To identify molecules/pathways governing Venetoclax (VEN) sensitivity, we performed genome-wide CRISPR/Cas9 screens using a mouse AML line insensitive to VEN-induced mitochondrial apoptosis. Levels of sgRNAs targeting March5, Ube2j2 or Ube2k significantly decreased upon VEN treatment, suggesting synthetic lethal interaction. Depletion of either Ube2j2 or Ube2k sensitized AML cells to VEN only in the presence of March5, suggesting coordinate function of the E2s Ube2j2 and Ube2k with the E3 ligase March5. We next performed CRISPR screens using March5 knockout cells and identified Noxa as a key March5 substrate. Mechanistically, Bax released from Bcl2 upon VEN treatment was entrapped by Mcl1 and Bcl-XL and failed to induce apoptosis in March5 intact AML cells. By contrast, in March5 knockout cells, liberated Bax did not bind to Mcl1, as Noxa likely occupied Mcl1 BH3-binding grooves and efficiently induced mitochondrial apoptosis. We reveal molecular mechanisms underlying AML cell-intrinsic VEN resistance and suggest a novel means to sensitize AML cells to VEN.
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The processed data of CRISPR/Cas9 screens are available as Supplementary tables. Raw data will be provided upon request.
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We thank the members of the Department of Medicine and Biosystemic Science and the Division of Precision Medicine at Kyushu University for assistance and helpful discussion, and Elise Lamar for critical reading of the manuscript. This work is supported in part by a Grant-in-Aid for Young Scientists (18K16089)(to YS), a Grant-in-Aid for Young Scientists (19K17859), a Research Grant from the KANAE Foundation, the MSD Life Science Foundation, the Yasuda Medical Foundation, the Mochida Memorial Foundation for Medical and Pharmaceutical Research, the Shinnihon Foundation of Advanced Medical Treatment Research, the Takeda Science Foundation (to TY), a Grant-in-Aid for Scientific Research (C)(21K08429), the Shinnihon Foundation of Advanced Medical Treatment Research, the Takeda Science Foundation (to KM), a Grant-in-Aid for Scientific Research (B)(22494899)(to YK), a Grant-in-Aid for Scientific Research (C)(19K05544) (to T Masuda), a Grant-in-Aid for Scientific Research (S)(16H06391) (to KA), a Grant-in-Aid for Scientific Research (A) (17H01567, 20H00540), an AMED under grant number 18063889 and a Grant-in-Aid for Scientific Research (S)(20H05699)(to T Maeda).
The authors declare no competing interests.
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Nakao, F., Setoguchi, K., Semba, Y. et al. Targeting a mitochondrial E3 ubiquitin ligase complex to overcome AML cell-intrinsic Venetoclax resistance. Leukemia 37, 1028–1038 (2023). https://doi.org/10.1038/s41375-023-01879-z