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Atp8a1 deletion increases the proliferative activity of hematopoietic stem cells by impairing PTEN function

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Abstract

Purpose

The eukaryotic cell plasma membrane contains several asymmetrically distributed phospholipids, which is maintained by the P4-ATPase flippase complex. Herein, we demonstrated the biological effects and mechanisms of asymmetrical loss in hematopoietic stem cells (HSCs).

Methods

An Atp8a1 knockout mouse model was employed, from which the HSC (long-term HSCs and short-term HSCs) population was analyzed to assess their abundance and function. Additionally, competitive bone marrow transplantation and 5-FU stress assays were performed. RNA sequencing was performed on Hematopoietic Stem and Progenitor Cells, and DNA damage was assayed using immunofluorescence staining and comet electrophoresis. The protein abundance for members of key signaling pathways was confirmed using western blotting.

Results

Atp8a1 deletion resulted in slight hyperleukocytosis, associated with the high proliferation of HSCs and BCR/ABL1 transformed leukemia stem cells (LSCs). Atp8a1 deletion increased the repopulation capability of HSCs with a competitive advantage in reconstitution assay. HSCs without Atp8a1 were more sensitive to 5-FU-induced apoptosis. Moreover, Atp8a1 deletion prevented HSC DNA damage and facilitated DNA repair processes. Genes involved in PI3K-AKT-mTORC1, DNA repair, and AP-1 complex signaling were enriched and elevated in HSCs with Atp8a1 deletion. Furthermore, Atp8a1 deletion caused decreased PTEN protein levels, resulting in the activation of PI3K-AKT-mTORC1 signaling, further increasing the activity of JNK/AP-1 signaling and YAP1 phosphorylation.

Conclusion

We identified the role of Atp8a1 on hematopoiesis and HSCs. Atp8a1 deletion resulted in the loss of phosphatidylserine asymmetry and intracellular signal transduction chaos.

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Data availability

The datasets used and analyzed during the current study are available from the corresponding author upon reasonable request. In addition, the raw sequence data reported in this paper have been deposited in the Genome Sequence Archive[28] in National Genomics Data Center[29], China National Center for Bioinformation / Beijing Institute of Genomics, Chinese Academy of Sciences (GSA: CRA009276), which is publicly accessible at https://ngdc.cncb.ac.cn/gsa.

This paper has not been previously published and is not under consideration by another journal. All authors have approved and agreed to submit the manuscript to this journal.

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Acknowledgements

The authors greatly appreciate the State Key Laboratory of Biotherapy & Collaborative Innovation Center for Biotherapy for their support, the staff of the core facility and the animal facility of the State Key Laboratory of Biotherapy and West China Hospital.

Funding

This work was supported by grants from the National Natural Science Foundation of China (82170114 to Y. Hu) and National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University (Z20201008 to Y. Hu), Guizhou Provincial Science & Technology Support Program (NO [2020]4Y061 to Y. Hu). National Natural Science Foundation of China (81802468 to LZh). Chengdu Science and Technology innovation project (2021-YF05-00800-SN to LZh).

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Author notes

  1. Li Zheng and Cong Pan these authors contributed equally to this work.

Authors and Affiliations

  1. Department of Thyroid Surgery, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, No.17, Ren-Min-Nan Road (Third Section), Chengdu, 610041, Sichuan, China

    Li Zheng, Cong Pan, Cailing Liang, Yunyu Feng, Wei He, Bochuan Wang, Qiang Qiu, Ning Li, Yuanyuan Sun, Huandi Qiu, Lingyun Zhou & Yiguo Hu

  2. The Sichuan Provincial Key Laboratory for Human Disease Gene Study, Center for Medical Genetics, Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, 610072, Sichuan, China

    Wanli Tian & Xianjun Zhu

  3. Institute of Life Science, eBond Pharmaceutical Technology Ltd., Chengdu, China

    Zirong Yang & Klarke M. Sample

  4. Center of Infectious Diseases, West China Hospital of Sichuan University, 37 GuoXue Lane, Chengdu, 610041, Sichuan Province, China

    Lingyun Zhou

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  1. Li Zheng
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Contributions

L.Z, C.P and Y.H conceived research ideas, designed experiments, analyzed data, and wrote the manuscript. L.Z., W.T. and C.L. performed experiments; Y.F and B.W performed the bioinformatics; H. Q, Q.Q, N.L; W.H. Y.S and Z. Y helped complete the experiments. L.Z, X.Z and K.S reviewed and edited the manuscript, discussed the results and commented on the manuscript. All authors have read and approved the article.

Corresponding authors

Correspondence to Lingyun Zhou, Xianjun Zhu or Yiguo Hu.

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All mouse studies were approved by the Institutional Animal Care and Use Committee (IACUC) at the Sichuan University. All animals were monitored for abnormal behavior to minimize pain and suffering. Animals were euthanized when excessive deterioration of health was noted.

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Zheng, L., Pan, C., Tian, W. et al. Atp8a1 deletion increases the proliferative activity of hematopoietic stem cells by impairing PTEN function. Cell Oncol. 46, 1069–1083 (2023). https://doi.org/10.1007/s13402-023-00797-7

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