Hypoimmune anti-CD19 chimeric antigen receptor T cells provide lasting tumor control in fully immunocompetent allogeneic humanized mice

• Published in: Nature communications Vol. 14; no. 1; p. 2020
• Main Authors: Hu, Xiaomeng, Manner, Karl, DeJesus, Rowena, White, Kathy, Gattis, Corie, Ngo, Priscilla, Bandoro, Christopher, Tham, Eleonore, Chu, Elaine Y., Young, Chi, Wells, Frank, Basco, Ronald, Friera, Annabelle, Kangeyan, Divy, Beauchesne, Pascal, Dowdle, William E., Deuse, Tobias, Fry, Terry J., Foster, Aaron E., Schrepfer, Sonja
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 10.04.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 13/106; 13/109; 13/31; 14/5; 38; 38/5; 59; 59/5; 631/250/2152/2153; 64; 64/60; 692/4028/67; Animals; Antigens; Cancer; CD19 antigen; CD47 Antigen; Chimeric antigen receptors; CIITA protein; CRISPR; Effectiveness; Hematopoietic Stem Cell Transplantation; Humanities and Social Sciences; Humans; Immune clearance; Immune system; Immunocompetence; Immunodeficiency; Immunotherapy; Lymphocytes; Lymphocytes T; Manufacturing; Mice; multidisciplinary; Neoplasms; Receptors; Receptors, Antigen, T-Cell - genetics; Receptors, Chimeric Antigen - genetics; Rejection; Science; Science (multidisciplinary); T-Lymphocytes; Tumors
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-023-37785-2

Manufacturing autologous chimeric antigen receptor (CAR) T cell therapeutics is complex, and many patients experience treatment delays or cannot be treated at all. Although current allogeneic CAR products have the potential to overcome manufacturing bottlenecks, they are subject to immune rejection and failure to persist in the host, and thus do not provide the same level of efficacy as their autologous counterparts. Here, we aimed to develop universal allogeneic CAR T cells that evade the immune system and produce a durable response. We generated human hypoimmune (HIP) T cells with disrupted B2M, CIITA , and TRAC genes using CRISPR-Cas9 editing. In addition, CD47 and anti-CD19 CAR were expressed using lentiviral transduction. These allogeneic HIP CD19 CAR T cells were compared to allogeneic CD19 CAR T cells that only expressed the anti-CD19 CAR (allo CAR T). In vitro assays for cancer killing and exhaustion revealed no differences between allo CAR T and HIP CAR T cells, confirming that the HIP edits did not negatively affect T cell performance. Clearance of CD19 + tumors by HIP CAR T cells in immunodeficient NSG mice was comparable to that of allo CAR T cells. In fully immunocompetent humanized mice, HIP CAR T cells significantly outperformed allo CAR T cells, showed improved persistence and expansion, and provided lasting cancer clearance. Furthermore, CD47-targeting safety strategies reliably and specifically eliminated HIP CAR T cells. These findings suggest that universal allogeneic HIP CAR T cell-based therapeutics might overcome the limitations associated with poor persistence of allogeneic CAR T cells and exert durable anti-tumor responses. The development of allogeneic chimaeric antigen receptor (CAR) T cells could overcome manufacturing bottlenecks for immunotherapy. However, immune rejection reduces the persistence and efficacy of these cells. Here, the authors generate allogeneic anti-CD19 CAR T cells that can evade the immune system and provide durable anti-tumour responses.