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MYELODYSPLASTIC NEOPLASM

Aberrant splicing of CHEK1 is a driver of megakaryocytic dysplasia in U2AF1S34F mutant myelodysplastic neoplasms

Leukemia volume 39, pages 2246–2255 (2025)Cite this article

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Abstract

U2AF1 mutations are common in patients with myelodysplastic neoplasms (MDS), suggesting that aberrant splicing of pre-mRNAs driven by mutant U2AF1 could play a critical role in MDS pathogenesis. Previous studies have demonstrated that U2AF1S34F mutation impairs the differentiation of erythrocytes and granulocytes, but the impact on megakaryocytes (MKs) remains unclear. Here, by integrating data from MDS patients and cell lines with U2AF1 mutations, we determined that U2AF1 mutations are associated with dysmegakaryopoiesis, induce the generation of abnormal MKs, especially micro-MKs, and induce significant thrombocytopenia. We determined that mutant U2AF1-mediated aberrant splicing of DNA biosynthesis-related genes, such as CHEK1, is required for normal MK polyploidization. The mis-splicing of CHEK1, in turn, accounts for the increased number of abnormal MKs in U2AF1-mutant MDS patients. Moreover, U2AF1S34 mutations induce the deficiency of CHK1 and the activation of its phosphorylation, thereby further driving the impairment of MK polyploidization and maturation. Accordingly, treatment with selective CHK1 inhibitor significantly reduces abnormal MK production in vitro. Taken together, these findings demonstrate that U2AF1 mutations induce the generation of abnormal MKs by driving aberrant splicing of the CHEK1 cell cycle-related gene, revealing the molecular basis for dysmegakaryopoiesis in MDS and identifying a new potential target for MDS treatment.

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Fig. 1: U2AF1S34 mutations are significantly associated with the generation of micro-megakayrocytes (micro-MKs) in MDS patients.
Fig. 2: The U2AF1S34F mutation impairs the polyploidization of the K562 and HEL cell lines.
Fig. 3: U2AF1 mutation results in abnormal splicing of DNA biosynthesis-related genes.
Fig. 4: Functional effects of CHEK1 knockdown or CHEK1-S expression on MK polyploidization.
Fig. 5: CHK1 inhibition rescues the defects in MK polyploidization observed in MDS U2AF1S34.

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Data availability

The datasets supporting the findings of this study are available from the corresponding authors upon reasonable request.

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Acknowledgements

We would like to express our sincere appreciation to Professors Omar Abdel-Wahab and Kevin Rouault-Pierre for their invaluable guidance and support throughout this research. Their expertise and constructive feedback have been instrumental in the development of this study.

Funding

This study is supported in part by National Natural Science Funds (Nos. 82170139, 81530008 and 82070134), CAMS Initiative Fund for Medical Sciences (Nos. 2023-I2M-2-007, 2022-I2M-1-022,), Clinical research fund from National Clinical Research Centre for Blood Diseases (Nos. 2023NCRCA0117 and 2023NCRCA0103), Medical and Healthy Science Innovative Program of Chinese Academy of Medical Science (2022-I2M-C&T-B-093).

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Author notes

  1. These authors contributed equally: Wenjun Zhang, Bing Li, Jinqin Liu, Yiru Yan

  2. These authors jointly supervised this work: Bing Li, Zhijian Xiao

Authors and Affiliations

  1. State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China

    Wenjun Zhang, Bing Li, Jinqin Liu, Yiru Yan, Lin Yang, Zefeng Xu, Hongtao Wang, Lihong Shi, Jiaxi Zhou & Zhijian Xiao

  2. MDS and MPN Centre, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China

    Wenjun Zhang, Bing Li, Yiru Yan, Tiejun Qin, Zefeng Xu & Zhijian Xiao

  3. Hematologic Pathology Center, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China

    Qi Sun, Yujiao Jia, Huijun Wang & Zhijian Xiao

  4. Department of Cell Systems and Anatomy, Department of Pathology and Laboratory Medicine, UT Health San Antonio, Joe R. and Teresa Lozano Long School of Medicine, Mays Cancer Center at UT Health San Antonio, San Antonio, TX, USA

    Gang Huang

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Contributions

ZJX and BL initiated the concept for the study. ZJX and BL designed the study. WJZ performed the research, collected data, write and revised manuscript, JQL, BL, LY and YRY performed the research. TJQ, ZFX, QS, YJJ and HJW recruited patients and collected data. GH, LHS, HTW and JXZ provided support for the experiments. WJZ, BL and ZJX prepared the typescript. All authors read, approved the final manuscript, and agreed to submit for publication.

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Correspondence to Bing Li or Zhijian Xiao.

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Zhang, W., Li, B., Liu, J. et al. Aberrant splicing of CHEK1 is a driver of megakaryocytic dysplasia in U2AF1S34F mutant myelodysplastic neoplasms. Leukemia 39, 2246–2255 (2025). https://doi.org/10.1038/s41375-025-02684-6

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