Supplementary Figures S1-S33 from SOCS1 Protects Acute Myeloid Leukemia against Allogeneic T Cell–Mediated Cytotoxicity

Supplementary Figure S1. Genome-wide CRISPR screening method and validation measures. Supplementary Figure S2. SOCS1-silenced AML cell line validation. Supplementary Figure S3. SOCS1-silenced primary AML blast validation. Supplementary Figure S4. SOCS1-overexpressed AML cell line validation. Supplementary Figure S5. SOCS1OE OCI-AML2 cells are resistant to anti-CD4 CAR (CAR4)-T cell killing. Supplementary Figure S6. SOCS1-overexpressed and control OCI-AML2 cells are killed similarly by DNTs in a short-term assay. Supplementary Figure S7. Overexpression or knockdown of SOCS1 does not influence AML sensitivity to daunorubicin or venetoclax. Supplementary Figure S8. SOCS3 and CISH do not correlate with AML susceptibility to DNTs, and CRISPR screening only identifies SOCS1 among other SOCS family members as a significant gene that alters AML susceptibility to DNT killing. Supplementary Figure S9. JAK1-silenced AML cell line validation. Supplementary Figure S10. JAK1-silenced MV4-11 in vivo model schematic. Supplementary Figure S11. JAK1 expression does not correlate with AML susceptibility to DNTs or patient survival. Supplementary Figure S12. Gating strategy to detect changes in pSTAT1 in AML following DNT co-culture. Supplementary Figure S13. IFNγ upregulates pSTAT1 in a JAK1-dependent manner. Supplementary Figure S14. JAK1-silencing prevents rIFNγ-mediated sensitization of AML cells. Supplementary Figure S15. IFNγ has minimal effect on viability of SOCS1-overexpressed and knockdown AML cells. Supplementary Figure S16. SOCS1-overexpression and JAK1-silencing does not significantly alter DNT secretion of IFNγ. Supplementary Figure S17. TNFα and IFNα sensitization effects are augmented by SOCS1 expression in AML. Supplementary Figure S18. SOCS1 overexpression reduces IFNγ-induced ICAM-1 upregulation. Supplementary Figure S19. SOCS1OE OCI-AML2 cells are resistant to ICAM-1 induction by CAR4-T cells. Supplementary Figure S20. JAK1-silencing abrogates DNT-mediated ICAM-1 upregulation. Supplementary Figure S21. Validation of ICAM-1KO OCI-AML2 cell line. Supplementary Figure S22. AML viability after antibody and IFNγ treatment. Supplementary Figure S23. Blocking LFA-1 prevents the AML sensitization effect of SOCS1KD and IFNγ treatment to DNT killing. Supplementary Figure S24. SOCS1 correlation with survival in various patient cohorts. Supplementary Figure S25. ICAM-1 and IFNγ levels do not correlate with patient survival, and inducible ICAM-1 expression is transient. Supplementary Figure S26. In vitro growth of SOCS1 knockdown and overexpressed cells. Supplementary Figure S27. AML patients experiencing complete remission or primary induction failure after chemotherapy have similar SOCS1 levels. Supplementary Figure S28. Leukemic stem cell scores of annotated cellular clusters from Lasry et al. dataset. Supplementary Figure S29. Inferred up- and downregulated interactions between HSCs and effector cells among SOCS1low cells. Supplementary Figure S30. Risk profile of SOCS1low patients of the PMCC cohort. Supplementary Figure S31. Representative gating strategy of in vitro cytotoxicity assays against AML cell lines. Supplementary Figure S32. Representative gating strategy for primary AML samples. Supplementary Figure S33. Representative gating strategy of in vivo AML xenograft model.