CRISPR Screening of CAR T Cells and Cancer Stem Cells Reveals Critical Dependencies for Cell-Based Therapies

Cancer Discovery - Tập 11 Số 5 - Trang 1192-1211 - 2021
Dongrui Wang1, Briana C. Prager2,3,4, Ryan C. Gimple2,4,5, Brenda Aguilar1, Darya Alizadeh1, Hongzhen Tang6,7,8, Deguan Lv2,4, Renate Starr1, Alfonso Brito1, Qiulian Wu2,4, Leo J.Y. Kim2,4,5, Zhixin Qiu2,4, Lin Peng6,7,8, Michael H. Lorenzini2,4, Behnam Badie9, Stephen J. Forman1, Qi Xie6,7,8, Christine E. Brown1, Jeremy N. Rich10,2,4
11T Cell Therapeutics Research Labs, Cellular Immunotherapy Center, Department of Hematology and Hematopoietic Cell Transplantation, City of Hope, Duarte, California.
22Division of Regenerative Medicine, Department of Medicine, University of California, San Diego, San Diego, California.
33Cleveland Clinic Lerner College of Medicine at Cleveland Clinic and Case Western Reserve University, Cleveland, Ohio.
44Sanford Consortium for Regenerative Medicine, La Jolla, California.
55Department of Pathology, Case Western Reserve University, Cleveland, Ohio.
66Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China.
77Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China.
88Institute of Basic Medical Sciences, Westlake Institute for Advanced Study, Hangzhou, Zhejiang Province, China.
99Division of Neurosurgery, Department of Surgery, City of Hope, Duarte, California.
1010University of Pittsburgh Medical Center Hillman Cancer Center, Department of Neurology, University of Pittsburgh, Pittsburgh, Pennsylvania.

Tóm tắt

AbstractGlioblastoma (GBM) contains self-renewing GBM stem cells (GSC) potentially amenable to immunologic targeting, but chimeric antigen receptor (CAR) T-cell therapy has demonstrated limited clinical responses in GBM. Here, we interrogated molecular determinants of CAR-mediated GBM killing through whole-genome CRISPR screens in both CAR T cells and patient-derived GSCs. Screening of CAR T cells identified dependencies for effector functions, including TLE4 and IKZF2. Targeted knockout of these genes enhanced CAR antitumor efficacy. Bulk and single-cell RNA sequencing of edited CAR T cells revealed transcriptional profiles of superior effector function and inhibited exhaustion responses. Reciprocal screening of GSCs identified genes essential for susceptibility to CAR-mediated killing, including RELA and NPLOC4, the knockout of which altered tumor–immune signaling and increased responsiveness of CAR therapy. Overall, CRISPR screening of CAR T cells and GSCs discovered avenues for enhancing CAR therapeutic efficacy against GBM, with the potential to be extended to other solid tumors.Significance:Reciprocal CRISPR screening identified genes in both CAR T cells and tumor cells regulating the potency of CAR T-cell cytotoxicity, informing molecular targeting strategies to potentiate CAR T-cell antitumor efficacy and elucidate genetic modifications of tumor cells in combination with CAR T cells to advance immuno-oncotherapy.This article is highlighted in the In This Issue feature, p. 995

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Cancer Discovery

Tập 11 Số 5

1192-1211

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