Having demonstrated that GPRC5D, normally found only in the immune-privileged tissue of hair follicles, is expressed on multiple myeloma (MM) cells, independent of BCMA (B cell maturation antigen) expression, Smith et al. utilized a human B cell-derived scFv phage display library and Jurkat Nur77 reporter T cells in vitro and MM xenografts in vivo to identify and optimize a GPRC5D-specific CAR construct. Anti-GPRC5D CAR T cells exhibited cytotoxic activity similar to anti-BCMA CAR T cells, eradicated MM even in case of BCMA antigen loss relapse, and promoted long-term survival in mice. No fur loss or other toxicities were observed in mice or monkeys.
Early clinical results of chimeric antigen receptor (CAR) T cell therapy targeting B cell maturation antigen (BCMA) for multiple myeloma (MM) appear promising, but relapses associated with residual low-to-negative BCMA-expressing MM cells have been reported, necessitating identification of additional targets. The orphan G protein-coupled receptor, class C group 5 member D (GPRC5D), normally expressed only in the hair follicle, was previously identified as expressed by mRNA in marrow aspirates from patients with MM, but confirmation of protein expression remained elusive. Using quantitative immunofluorescence, we determined that GPRC5D protein is expressed on CD138(+) MM cells from primary marrow samples with a distribution that was similar to, but independent of, BCMA. Panning a human B cell-derived phage display library identified seven GPRC5D-specific single-chain variable fragments (scFvs). Incorporation of these into multiple CAR formats yielded 42 different constructs, which were screened for antigen-specific and antigen-independent (tonic) signaling using a Nur77-based reporter system. Nur77 reporter screen results were confirmed in vivo using a marrow-tropic MM xenograft in mice. CAR T cells incorporating GPRC5D-targeted scFv clone 109 eradicated MM and enabled long-term survival, including in a BCMA antigen escape model. GPRC5D(109) is specific for GPRC5D and resulted in MM cell line and primary MM cytotoxicity, cytokine release, and in vivo activity comparable to anti-BCMA CAR T cells. Murine and cynomolgus cross-reactive CAR T cells did not cause alopecia or other signs of GPRC5D-mediated toxicity in these species. Thus, GPRC5D(109) CAR T cell therapy shows potential for the treatment of advanced MM irrespective of previous BCMA-targeted therapy.
Author Info: (1) Cellular Therapeutics Center, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. Myeloma Service, Department of Medicine, Memorial Sloan K
Author Info: (1) Cellular Therapeutics Center, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. Myeloma Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. (2) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (3) Cellular Therapeutics Center, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. (4) Cellular Therapeutics Center, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. (5) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (6) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (7) Sloan Kettering Institute, New York, NY 10065, USA. (8) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (9) Cellular Therapeutics Center, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. (10) Cell Therapy and Cell Engineering Facility, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. (11) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (12) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (13) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (14) Cellular Therapeutics Center, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. Amyloidosis and Myeloma Unit, Department of Hematology, Hospital Clinic, August Pi i Sunyer Biomedical Research Institute, University of Barcelona, 08036 Barcelona, Spain. (15) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (16) Sloan Kettering Institute, New York, NY 10065, USA. (17) Eureka Therapeutics, Emeryville, CA 94608, USA. (18) Eureka Therapeutics, Emeryville, CA 94608, USA. (19) Eureka Therapeutics, Emeryville, CA 94608, USA. (20) Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461, USA. (21) Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461, USA. (22) Sloan Kettering Institute, New York, NY 10065, USA. (23) Cell Therapy and Cell Engineering Facility, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. (24) Eureka Therapeutics, Emeryville, CA 94608, USA. (25) Juno Therapeutics, A Celgene Company, Seattle, WA 98109, USA. (26) Cellular Therapeutics Center, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. brentjer@mskcc.org. Leukemia Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA.
