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ACUTE MYELOID LEUKEMIA

Autophagy inhibition impairs leukemia stem cell function in FLT3-ITD AML but has antagonistic interactions with tyrosine kinase inhibition

Leukemia volume 36, pages 2621–2633 (2022)Cite this article

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Abstract

The FLT3-ITD mutation is associated with poor prognosis in acute myeloid leukemia (AML). FLT3 tyrosine kinase inhibitors (TKIs) demonstrate clinical efficacy but fail to target leukemia stem cells (LSC) and do not generate sustained responses. Autophagy is an important cellular stress response contributing to hematopoietic stem cells (HSC) maintenance and promoting leukemia development. Here we investigated the role of autophagy in regulating FLT3-ITD AML stem cell function and response to TKI treatment. We show that autophagy inhibition reduced quiescence and depleted repopulating potential of FLT3-ITD AML LSC, associated with mitochondrial accumulation and increased oxidative phosphorylation. However, TKI treatment reduced mitochondrial respiration and unexpectedly antagonized the effects of autophagy inhibition on LSC attrition. We further show that TKI-mediated targeting of AML LSC and committed progenitors was p53-dependent, and that autophagy inhibition enhanced p53 activity and increased TKI-mediated targeting of AML progenitors, but decreased p53 activity in LSC and reduced TKI-mediated LSC inhibition. These results provide new insights into the role of autophagy in differentially regulating AML stem and progenitor cells, reveal unexpected antagonistic effects of combined oncogenic tyrosine kinase inhibition and autophagy inhibition in AML LSC, and suggest an alternative approach to target AML LSC quiescence and regenerative potential.

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Fig. 1: Autophagy inhibition reduces growth of FLT3-ITD cells and enhances sensitivity to TKI treatment in vitro.
Fig. 2: Autophagy inhibition reduces human FLT3-ITD AML CD34+ cell growth and enhances TKI sensitivity in vitro.
Fig. 3: Autophagy inhibition enhances TKI-mediated targeting of primary human FLT3-ITD cells engrafted in immunodeficient mice.
Fig. 4: Autophagy inhibition enhances TKI-mediated inhibition of FLT3 ITD AML progenitors and increases AML stem cell cycling in a genetic mouse model.
Fig. 5: Autophagy inhibition reduces repopulating capacity of FLT3-ITD AML stem cells in a genetic mouse model.
Fig. 6: Enhanced mitochondrial respiration gene signatures in AML LSC following autophagy inhibition.
Fig. 7: Reduced mitochondrial respiration in AML stem cells with tyrosine kinase inhibition in combination with autophagy inhibition.
Fig. 8: Autophagy inhibition differentially modulates p53 activity in TKI-treated FLT3-ITD AML progenitor and stem cells.

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Data availability

RNA sequencing data is deposited in NCBI’s Gene Expression Omnibus (GEO) and is accessible through GEO Series accession number: GSE169750.

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Acknowledgements

This work was supported by NIH grant R01 CA172447 and R01 CA248794 to RB. We thank Drs Victor Darley-Usmar and Balu Chacko at the Comparative Mitochondrial Health Assessment Core for the cellular bioenergetics assays. We thank Vidya Sagar Hanumanthu for providing help with FACS sorting at the UAB Comprehensive Flow Cytometry Core; the UAB Animal resources Center for maintaining mice colonies; Maya Robinson and Amanda Mullens for obtaining and processing human samples; Mason Harris for assistance with animal studies. Immunofluorescence imaging was performed at the High-Resolution Imaging Facility at UAB.

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Authors and Affiliations

  1. Division of Hematology and Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

    Shaowei Qiu, Harish Kumar, Chengcheng Yan, Hui Li, Andrew J. Paterson, Nicholas R. Anderson, Jianbo He, Rui Lu, Robert S. Welner & Ravi Bhatia

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

    Shaowei Qiu

  3. Division of Cardiovascular Disease, University of Alabama at Birmingham, Birmingham, AL, USA

    Jing Yang & Min Xie

  4. Genomics Core Facility, University of Alabama at Birmingham, Birmingham, AL, USA

    David K. Crossman

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Conceptualization: SQ, RB; Methodology: SQ, HK, CY, AP, NA, and RB; Investigation: SQ, HK, NA, HL, JH, JY, MX, DC, RL, RW, and RB; Formal analysis: SQ, RB; Writing–Original Draft: SQ, RB; Writing–Review & Editing: RB, SQ, AP; Funding Acquisition: RB; Supervision: RB.

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Correspondence to Ravi Bhatia.

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Qiu, S., Kumar, H., Yan, C. et al. Autophagy inhibition impairs leukemia stem cell function in FLT3-ITD AML but has antagonistic interactions with tyrosine kinase inhibition. Leukemia 36, 2621–2633 (2022). https://doi.org/10.1038/s41375-022-01719-6

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