Non-viral precision T cell receptor replacement for personalized cell therapy.
The T cell receptor (TCR) provides the fine specificity of T cells to recognize mutations in cancer cells 1-3. We developed a clinical-grade approach based on CRISPR/Cas9 non-viral precision genome editing to simultaneously knock-out the two endogenous TCR genes, TCRα (TRAC) and TCRβ (TRBC), and insert in the TRAC locus the two chains of a neoantigen-specific TCR (neoTCR), isolated from the patient's own circulating T cells using a personalized library of soluble predicted neoantigen-HLA capture reagents. Sixteen patients with refractory solid cancers received up to three distinct neoTCR-transgenic cell products, each expressing a patient-specific neoTCR, in a cell dose-escalation, first-in-human phase 1 clinical trial (NCT03970382). One patient had grade 1 cytokine release syndrome, and one grade 3 encephalitis. All had the expected side effects from the lymphodepleting chemotherapy. Five patients had stable disease, and the other 11 had disease progression as best response on therapy. NeoTCR-transgenic T cells were detected in tumour biopsies post-infusion at frequencies higher than the native TCRs pre-infusion. This study demonstrates the feasibility of isolating and cloning multiple TCRs recognizing mutational neoantigens, the simultaneous knock-out of the endogenous TCR and knock-in of the neoTCRs using single-step, non-viral precision genome editing, the manufacturing of neoTCR engineered T cells at clinical grade, the safety of infusing up to three gene edited neoTCR T cell products, and the ability of the transgenic T cells to traffic to the patients' tumours.
Institute for Systems Biology
Foy, Susan P; Jacoby, Kyle; Bota, Daniela A; Hunter, Theresa; Pan, Zheng; Stawiski, Eric; Ma, Yan; Lu, William; Peng, Songming; Wang, Clifford L; Yuen, Benjamin; Dalmas, Olivier; Heeringa, Katharine; Sennino, Barbara; Conroy, Andy; Bethune, Michael T; Mende, Ines; White, William; Kukreja, Monica; Gunturu, Swetha; Humphrey, Emily; Hussaini, Adeel; An, Duo; Litterman, Adam J; Quach, Boi Bryant; Ng, Alphonsus H C; Lu, Yue; Smith, Chad; Campbell, Katie M; Anaya, Daniel; Skrdlant, Lindsey; Huang, Eva Yi-Hsuan; Mendoza, Ventura; Mathur, Jyoti; Dengler, Luke; Purandare, Bhamini; Moot, Robert; Yi, Michael C; Funke, Roel; Sibley, Alison; Stallings-Schmitt, Todd; Oh, David Y; Chmielowski, Bartosz; Abedi, Mehrdad; Yuan, Yuan; Sosman, Jeffrey A; Lee, Sylvia M; Schoenfeld, Adam J; Baltimore, David; Heath, James R; Franzusoff, Alex; Ribas, Antoni; Rao, Arati V; and Mandl, Stefanie J, "Non-viral precision T cell receptor replacement for personalized cell therapy." (2022). Articles, Abstracts, and Reports. 6700.