Abstract 879: Functional and single-cell assessment of CRISPR-modified CAR-T cells from NSCLC patients and healthy donors

Abstract

Abstract Cancer patients that have undergone multiple lines of treatment can have dysfunctional immune systems. In both solid tumors and hematological malignancies, T cells have been reported to be in various states of dysfunction including senescence or exhaustion. In order to assess the potency of CAR-T cells derived from cancer patients, we created CAR-T cells targeting CD70 from non-small cell lung cancer (NSCLC) patients as well as healthy donors. Anti-CD70 CAR-T cells were created by using CRISPR/Cas9 to insert the CAR cassette into a knocked-out TRAC locus. The TRAC knock-out eliminates any TCR-mediated effector functions. CAR-T preparations from either NSCLC patients or healthy donors were assessed in vitro for T cell effector functions including cell killing, cytokine secretion and expansion in the presence of lung tumor cells expressing antigen. CAR-T cells from NSCLC patients and healthy donors were also assessed for their ability to control human lung cancer-derived tumors in immunocompromised mice. Following these functional assessments, the heterogeneity of CAR-T preparations derived from different sources was characterized further by performing single-cell RNA sequencing analysis and single-cell protein analysis (CITE-Seq and single cell cytokine secretion). A superior understanding of CAR-T cell heterogeneity and donor variability could aid in producing more efficacious drug products for cancer. Citation Format: Zinkal Padalia, Andrew Dunn, Konstantinos Karagiannis, Brigid McEwan, Meghna Kuppuraju, Jason Sagert, Sushant Karnik, Mary-Lee Dequeant, Jonathan Terrett, Demetrios Kalaitzidis. Functional and single-cell assessment of CRISPR-modified CAR-T cells from NSCLC patients and healthy donors [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 879.