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STEM CELL BIOLOGY

Transcription factor Nkx2-3 maintains the self-renewal of hematopoietic stem cells by regulating mitophagy

Leukemia volume 37, pages 1361–1374 (2023)Cite this article

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Abstract

Hematopoietic stem cells (HSCs) reside at the top of the hematopoietic hierarchy, exhibiting a unique capacity to self-renew and differentiate into all blood cells throughout the lifetime. However, how to prevent HSC exhaustion during long-term hematopoietic output is not fully understood. Here, we show that the homeobox transcription factor Nkx2-3 is required for HSC self-renewal by preserving metabolic fitness. We found that Nkx2-3 is preferentially expressed in HSCs with excessive regenerative potential. Mice with conditional deletion of Nkx2-3 displayed a reduced HSC pool and long-term repopulating capacity as well as increased sensitivity to irradiation and 5-flurouracil treatment due to impaired HSC quiescence. In contrast, overexpression of Nkx2-3 improved HSC function both in vitro and in vivo. Furthermore, mechanistic studies revealed that Nkx2-3 can directly control the transcription of the critical mitophagy regulator ULK1, which is essential for sustaining metabolic homeostasis in HSCs by clearing activated mitochondria. More importantly, a similar regulatory role of NKX2-3 was observed in human cord blood-derived HSCs. In conclusion, our data demonstrate an important role of the Nkx2-3/ULK1/mitophagy axis in regulating the self-renewal of HSCs, therefore providing a promising strategy to improve the function of HSCs in the clinic.

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Fig. 1: Nkx2-3 is preferentially expressed in HSCs with great regenerative potential.
Fig. 2: Conditional knockout of Nkx2-3 results in a reduction in the HSC pool in mice.
Fig. 3: Deletion of Nkx2-3 intrinsically compromises the long-term repopulation ability of HSCs.
Fig. 4: Nkx2-3 deficiency significantly impairs the quiescence of HSCs.
Fig. 5: Ablation of Nkx2-3 leads to the accumulation of activated mitochondria in HSCs due to attenuated mitophagy.
Fig. 6: Nkx2-3 regulates mitophagy in HSCs by transcriptionally controlling the expression of Ulk1.
Fig. 7: The NKX2-3/ULK1/mitophagy axis maintains the self-renewal of human HSCs.

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Data availability

The raw data of scRNA-seq were deposited in the NCBI Gene Expression Omnibus (GEO) database (no. GSE221907). The raw data of bulk RNA-seq were deposited in the NCBI Sequence Read Archive (SRA) database (no. PRJNA891790).

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Acknowledgements

We thank Yang Liu for technical support in flow cytometry, Liting Wang for technical support in immunofluorescence microscopy, and Quanfang Wei for technical assistance in transmission electron microscopy. This work was supported by grants from the National Natural Science Foundation of China (No. 81930090, 82203974, 81725019), Project funded by China Postdoctoral Science Foundation (No. 2022M723867), Postdoctoral Innovative Talent Support Program of China (No. BX20220398), Postdoctoral Innovative Talent Support Program of Chongqing (No. CQBX2021017) and the Scientific Research Project of PLA (No. BWS21J022).

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  1. These authors contributed equally: Mengjia Hu, Naicheng Chen, Mo Chen.

Authors and Affiliations

  1. State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing, 400038, China

    Mengjia Hu, Naicheng Chen, Mo Chen, Fang Chen, Yukai Lu, Yang Xu, Lijing Yang, Hao Zeng, Mingqiang Shen, Shilei Chen, Fengchao Wang, Song Wang & Junping Wang

  2. Chinese PLA Center for Disease Control and Prevention, Beijing, 100071, China

    Mengjia Hu

  3. Department of Obstetrics and Gynecology, Liangping District Maternal and Child Health Care Hospital, Chongqing, 405200, China

    Xuehong Chen

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  1. Mengjia Hu
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Contributions

MH, NC and MC designed the study, performed experiments, analyzed data and wrote the paper. FC, YL and YX performed experiments and analyzed data. LY and HZ participated in some animal experiments. MS and XC participated in some in vitro experiments. SC and FW participated in the initial experimental design and discussed the manuscript. JW and SW conceived and supervised the study, and revised the manuscript.

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Correspondence to Song Wang or Junping Wang.

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Hu, M., Chen, N., Chen, M. et al. Transcription factor Nkx2-3 maintains the self-renewal of hematopoietic stem cells by regulating mitophagy. Leukemia 37, 1361–1374 (2023). https://doi.org/10.1038/s41375-023-01907-y

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