Sarah Oelsner1, Miriam E Friede1, Congcong Zhang1, Juliane Wagner2, Susanne Badura3, Peter Bader4, Evelyn Ullrich2, Oliver G Ottmann5, Hans Klingemann6, Torsten Tonn7, Winfried S Wels8. 1. Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany. 2. Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University Frankfurt, Frankfurt, Germany; LOEWE Center for Cell and Gene Therapy, Goethe University Frankfurt, Frankfurt, Germany. 3. Department of Medicine, Hematology and Oncology, Goethe University Frankfurt, Frankfurt, Germany. 4. Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University Frankfurt, Frankfurt, Germany. 5. Department of Haematology, Division of Cancer and Genetics, Cardiff University School of Medicine, Cardiff, United Kingdom. 6. NantKwest, Inc., Culver City, California, USA. 7. Institute for Transfusion Medicine, German Red Cross Blood Donation Service North-East, Dresden and Transfusion Medicine, Medical Faculty Carl Gustav Carus, TU Dresden, Germany. 8. Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany; German Cancer Consortium (DKTK), partner site Frankfurt/Mainz, Germany. Electronic address: wels@gsh.uni-frankfurt.de.
Abstract
BACKGROUND AIMS: Natural killer (NK) cells can rapidly respond to transformed and stressed cells and represent an important effector cell type for adoptive immunotherapy. In addition to donor-derived primary NK cells, continuously expanding cytotoxic cell lines such as NK-92 are being developed for clinical applications. METHODS: To enhance their therapeutic utility for the treatment of B-cell malignancies, we engineered NK-92 cells by lentiviral gene transfer to express chimeric antigen receptors (CARs) that target CD19 and contain human CD3ζ (CAR 63.z), composite CD28-CD3ζ or CD137-CD3ζ signaling domains (CARs 63.28.z and 63.137.z). RESULTS: Exposure of CD19-positive targets to CAR NK-92 cells resulted in formation of conjugates between NK and cancer cells, NK-cell degranulation and selective cytotoxicity toward established B-cell leukemia and lymphoma cells. Likewise, the CAR NK cells displayed targeted cell killing of primary pre-B-ALL blasts that were resistant to parental NK-92. Although all three CAR NK-92 cell variants were functionally active, NK-92/63.137.z cells were less effective than NK-92/63.z and NK-92/63.28.z in cell killing and cytokine production, pointing to differential effects of the costimulatory CD28 and CD137 domains. In a Raji B-cell lymphoma model in NOD-SCID IL2R γnull mice, treatment with NK-92/63.z cells, but not parental NK-92 cells, inhibited disease progression, indicating that selective cytotoxicity was retained in vivo. CONCLUSIONS: Our data demonstrate that it is feasible to generate CAR-engineered NK-92 cells with potent and selective antitumor activity. These cells may become clinically useful as a continuously expandable off-the-shelf cell therapeutic agent.
BACKGROUND AIMS: Natural killer (NK) cells can rapidly respond to transformed and stressed cells and represent an important effector cell type for adoptive immunotherapy. In addition to donor-derived primary NK cells, continuously expanding cytotoxic cell lines such as NK-92 are being developed for clinical applications. METHODS: To enhance their therapeutic utility for the treatment of B-cell malignancies, we engineered NK-92 cells by lentiviral gene transfer to express chimeric antigen receptors (CARs) that target CD19 and contain human CD3ζ (CAR 63.z), composite CD28-CD3ζ or CD137-CD3ζ signaling domains (CARs 63.28.z and 63.137.z). RESULTS: Exposure of CD19-positive targets to CAR NK-92 cells resulted in formation of conjugates between NK and cancer cells, NK-cell degranulation and selective cytotoxicity toward established B-cell leukemia and lymphoma cells. Likewise, the CAR NK cells displayed targeted cell killing of primary pre-B-ALL blasts that were resistant to parental NK-92. Although all three CAR NK-92 cell variants were functionally active, NK-92/63.137.z cells were less effective than NK-92/63.z and NK-92/63.28.z in cell killing and cytokine production, pointing to differential effects of the costimulatory CD28 and CD137 domains. In a Raji B-cell lymphoma model in NOD-SCID IL2R γnull mice, treatment with NK-92/63.z cells, but not parental NK-92 cells, inhibited disease progression, indicating that selective cytotoxicity was retained in vivo. CONCLUSIONS: Our data demonstrate that it is feasible to generate CAR-engineered NK-92 cells with potent and selective antitumor activity. These cells may become clinically useful as a continuously expandable off-the-shelf cell therapeutic agent.
Authors: Sunil S Raikar; Lauren C Fleischer; Robert Moot; Andrew Fedanov; Na Yoon Paik; Kristopher A Knight; Christopher B Doering; H Trent Spencer Journal: Oncoimmunology Date: 2017-12-26 Impact factor: 8.110
Authors: Rasa Islam; Aleta Pupovac; Vera Evtimov; Nicholas Boyd; Runzhe Shu; Richard Boyd; Alan Trounson Journal: Cells Date: 2021-04-29 Impact factor: 6.600