Besnik Qeriqi

Supplementary Figure 1. Activation status of anti-U5 snRNP200 CAR T cells co-cultured with AML cells in vitro. Supplementary Figure 2. Efficacy of IL-18 armored anti-U5 snRNP200 CAR T cells and their dependency on IFNg secretion. Supplementary Figure 3. Efficacy of IL-18 armored anti-U5 snRNP200 CAR T cells in AMLPDX models. Supplementary Figure 4. Evaluation of IL-18 armored U5 snRNP200 CAR T cells in pediatric AML PDXs. Supplementary Figure 5. Safety assessment of anti-U5 snRNP200 CAR T cells. Supplementary Figure 6. Efficacy of mouse IL-18 secreting U5 snRNP200 CAR T cells in a syngeneic mouse AML model. Supplementary Figure 7. UMAPs of bone marrow cells annotated by cell type and marker gene expression in cell type. Supplementary Figure 8. Clonal expansion of endogenous T cells in mice treated with IL18 secreting anti-U5 snRNP200 CAR T cells. Supplementary Figure 9. Evaluation of mechanism underlying AML re-challenge in immunocompetent mice following AML treatment with U5 snRNP200 CAR T cells. Supplementary Figure 10. Surface expression of U5 snRNP200 and efficacy of anti-U5 snRNP200 CAR T cells in B-ALL. Supplementary Figure 11. Anti-leukemic activity of U5 snRNP200 against B-ALL in vivo.

Supplementary Table S1. Clinical and genetic characteristics of AML and B-ALL patients for PDX models. Supplementary Table S2. TCR sequences of bone marrow T cells in syngeneic AML mouse model. Supplementary Table S3. Clinical and genetic characteristics of B-ALL patients for evaluation of surface U5 snRNP200 expression.

Developing chimeric antigen receptor (CAR) T cells for acute myeloid leukemia (AML) has been challenging due to a lack of known AML-associated antigens that spare normal hematopoietic precursor cells. Here we reasoned that donor autoantibodies from AML recipients cured following allogeneic transplant and responsible for graft-versus-leukemia effect could be engineered to create effective CAR-T cells. We generated CAR-T cells against one such antigen - U5 snRNP200, an RNA helicase localized to the AML cell surface and absent from normal hematopoietic precursors. Anti-U5 snRNP200 CAR-T cells were effective in human and syngeneic models of AML as well as B-cell acute lymphoblastic leukemia (B-ALL), a setting where surface U5 snRNP200 is also present. Armoring CAR-T cells with IL-18 led to antigen gain on AML, durable remission, and protection from AML rechallenge. These data thereby identify a CAR-T cell platform which addresses prior limitations in tumor-selectivity and safety for patients with acute leukemias.

RW7213 cell lines sequencing analysis

C106 cell lines single cell sequencing analysis

Patients Progression Cell Free DNA data

Generation of acquired resistance PDX model

Patients Longitudinal Cell Free DNA data

Characterization of acquired resistance in vitro models