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Thromb Haemost 2021; 121(02): 192-205
DOI: 10.1055/s-0040-1716530
Cellular Haemostasis and Platelets

Ibrutinib Suppresses Early Megakaryopoiesis but Enhances Proplatelet Formation

Jiansong Huang  ‡   *
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Shujuan Huang‡   *
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Zhixin Ma
4   Clinical Prenatal Diagnosis Center, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Xiangjie Lin
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Xia Li
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Xin Huang
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Jinghan Wang
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Wenle Ye
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Yang Li
5   Department of Obstetrics, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Daqiang He
6   Department of Laboratory Medicine, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Min Yang
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Jiajia Pan
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Qing Ling
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Fenglin Li
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Shihui Mao
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Huafeng Wang
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Yungui Wang
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
,
Jie Jin 
1   Department of Hematology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
2   Key Laboratory of Hematologic Malignancies, Diagnosis and Treatment, Hangzhou, Zhejiang Province, China
3   Institute of Hematology, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
› Author Affiliations Funding This study was supported by grants from the National Natural Science Foundation of China (81820108004 and 81800146) and the Natural Science Foundation of Zhejiang Province (LY20H080008, LY19H080009, and LY16H160009).
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Abstract

Ibrutinib, an irreversible inhibitor of Bruton's tyrosine kinase, has a favorable safety profile in patients with B cell-related malignancies. A primary adverse effect of ibrutinib is thrombocytopenia in the early stages of treatment, but platelet counts increase or recover as treatment continues. Currently, the effects of ibrutinib on megakaryopoiesis remain unclear. In this study, we investigated the mechanism by which ibrutinib induces thrombocytopenia using cord blood CD34+ hematopoietic stem cells (HSCs), a human megakaryoblastic cell line (SET-2), and C57BL/6 mice. We show that treatment with ibrutinib can suppress CD34+ HSC differentiation into megakaryocytes (MKs) and decrease the number of colony-forming unit-MKs (CFU-MKs). The ibrutinib-dependent inhibition of early megakaryopoiesis seems to mainly involve impaired proliferation of progenitor cells without induction of apoptosis. The effects of ibrutinib on late-stage megakaryopoiesis, in contrast to early-stage megakaryopoiesis, include enhanced MK differentiation, ploidy, and proplatelet formation in CD34+ HSC-derived MKs and SET-2 cells. We also demonstrated that MK adhesion and spreading, but not migration, were inhibited by ibrutinib. Furthermore, we revealed that integrin αIIbβ3 outside-in signaling in MKs was inhibited by ibrutinib. Consistent with previous clinical observations, in C57BL/6 mice treated with ibrutinib, platelet counts decreased by days 2 to 7 and recovered to normal levels by day 15. Together, these results reveal the pathogenesis of ibrutinib-induced transient thrombocytopenia. In conclusion, ibrutinib suppresses early megakaryopoiesis, as evidenced by inhibition of MK progenitor cell proliferation and CFU-MK formation. Ibrutinib enhances MK differentiation, ploidy, and proplatelet formation, while it impairs integrin αIIbβ3 outside-in signaling.

Keywords

ibrutinib - megakaryopoiesis - proplatelet formation - thrombocytopenia

Authors' Contributions

J.H. conceived and designed the study, performed the experiments, analyzed the data, and wrote the manuscript; S.H., Z.M., X.L., X.L, X.H., W.Y., D.H., J.P., Q.L., F.L., and S.M. performed the experiments, analyzed the data, and participated in the manuscript preparation; J.W., Y.L., M.Y., Y.W., and H.W. contributed to the data analysis; and J.J. supervised the study, contributed to the design of experiments, analyzed the data, and revised the manuscript.


* These authors contributed equally to this work.


Supplementary Material



Publication History

Received: 26 November 2020

Accepted: 28 July 2020

Article published online:
22 September 2020

© 2020. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

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