Delphine Gitenay

Supplementary Table S2 provides the sequences of the primers used for ATAC-Seq library preparation

Supplementary Figure S1 shows that TAK-981 induces CD69 and LFA-1 but not does not affect DNAM1 and TIGIT expression at the surface of NK cells

Supplementary Figure S2 shows that SUMOylation regulates IFN-I pathway genes enhancers activity and accessibility

Supplementary Table S5 provides the raw data for live cell cytotoxicity experiments using Incucyte

Supplementary Table S6 provides the clinical characteristics of all patient samples used in this study

Supplementary Table S4 provides the sequence of the gRNA used to knock out MDA5, IRF1, IRF3, IRF7 in THP1 cells

Supplementary Table S3 provides the sequence of the primers used for RT-qPCR

NK cells can play a significant role in the antitumoral immune response. In patients with acute myeloid leukemia (AML), NK cells are, however, often found in low numbers and exhibit poor activity, contributing to leukemic progression. Allogenic NK cells are emerging as promising cellular therapies for hematologic cancer treatment. New strategies are however required to both reactivate NK cells in patients with AML and enhance the antitumor activity of transplanted NK cells. In this study, we demonstrate that targeting SUMOylation, a protein posttranslational modification, activates NK cells from both healthy donors and patients with AML. Subasumstat (TAK-981), a first-in-class inhibitor of SUMOylation used in phase I/II clinical trials, enhances NK cell degranulation, secretion of inflammatory cytokines (IFN-γ, TNF-α, and FasL), and cytotoxicity against AML cells. In vivo, TAK-981 improves the anti-leukemic efficacy of ex vivo expanded cord blood NK cells in leukemia-bearing mice. One early effect of TAK-981 is to specifically increase the accessibility and activation of cis-regulatory regions of IFN-I pathway genes and induce their transcription. TAK-981–induced secretion of IFN-β, mostly by NK cells and myeloid cells, is required for NK cell activation. Surprisingly, IFNB1 induction does not require its best-characterized activators MDA5, cGas, and IFN response factor-1, -3, and -7. Altogether, this suggests that targeting SUMOylation activates a noncanonical IFN-I pathway, which enhances the anti-leukemic potential of NK cells.

Supplementary Table S1 provides information on the antibodies used for flow cytometry

Supplementary Table S7 provides a differential analysis of RNA-Seq results and Gene Set Enrichment Analysis (GSEA)