GPRC5D is a target for the immunotherapy of multiple myeloma with rationally designed CAR T cells

Eric L. Smith, Kim Harrington, Mette Staehr, Reed Masakayan, Jon Jones, Thomas J. Long, Khong Y. Ng, Majid Ghoddusi, Terence J. Purdon, Xiuyan Wang, Trevor Do, Minh Thu Pham, Jessica M. Brown, Carlos Fernandez De Larrea, Eric Olson, Elizabeth Peguero, Pei Wang, Hong Liu, Yiyang Xu, Sarah C. Garrett-Thomson & 6 others Steven C. Almo, Hans Guido Wendel, Isabelle Riviere, Cheng Liu, Blythe Sather, Renier J. Brentjens

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Article numbereaau7746
JournalScience Translational Medicine
Volume11
Issue number485
DOIs
StatePublished - Jan 1 2019

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B-Cell Maturation Antigen
G-Protein-Coupled Receptors
T-Cell Antigen Receptor
Multiple Myeloma
Immunotherapy
Antigen Receptors
Bone Marrow
Cell- and Tissue-Based Therapy
Antigens
Myeloma Proteins
Single-Chain Antibodies
Hair Follicle
Alopecia
Heterografts
Bacteriophages
Libraries
Fluorescent Antibody Technique
B-Lymphocytes
Clone Cells
Cytokines

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Smith, E. L., Harrington, K., Staehr, M., Masakayan, R., Jones, J., Long, T. J., ... Brentjens, R. J. (2019). GPRC5D is a target for the immunotherapy of multiple myeloma with rationally designed CAR T cells. Science Translational Medicine, 11(485), [eaau7746]. https://doi.org/10.1126/scitranslmed.aau7746

GPRC5D is a target for the immunotherapy of multiple myeloma with rationally designed CAR T cells. / Smith, Eric L.; Harrington, Kim; Staehr, Mette; Masakayan, Reed; Jones, Jon; Long, Thomas J.; Ng, Khong Y.; Ghoddusi, Majid; Purdon, Terence J.; Wang, Xiuyan; Do, Trevor; Pham, Minh Thu; Brown, Jessica M.; De Larrea, Carlos Fernandez; Olson, Eric; Peguero, Elizabeth; Wang, Pei; Liu, Hong; Xu, Yiyang; Garrett-Thomson, Sarah C.; Almo, Steven C.; Wendel, Hans Guido; Riviere, Isabelle; Liu, Cheng; Sather, Blythe; Brentjens, Renier J.

In: Science Translational Medicine, Vol. 11, No. 485, eaau7746, 01.01.2019.

Research output: Contribution to journalArticle

Smith, EL, Harrington, K, Staehr, M, Masakayan, R, Jones, J, Long, TJ, Ng, KY, Ghoddusi, M, Purdon, TJ, Wang, X, Do, T, Pham, MT, Brown, JM, De Larrea, CF, Olson, E, Peguero, E, Wang, P, Liu, H, Xu, Y, Garrett-Thomson, SC, Almo, SC, Wendel, HG, Riviere, I, Liu, C, Sather, B & Brentjens, RJ 2019, 'GPRC5D is a target for the immunotherapy of multiple myeloma with rationally designed CAR T cells', Science Translational Medicine, vol. 11, no. 485, eaau7746. https://doi.org/10.1126/scitranslmed.aau7746
Smith, Eric L. ; Harrington, Kim ; Staehr, Mette ; Masakayan, Reed ; Jones, Jon ; Long, Thomas J. ; Ng, Khong Y. ; Ghoddusi, Majid ; Purdon, Terence J. ; Wang, Xiuyan ; Do, Trevor ; Pham, Minh Thu ; Brown, Jessica M. ; De Larrea, Carlos Fernandez ; Olson, Eric ; Peguero, Elizabeth ; Wang, Pei ; Liu, Hong ; Xu, Yiyang ; Garrett-Thomson, Sarah C. ; Almo, Steven C. ; Wendel, Hans Guido ; Riviere, Isabelle ; Liu, Cheng ; Sather, Blythe ; Brentjens, Renier J. / GPRC5D is a target for the immunotherapy of multiple myeloma with rationally designed CAR T cells. In: Science Translational Medicine. 2019 ; Vol. 11, No. 485.
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abstract = "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.",
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AU - Smith, Eric L.

AU - Harrington, Kim

AU - Staehr, Mette

AU - Masakayan, Reed

AU - Jones, Jon

AU - Long, Thomas J.

AU - Ng, Khong Y.

AU - Ghoddusi, Majid

AU - Purdon, Terence J.

AU - Wang, Xiuyan

AU - Do, Trevor

AU - Pham, Minh Thu

AU - Brown, Jessica M.

AU - De Larrea, Carlos Fernandez

AU - Olson, Eric

AU - Peguero, Elizabeth

AU - Wang, Pei

AU - Liu, Hong

AU - Xu, Yiyang

AU - Garrett-Thomson, Sarah C.

AU - Almo, Steven C.

AU - Wendel, Hans Guido

AU - Riviere, Isabelle

AU - Liu, Cheng

AU - Sather, Blythe

AU - Brentjens, Renier J.

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