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Molecular targets for therapy

Sensitizing acute myeloid leukemia cells to induced differentiation by inhibiting the RIP1/RIP3 pathway

Leukemia volume 31, pages 1154–1165 (2017)Cite this article

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Abstract

Tumor necrosis factor-α (TNF-α)-induced RIP1/RIP3 (receptor-interacting protein kinase 1/receptor-interacting protein kinase 3)-mediated necroptosis has been proposed as an alternative strategy for treating apoptosis-resistant leukemia. However, we found that most acute myeloid leukemia (AML) cells, especially M4 and M5 subtypes, produce TNF and show basal level activation of RIP1/RIP3/MLKL signaling, yet do not undergo necroptosis. TNF, through RIP1/RIP3 signaling, prevents degradation of SOCS1, a key negative regulator of interferon-γ (IFN-γ) signaling. Using both pharmacologic and genetic assays, we show here that inactivation of RIP1/RIP3 resulted in reduction of SOCS1 protein levels and partial differentiation of AML cells. AML cells with inactivated RIP1/RIP3 signaling show increased sensitivity to IFN-γ-induced differentiation. RIP1/RIP3 inactivation combined with IFN-γ treatment significantly attenuated the clonogenic capacity of both primary AML cells and AML cell lines. This combination treatment also compromised the leukemogenic ability of murine AML cells in vivo. Our studies suggest that inhibition of RIP1/RIP3-mediated necroptotic signaling might be a novel strategy for the treatment of AML when combined with other differentiation inducers.

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Acknowledgements

We thank the staff of the Department of Comparative Medicine of Loyola University Medical Center for animal care services, as well as Drs Manuel Diaz, Nancy Zeleznik-Le, Andrew Dingwall and Wei Qiu for their ongoing professional collaboration and scientific suggestions and discussions that have improved the present studies. We also thank the members of Dr Wei Qiu’s and Dr Stephanie Watkins’s laboratories for imaging assistance. We appreciate laboratory support from Patience Oladeinde, Danielle Howard and Emma Yao, and FACS sorting and analysis assistance from Patricia Simms, Ashley Hess, Shwetha Ravichandran and Veronica Volgina. This work was supported by NIH (Grants R01-HL095896 and R21-CA181970 to JZ through Loyola University Chicago) and the Leukemia Research Foundation New Investigator Award (8th Annual George Richard Memorial Grant to JX). JX was also supported in part by a grant from the Muscular Dystrophy Association (MDA202906). AV was supported by NRSA F31 Fellowship (F31CA17417). JL and Jun Z were supported by National Basic Research Program of China (Project 2013CB966800), the Program for Basic Research of Shanghai Municipal Science and Technology Commission (Grant No. 13JC1406403).

Author contributions

JX designed the experiments, analyzed the data and drafted the manuscript; Jiwang Z supervised the overall research, designed the experiments, analyzed the data and edited the manuscript; JX, JL, Jun Z, RS, DY, AV, AN, GN, RG, JC, JP, WW, ZX, YC, PB, HC, Jiang Z and Jianke Z collectively contributed to data collection and interpretation of the results; JC provided microarray data; MMP, PCK, SN and AK provided study samples, clinical data, helped to write and revised the manuscript. PB helped to write, and also edited and refined the manuscript.

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Authors and Affiliations

  1. Oncology Institute, Cardinal Bernardin Cancer Center, Loyola University Medical Center, Maywood, IL, USA

    J Xin, D You, P Breslin, W Wei, J Cannova, A Volk, R Gutierrez, Y Xiao, A Ni, G Ng, R Schmidt, Z Xia, J Pan, P C Kuo, S Nand, A R Kini & J Zhang

  2. Research and Development Service, Hines VA Hospital, Hines, IL, USA

    J Xin

  3. Department of Molecular Pharmacology and Therapeutics, Loyola University Medical Center, Maywood, IL, USA

    J Xin

  4. Department of Biology, Loyola University Chicago, Chicago, IL, USA

    P Breslin

  5. Department of Molecular/Cellular Physiology, Loyola University Medical Center, Maywood, IL, USA

    P Breslin

  6. Department of Biology, College of Life and Environment Science, Shanghai Normal University, Shanghai, People’s Republic of China

    J Li & J Zhang

  7. Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Jilin University, Changchun, China

    Y Xiao

  8. Department of Pathology, Loyola University Medical Center, Maywood, IL, USA

    H Chen, A R Kini & J Zhang

  9. Department of Medicine, Loyola University Medical Center, Maywood, IL, USA

    M M Patel & S Nand

  10. Department of Microbiology and Immunology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA, USA

    J Zhang

  11. Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH, USA

    J Chen

  12. State Key Laboratory for Medical Genomics and Shanghai Institute of Hematology and Collaborative Innovation Center of Hematology, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, People’s Republic of China

    J Zhu

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  1. J Xin
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Correspondence to J Xin or J Zhang.

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Xin, J., You, D., Breslin, P. et al. Sensitizing acute myeloid leukemia cells to induced differentiation by inhibiting the RIP1/RIP3 pathway. Leukemia 31, 1154–1165 (2017). https://doi.org/10.1038/leu.2016.287

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