CD28 Co-Stimulus Achieves Superior CAR T Cell Effector Function against Solid Tumors Than 4-1BB Co-Stimulus

Cancers, vol. 13, pp. 1050

Abstract

Spacer or co-stimulatory components in chimeric antigen receptor (CAR) design influence CAR T cell effector function. Few preclinical mouse models optimally support CAR candidate pre-selection for clinical development. Here we use a model in which murine CAR T cells can be exploited with human tumor xenografts. This mouse-in-mouse approach avoids limitations caused by species-specific factors crucial for CAR T cell survival, trafficking and function. We compared trafficking, expansion and tumor control for T cells expressing different CAR construct designs targeting two antigens (L1CAM or HER2), structurally identical except for spacer (long or short) or co-stimulatory (4-1BB or CD28) domains to be evaluated. Using monoclonal, murine-derived L1CAM-specific CAR T cells in Rag-/- mice harboring established xenografted tumors from a human neuroblastoma cell line revealed a clear superiority in CAR T cell trafficking using CD28 co-stimulation. L1CAM-targeting short spacer-CD28/ζ CAR T cells expanded the most at the tumor site and induced initial tumor regression. Treating patient-derived neuroblastoma xenografts with human L1CAM-targeting CAR T cells confirmed the superiority of CD28 co-stimulus. CD28 superiority was also demonstrated with HER2-specific CAR T cells (targeting ovarian carcinoma xenografts). Our findings encourage incorporating CD28 signaling into CAR design for adoptive T cell treatment of solid tumors.

Authors 15

1. Ana Textor Aachen Department of Hematology, Leiden University Medical Center

   Max Delbrück Center · Leiden University Medical Center · Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Hematology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands

   Institute of Immunology, Charité Campus Buch, 13092 Berlin, Germany

   Max-Delbrück-Center for Molecular Medicine (MDC), Robert-Rössle-Straße 10, 13092 Berlin, Germany

2. Laura Grunewald

   Humboldt-Universität zu Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

3. Kathleen Anders

   Humboldt-Universität zu Berlin · Deutsches Konsortium für Translationale Krebsforschung · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

   German Cancer Consortium (DKTK), Partner Site Berlin, CCCC (Campus Mitte), 10178 Berlin, Germany

4. Anika Klaus

   Humboldt-Universität zu Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

5. Silke Schwiebert

   Humboldt-Universität zu Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

6. Annika Winkler

   Humboldt-Universität zu Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

7. Maria Stecklum
   Affiliation as printed

   Experimental Pharmacology & Oncology Berlin Buch, 13125 Berlin, Germany

8. Jana Rolff
   Affiliation as printed

   Experimental Pharmacology & Oncology Berlin Buch, 13125 Berlin, Germany

9. Anton George Henssen

   Humboldt-Universität zu Berlin · Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Berlin Institute of Health (BIH), 10178 Berlin, Germany

   Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

   Experimental and Clinical Research Center, Lindenberger Weg 80, 13125 Berlin, Germany

10. Uta E. Höpken

    Max Delbrück Center

    Affiliation as printed

    Max-Delbrück-Center for Molecular Medicine (MDC), Robert-Rössle-Straße 10, 13092 Berlin, Germany

11. Angelika Eggert

    German Cancer Research Center · Humboldt-Universität zu Berlin · Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Deutsches Konsortium für Translationale Krebsforschung · Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Berlin Institute of Health (BIH), 10178 Berlin, Germany

    Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

    German Cancer Consortium (DKTK), Partner Site Berlin, CCCC (Campus Mitte), 10178 Berlin, Germany

    German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany

12. Johannes Hubertus Schulte

    German Cancer Research Center · Humboldt-Universität zu Berlin · Deutsches Konsortium für Translationale Krebsforschung · Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

    German Cancer Consortium (DKTK), Partner Site Berlin, CCCC (Campus Mitte), 10178 Berlin, Germany

    German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany

13. Michael C. Jensen

    University of Washington · Fred Hutch Cancer Center · Seattle Children's Research Institute

    Affiliation as printed

    Department of Bioengineering, University of Washington, Seattle, WA 98195, USA

    Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA

    Seattle Children’s Therapeutics, Seattle Children’s Research Institute, Seattle, WA 98101, USA

    Seattle Children's Therapeutics, Seattle Children's Research Institute, Seattle, WA 98101, USA

14. Thomas J. F. Blankenstein

    Max Delbrück Center · Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Institute of Immunology, Charité Campus Buch, 13092 Berlin, Germany

    Max-Delbrück-Center for Molecular Medicine (MDC), Robert-Rössle-Straße 10, 13092 Berlin, Germany

15. Annette Künkele corresponding

    German Cancer Research Center · Humboldt-Universität zu Berlin · Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Deutsches Konsortium für Translationale Krebsforschung · Charité - Universitätsmedizin Berlin

    Affiliation as printed

    Berlin Institute of Health (BIH), 10178 Berlin, Germany

    Department of Pediatric Oncology and Hematology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany

    German Cancer Consortium (DKTK), Partner Site Berlin, CCCC (Campus Mitte), 10178 Berlin, Germany

    German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany