SET domain containing 2 promotes megakaryocyte polyploidization and platelet generation through methylation of α-tubulin

Lei Chen; Jingkun Liu; Kunying Chen; Yanxun Su; Yihe Chen; Ying Lei; Jia Si; Jie Zhang; Zhaojun Zhang; Weiguo Zou; Xiaohui Zhang; Matthew T Rondina; Qian-Fei Wang; Yueying Li

doi:10.1016/j.jtha.2024.03.010

SET domain containing 2 promotes megakaryocyte polyploidization and platelet generation through methylation of α-tubulin

J Thromb Haemost. 2024 Mar 25:S1538-7836(24)00168-5. doi: 10.1016/j.jtha.2024.03.010. Online ahead of print.

Authors

Lei Chen¹, Jingkun Liu¹, Kunying Chen¹, Yanxun Su¹, Yihe Chen¹, Ying Lei¹, Jia Si¹, Jie Zhang¹, Zhaojun Zhang², Weiguo Zou³, Xiaohui Zhang⁴, Matthew T Rondina⁵, Qian-Fei Wang⁶, Yueying Li⁷

Affiliations

¹ Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences and China National Center for Bioinformation, Beijing, China; University of Chinese Academy of Sciences, Beijing, China.
² University of Chinese Academy of Sciences, Beijing, China; Key Laboratory of Genome Science and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, China National Center of Bioinformation, Beijing, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China.
³ Shanghai Institute of Microsurgery on Extremities, and Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China; State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Sciences, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, China.
⁴ Peking University People's Hospital, Peking University Institute of Hematology, Beijing, China; National Clinical Research Center for Hematologic Disease, Beijing, China; Beijing Key Laboratory of Hematopoietic Stem Cell Transplantation, Beijing, China; Collaborative Innovation Center of Hematology, Peking University, Beijing, China.
⁵ Departments of Internal Medicine and Pathology, Molecular Medicine Program, University of Utah, Salt Lake City, Utah, USA; Department of Internal Medicine and the Geriatric Research, Education, and Clinical Center, George E. Wahlen Veterans Affairs Medical Center, Salt Lake City, Utah, USA. Electronic address: matthew.rondina@hsc.utah.edu.
⁶ Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences and China National Center for Bioinformation, Beijing, China; University of Chinese Academy of Sciences, Beijing, China. Electronic address: liyy@big.ac.cn.
⁷ Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences and China National Center for Bioinformation, Beijing, China; University of Chinese Academy of Sciences, Beijing, China. Electronic address: wangqf@big.ac.cn.

PMID: 38537781
DOI: 10.1016/j.jtha.2024.03.010

Abstract

Background: Megakaryocytes (MKs) are polyploid cells responsible for producing ∼10¹¹ platelets daily in humans. Unraveling the mechanisms regulating megakaryopoiesis holds the promise for the production of clinical-grade platelets from stem cells, overcoming significant current limitations in platelet transfusion medicine. Previous work identified that loss of the epigenetic regulator SET domain containing 2 (SETD2) was associated with an increased platelet count in mice. However, the role of SETD2 in megakaryopoiesis remains unknown.

Objectives: Here, we examined how SETD2 regulated MK development and platelet production using complementary murine and human systems.

Methods: We manipulated the expression of SETD2 in multiple in vitro and ex vivo models to assess the ploidy of MKs and the function of platelets.

Results: The genetic ablation of Setd2 increased the number of high-ploidy bone marrow MKs. Peripheral platelet counts in Setd2 knockout mice were significantly increased ∼2-fold, and platelets exhibited normal size, morphology, and function. By knocking down and overexpressing SETD2 in ex vivo human cell systems, we demonstrated that SETD2 negatively regulated MK polyploidization by controlling methylation of α-tubulin, microtubule polymerization, and MK nuclear division. Small-molecule inactivation of SETD2 significantly increased the production of high-ploidy MKs and platelets from human-induced pluripotent stem cells and cord blood CD34⁺ cells.

Conclusion: These findings identify a previously unrecognized role for SETD2 in regulating megakaryopoiesis and highlight the potential of targeting SETD2 to increase platelet production from human cells for transfusion practices.

Keywords: SETD2; megakaryocyte; platelet; polyploidy; α-tubulin.