Finding that CCR7 was downregulated during CAR T manufacturing, Zschummel et al. generated CAR T cells with CCR7 overexpression (CAR.CCR7). Compared to CAR-T, CAR.CCR7-T had increased CCL19-mediated migration in vitro, and demonstrated LN accumulation and lymphoma depletion in a syngeneic mouse model. CAR.CCR7-T had superior cytotoxicity in vitro compared to standard CAR-T, especially at low E:T, but not due to CAR or Th1 cytokine expression, or the ligand CCL19. CAR.CCR7-T cells were larger, had increased cytoskeletal regulatory gene expression, recruited CCR7 to the immune synapse, and showed early ZAP70 phosphorylation and degranulation.
Contributed by Alex Najibi
ABSTRACT: Anti-CD19 chimeric antigen receptor (CAR) therapy demonstrated remarkable efficacy against hematological malignancies. However, B cell malignancies with lymph node (LN) involvement frequently remain resistant. Here, we show that CAR T cells downregulated the chemokine receptor CCR7, crucial for nodal homing, during manufacturing. Consequently, in vitro migration toward the respective chemokines and in vivo migration to LNs was severely impaired. To improve nodal CAR T-cell trafficking, we engineered anti-CXCR5 CAR T cells, targeting mature lymphoma, with stable CCR7 expression (CAR.CCR7). CCR7 engineering of human and mouse CAR T cells restored migratory capacity and LN homing. Additionally, we observed enhanced CAR-mediated killing in CCR7-engineered anti-CXCR5 and anti-CD19-CARs alike, a process that was independent of increased cytokine secretion. Mechanistically, CCR7 overexpression was associated with an altered expression of genes involved in cytoskeletal rearrangement and faster killing kinetics. CCR7 accumulated in mature CAR synapses, supporting the costimulatory role of CCR7 within immunological synapses. Therapeutically, improved LN-recruitment and enhanced killing of CAR.CCR7 T cells improved lymphoma eradication in mice.
Author Info: (1) Massachusetts General Hospital Charlestown, Massachusetts United States. ROR: https://ror.org/002pd6e78 (2) Max Delbrck Center for Molecular Medicine Berlin, Berlin Germany. R
Author Info: (1) Massachusetts General Hospital Charlestown, Massachusetts United States. ROR: https://ror.org/002pd6e78 (2) Max Delbrck Center for Molecular Medicine Berlin, Berlin Germany. ROR: https://ror.org/04p5ggc03 (3) Max Delbrck Center for Molecular Medicine Berlin Germany. ROR: https://ror.org/04p5ggc03 (4) Max Delbrck Center Berlin, Berlin Germany. (5) Max Delbrck Center for Molecular Medicine Berlin Germany. ROR: https://ror.org/04p5ggc03 (6) Max Delbrck Center for Molecular Medicine Berlin Germany. ROR: https://ror.org/04p5ggc03 (7) Max Delbrck Center for Molecular Medicine Berlin Germany. (8) Max Delbrck Center for Molecular Medicine Berlin Germany. (9) Max Delbrck Center for Molecular Medicine Berlin Germany. (10) Max Delbrck Center for Molecular Medicine Berlin Germany. (11) Max Delbrck Center for Molecular Medicine Berlin Germany. ROR: https://ror.org/04p5ggc03 (12) Max Delbrck Center for Molecular Medicine Berlin Germany. ROR: https://ror.org/04p5ggc03
