학술논문
Leukemia-intrinsic determinants of CAR-T response revealed by iterative in vivo genome-wide CRISPR screening
Document Type
article
Author
Azucena Ramos; Catherine E. Koch; Yunpeng Liu-Lupo; Riley D. Hellinger; Taeyoon Kyung; Keene L. Abbott; Julia Fröse; Daniel Goulet; Khloe S. Gordon; Keith P. Eidell; Paul Leclerc; Charles A. Whittaker; Rebecca C. Larson; Audrey J. Muscato; Kathleen B. Yates; Juan Dubrot; John G. Doench; Aviv Regev; Matthew G. Vander Heiden; Marcela V. Maus; Robert T. Manguso; Michael E. Birnbaum; Michael T. Hemann
Source
Nature Communications, Vol 14, Iss 1, Pp 1-21 (2023)
Subject
Language
English
ISSN
2041-1723
Abstract
Abstract CAR-T therapy is a promising, novel treatment modality for B-cell malignancies and yet many patients relapse through a variety of means, including loss of CAR-T cells and antigen escape. To investigate leukemia-intrinsic CAR-T resistance mechanisms, we performed genome-wide CRISPR-Cas9 loss-of-function screens in an immunocompetent murine model of B-cell acute lymphoblastic leukemia (B-ALL) utilizing a modular guide RNA library. We identified IFNγR/JAK/STAT signaling and components of antigen processing and presentation pathway as key mediators of resistance to CAR-T therapy in vivo; intriguingly, loss of this pathway yielded the opposite effect in vitro (sensitized leukemia to CAR-T cells). Transcriptional characterization of this model demonstrated upregulation of these pathways in tumors relapsed after CAR-T treatment, and functional studies showed a surprising role for natural killer (NK) cells in engaging this resistance program. Finally, examination of data from B-ALL patients treated with CAR-T revealed an association between poor outcomes and increased expression of JAK/STAT and MHC-I in leukemia cells. Overall, our data identify an unexpected mechanism of resistance to CAR-T therapy in which tumor cell interaction with the in vivo tumor microenvironment, including NK cells, induces expression of an adaptive, therapy-induced, T-cell resistance program in tumor cells.