Hou Y, Ma D, Hou M.. Neutralizations of IL-17A and IL-21 regulate regulatory T cell/T-helper 17 imbalance via T-helper 17-associated signaling pathway in immune thrombocytopenia. Expert Opin Ther Targets. 2015;19(6):723-732.
- Cuker A, Neunert CE.. How I treat refractory immune thrombocytopenia. Blood. 2016;128(12):1547-1554.
- Lambert MP, Gernsheimer TB.. Clinical updates in adult immune thrombocytopenia. Blood. 2017;129(21):2829-2835.
- Li H, Xuan M, Yang R.. DNA methylation and primary immune thrombocytopenia. Semin Hematol. 2013;50(suppl 1):S116-S126.
- Chen Z, Guo Z, Ma J, Ma J, Liu F, Wu R.. Foxp3 methylation status in children with primary immune thrombocytopenia. Hum Immunol. 2014;75(11):1115-1119.
- Chen ZP, Gu DS, Zhou ZP, et al. . Decreased expression of MBD2 and MBD4 gene and genomic-wide hypomethylation in patients with primary immune thrombocytopenia. Hum Immunol. 2011;72(6):486-491.
- Han P, Yu T, Hou Y, et al. . Low-dose decitabine inhibits cytotoxic T lymphocytes-mediated platelet destruction via modulating PD-1 methylation in immune thrombocytopenia. Front Immunol. 2021;12:630693.
- Wang L, Amoozgar Z, Huang J, et al. . Decitabine enhances lymphocyte migration and function and synergizes with CTLA-4 blockade in a murine ovarian cancer model. Cancer Immunol Res. 2015;3(9):1030-1041.
- Choi J, Ritchey J, Prior JL, et al. . In vivo administration of hypomethylating agents mitigate graft-versus-host disease without sacrificing graft-versus-leukemia. Blood. 2010;116(1):129-139.
- Mangano K, Fagone P, Bendtzen K, et al. . Hypomethylating agent 5-aza-2'-deoxycytidine (DAC) ameliorates multiple sclerosis in mouse models. J Cell Physiol. 2014;229(12):1918-1925.
- Wang X, Wang J, Yu Y, et al. . Decitabine inhibits T cell proliferation via a novel TET2-dependent mechanism and exerts potent protective effect in mouse auto- and allo-immunity models. Oncotarget. 2017;8(34):56802-56815.
- Zheng Q, Xu Y, Liu Y, et al. . Induction of Foxp3 demethylation increases regulatory CD4+CD25+ T cells and prevents the occurrence of diabetes in mice. J Mol Med (Berl). 2009;87(12):1191-1205.
- Fagone P, Mazzon E, Chikovani T, et al. . Decitabine induces regulatory T cells, inhibits the production of IFN-gamma and IL-17 and exerts preventive and therapeutic efficacy in rodent experimental autoimmune neuritis. J Neuroimmunol. 2018;321:41-48.
- Guo L, Kapur R, Aslam R, et al. . CD20+ B-cell depletion therapy suppresses murine CD8+ T-cell-mediated immune thrombocytopenia. Blood. 2016;127(6):735-738.
- Stasi R, Cooper N, Del Poeta G, et al. . Analysis of regulatory T-cell changes in patients with idiopathic thrombocytopenic purpura receiving B cell-depleting therapy with rituximab. Blood. 2008;112(4):1147-1150.
- Ling Y, Cao X, Yu Z, Ruan C.. Circulating dendritic cells subsets and CD4+Foxp3+ regulatory T cells in adult patients with chronic ITP before and after treatment with high-dose dexamethasome. Eur J Haematol. 2007;79(4):310-316.
- Wang SC, Yang KD, Lin CY, et al. . Intravenous immunoglobulin therapy enhances suppressive regulatory T cells and decreases innate lymphoid cells in children with immune thrombocytopenia. Pediatr Blood Cancer. 2020;67(2):e28075.
- Guo L, Yang L, Speck ER, et al. . Allogeneic platelet transfusions prevent murine T-cell-mediated immune thrombocytopenia. Blood. 2014;123(3):422-427.
- Semple JW, Milev Y, Cosgrave D, et al. . Differences in serum cytokine levels in acute and chronic autoimmune thrombocytopenic purpura: relationship to platelet phenotype and antiplatelet T-cell reactivity. Blood. 1996;87(10):4245-4254.
- Qu MM, Liu XN, Liu XG, et al. . Cytokine changes in response to TPO receptor agonist treatment in primary immune thrombocytopenia. Cytokine. 2017;92:110-117.
- Andersson PO, Olsson A, Wadenvik H.. Reduced transforming growth factor-beta1 production by mononuclear cells from patients with active chronic idiopathic thrombocytopenic purpura. Br J Haematol. 2002;116(4):862-867.
- Guo C, Chu X, Shi Y, et al. . Correction of Th1-dominant cytokine profiles by high-dose dexamethasone in patients with chronic idiopathic thrombocytopenic purpura. J Clin Immunol. 2007;27(6):557-562.
- Villarino AV, Kanno Y, O’Shea JJ.. Mechanisms and consequences of Jak-STAT signaling in the immune system. Nat Immunol. 2017;18(4):374-384.
- Laurence A, Amarnath S, Mariotti J, et al. . STAT3 transcription factor promotes instability of nTreg cells and limits generation of iTreg cells during acute murine graft-versus-host disease. Immunity. 2012;37(2):209-222.
- Burchill MA, Yang J, Vogtenhuber C, Blazar BR, Farrar MA.. IL-2 receptor beta-dependent STAT5 activation is required for the development of Foxp3+ regulatory T cells. J Immunol. 2007;178(1):280-290.
- Yao Z, Kanno Y, Kerenyi M, et al. . Nonredundant roles for Stat5a/b in directly regulating Foxp3. Blood. 2007;109(10):4368-4375.
- Zorn E, Nelson EA, Mohseni M, et al. . IL-2 regulates FOXP3 expression in human CD4+CD25+ regulatory T cells through a STAT-dependent mechanism and induces the expansion of these cells in vivo. Blood. 2006;108(5):1571-1579.
- Fujino M, Li XK.. Role of STAT3 in regulatory T lymphocyte plasticity during acute graft-vs.-host-disease. JAK-STAT. 2013;2(4):e24529.
- Sailer T, Lechner K, Panzer S, Kyrle PA, Pabinger I.. The course of severe autoimmune thrombocytopenia in patients not undergoing splenectomy. Haematologica. 2006;91(8):1041-1045.
- Schifferli A, Holbro A, Chitlur M, et al. ; Intercontinental Cooperative ITP Study Group (ICIS). A comparative prospective observational study of children and adults with immune thrombocytopenia: 2-year follow-up. Am J Hematol. 2018;93(6):751-759.
- Neunert C, Terrell DR, Arnold DM, et al. . American Society of Hematology 2019 guidelines for immune thrombocytopenia [published correction appears in Blood Adv. 2020;4(2):252]. Blood Adv. 2019;3(23):3829-3866.
- Provan D, Arnold DM, Bussel JB, et al. . Updated international consensus report on the investigation and management of primary immune thrombocytopenia. Blood Adv. 2019;3(22):3780-3817.
Source: PubMed