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ACUTE MYELOID LEUKEMIA

Microenvironmental CXCL12 deletion enhances Flt3-ITD acute myeloid leukemia stem cell response to therapy by reducing p38 MAPK signaling

Leukemia volume 37, pages 560–570 (2023)Cite this article

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Abstract

Fms-like tyrosine kinase 3 (Flt3) tyrosine kinase inhibitors (Flt3-TKI) have improved outcomes for patients with Flt3-mutated acute myeloid leukemia (AML) but are limited by resistance and relapse, indicating persistence of leukemia stem cells (LSC). Here utilizing a Flt3-internal tandem duplication (Flt3-ITD) and Tet2-deleted AML genetic mouse model we determined that FLT3-ITD AML LSC were enriched within the primitive ST-HSC population. FLT3-ITD LSC showed increased expression of the CXCL12 receptor CXCR4. CXCL12-abundant reticular (CAR) cells were increased in Flt3-ITD AML marrow. CXCL12 deletion from the microenvironment enhanced targeting of AML cells by Flt3-TKI plus chemotherapy treatment, including enhanced LSC targeting. Both treatment and CXCL12 deletion partially reduced p38 mitogen-activated protein kinase (p38) signaling in AML cells and further reduction was seen after treatment in CXCL12 deleted mice. p38 inhibition reduced CXCL12-dependent and -independent maintenance of both murine and human Flt3-ITD AML LSC by MSC and enhanced their sensitivity to treatment. p38 inhibition in combination with chemotherapy plus TKI treatment leads to greater depletion of Flt3-ITD AML LSC compared with CXCL12 deletion. Our studies support roles for CXCL12 and p38 signaling in microenvironmental protection of AML LSC and provide a rationale for inhibiting p38 signaling to enhance Flt3-ITD AML targeting.

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Fig. 1: AML development in Flt3-ITD TET2 flox/flox Mx-1 cre mice.
Fig. 2: Flt3-ITD TET2-KO LSC reside in primitive phenotypic HSPC compartments.
Fig. 3: Flt3-ITD AML alters the BM microenvironment and increases CXCL12 expression in CAR cells.
Fig. 4: CXCL12 KO reduces AML disease burden.
Fig. 5: Effect of CXCL12 KO on quizartinib response.
Fig. 6: CXCL12 KO enhances response to combination chemotherapy plus quizartinib.
Fig. 7: Changes in gene expression in leukemia stem cells following CXCL12 KO and combination chemotherapy plus TKI.
Fig. 8: p38 signaling contributes to AML LSK resistance to chemo + TKI.

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Data availability

The datasets generated during the current study are available from the corresponding author on reasonable request. RNAseq data is deposited in NCBI’s Gene Expression Omnibus (GEO) and is accessible through GEO Series accession number: GSE212428.

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Acknowledgements

This work was supported by National Institutes of Health grant R01 number CA1772447 to RB. NA was supported in part by UAB Cell and Molecular Biology T32 Training Grant T32GM008111. The authors would like to thank: AML patients at UAB for the generous donation of their tissue samples; Drs. Douglas Hurst, Amjad Javed and Christopher Klug for helpful scientific discussions and comments; Maya Robinson and Amanda Mullen-Clem for procurement and processing of human samples. HudsonAlpha Institute for Biotechnology Sequencing Facility, Huntsville, Alabama for performing RNA sequencing; the UAB Comprehensive Flow Cytometry Core for providing help with FACS sorting; and the UAB Animal Resource Program for maintaining mouse colonies. NRA is a PhD candidate at the University of Alabama at Birmingham. This work is submitted in partial fulfillment of the requirement for the PhD.

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Authors and Affiliations

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

    Nicholas R. Anderson, Vipul Sheth, Hui Li, Mason W. Harris, Shaowei Qiu, Harish Kumar, 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. Department of Genetics, University of Alabama at Birmingham, Birmingham, AL, USA

    David K. Crossman

  4. Division of Experimental Hematology & Cancer Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA

    Puneet Agarwal

  5. Laboratory of Stem Cell Biology & Developmental Immunology, Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan

    Takashi Nagasawa

  6. Division of Endocrinology, Diabetes, and Metabolism, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA

    Andrew J. Paterson

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Contributions

Conceptualization: NRA, AJP, and RB. Methodology: NRA, VS, PA, SQ, RSW. Investigation: NRA, VS, MH, HL, HK, and SQ. Formal analysis: NRA, VS and DC. Resources: TN. Writing-Original draft Editing: NRA, VS and RB. Writing- Review and Editing: all authors. Funding acquisition: RB.

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Correspondence to Ravi Bhatia.

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Anderson, N.R., Sheth, V., Li, H. et al. Microenvironmental CXCL12 deletion enhances Flt3-ITD acute myeloid leukemia stem cell response to therapy by reducing p38 MAPK signaling. Leukemia 37, 560–570 (2023). https://doi.org/10.1038/s41375-022-01798-5

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