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This study aims to explore the molecular determinants of cancer cell sensitivity to CAR\u2010T cell\u2010mediated killing and to provide a better understanding of the underlying mechanisms and potential modulation to improve clinical efficacy.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec xml:lang=\"en\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p xml:lang=\"en\">The human whole\u2010genome CRISPR\/Cas9\u2010based knockout screening was conducted to identify key genes that enable cancer cells to evade CD19 CAR\u2010T\u2010cell\u2010mediated killing. The in vitro cytotoxicity assays and evaluation of tumor tissue and bone marrow specimens were further conducted to confirm the role of the key genes in cancer cell susceptibility to CAR\u2010T cells. In addition, the specific mechanisms influencing CAR\u2010T cell\u2010mediated cancer clearance were elucidated in mouse and cellular models.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec xml:lang=\"en\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p xml:lang=\"en\">\n                      The CRISPR\/Cas9\u2010based knockout screening showed that the enrichment of autophagy\u2010related genes (\n                      <jats:italic toggle=\"yes\">ATG3<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">BECN1<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">RB1CC1<\/jats:italic>\n                      ) provided protection of cancer cells from CD19 CAR\u2010T cell\u2010mediated cytotoxicity. These findings were further validated by in vitro cytotoxicity assays in cells with genetic and pharmacological inhibition of autophagy. Notably, higher expression of the three autophagy\u2010related proteins in tumor samples was correlated with poorer responsiveness and worse survival in patients with relapsed\/refractory B\u2010cell lymphoma after CD19 CAR\u2010T therapy. Bulk RNA sequencing analysis of bone marrow samples from B\u2010cell leukemia patients also suggested the clinical relevance of autophagy to the therapeutic response and relapse after CD19 CAR\u2010T cell therapy. Pharmacological inhibition of autophagy and knockout of RB1CC1 could dramatically sensitize tumor cells to CD19 CAR\u2010T cell\u2010mediated killing in mouse models of both B\u2010cell leukemia and lymphoma. Moreover, our study revealed that cancer\u2010intrinsic autophagy mediates evasion of CAR\u2010T cells via the TNF\u2010\u03b1\u2010TNFR1 axis\u2010mediated apoptosis and STAT1\/IRF1\u2010induced chemokine signaling activation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec xml:lang=\"en\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p xml:lang=\"en\">These findings confirm that autophagy signaling in B\u2010cell malignancies is essential for the effective cytotoxic function of CAR\u2010T cells and thereby pave the way for the development of autophagy\u2010targeting strategies to improve the clinical efficacy of CAR\u2010T cell immunotherapy.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/cac2.12525","type":"journal-article","created":{"date-parts":[[2024,2,26]],"date-time":"2024-02-26T11:40:45Z","timestamp":1708947645000},"page":"408-432","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Targeting autophagy overcomes cancer\u2010intrinsic resistance to CAR\u2010T immunotherapy in B\u2010cell malignancies"],"prefix":"10.34133","volume":"44","author":[{"given":"Lu","family":"Tang","sequence":"first","affiliation":[{"name":"Union Hospital","place":["Wuhan, P. 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