Combinatorial T cell engineering eliminates on-target off-tumor toxicity of CD229 CAR T cells while maintaining anti-tumor activity
bioRxiv (Cold Spring Harbor Laboratory)
Abstract
ABSTRACT T cells expressing chimeric antigen receptors have shown remarkable therapeutic activity against different types of cancer. However, their wider use has been hampered by the potential for life-threatening toxicities due to the unintended targeting of healthy cells expressing low levels of the targeted antigen. We have now developed an affinity-tuning approach for the generation of minimally modified, low-affinity antibody variants derived from existing high-affinity antibodies. Using this approach, we engineered low affinity variants of the fully human CD229-specific antibody 2D3. Parental 2D3 originally efficiently targeted multiple myeloma cells but also healthy T cells expressing low levels of CD229. We demonstrate that CAR T cells based on a low affinity variant of 2D3, engineered to also express CJUN to increase CAR T cell expansion, maintain the parental antibody’s anti-tumor activity but lack its targeting of healthy T cells in vitro and in vivo . In addition, we found that low affinity CD229 CAR T cells show reduced trogocytosis potentially augmenting CAR T cell persistence. The fast off-rate CAR produced using our affinity tuning approach eliminates a key liability of CD229 CAR T cells and paves the way for the effective and safe treatment of patients with multiple myeloma and other lymphoid malignancies. One sentence summary Rational T cell engineering yields low affinity CD229 CAR T cells overexpressing CJUN, which maintain the parental cells’ anti-tumor activity but eliminate killing of healthy T cells, increasing CAR T cell expansion, and decreasing trogocytosis.
Authors 13
-
University of Maryland, Baltimore · University of Utah · Huntsman Cancer Institute
Affiliation as printed
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD, United States
Department of Pharmaceutics & Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT, United States
Hematology and Hematologic Malignancies, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, United States
Department of Microbiology and Immunology, University of Maryland School of Medicine
Department of Pharmaceutics & Pharmaceutical Chemistry, University of Utah
Hematology and Hematologic Malignancies, Huntsman Cancer Institute, University of Utah
-
University of Maryland, Baltimore
Affiliation as printed
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD, United States
Department of Microbiology and Immunology, University of Maryland School of Medicine
-
Medical College of Wisconsin · University of Utah · Huntsman Cancer Institute
Affiliation as printed
Division of Hematology and Oncology, Medical College of Wisconsin, Milwaukee, USA
Hematology and Hematologic Malignancies, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, United States
Division of Hematology and Oncology, Medical College of Wisconsin
Hematology and Hematologic Malignancies, Huntsman Cancer Institute, University of Utah
-
University of Maryland, Baltimore · University of Maryland Marlene and Stewart Greenebaum Comprehensive Cancer Center
Affiliation as printed
Department of Medicine and Transplant/Cell Therapy Program, University of Maryland School of Medicine and Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, MD, United States
Department of Medicine and Transplant/Cell Therapy Program, University of Maryland School of Medicine and Marlene and Stewart Greenebaum Comprehensive Cancer Center
-
University of Utah · ARUP Laboratories (United States)
Affiliation as printed
Department of Pathology, University of Utah, Salt Lake City, UT, United States
Division of Experimental and Clinical Pathology, ARUP Laboratories, Salt Lake City, USA
Department of Pathology, University of Utah
Division of Experimental and Clinical Pathology
-
Universitätsklinikum Aachen · RWTH Aachen University
Affiliation as printed
Department of Oncology, Hematology, Hemostaseology, and Stem Cell Transplantation, University Hospital RWTH Aachen, Aachen, Germany
Department of Oncology, Hematology, Hemostaseology, and Stem Cell Transplantation, University Hospital RWTH Aachen
-
Affiliation as printed
Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, United States
Huntsman Cancer Institute, University of Utah
-
Affiliation as printed
Department of Pathology, University of Utah, Salt Lake City, UT, United States
Department of Pathology, University of Utah
-
University of Utah · Huntsman Cancer Institute
Affiliation as printed
Hematology and Hematologic Malignancies, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, United States
Hematology and Hematologic Malignancies, Huntsman Cancer Institute, University of Utah
-
Affiliation as printed
Department of Pathology, University of Utah, Salt Lake City, UT, United States
Department of Pathology, University of Utah
-
Affiliation as printed
Department of Pharmaceutics & Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT, United States
Department of Pharmaceutics & Pharmaceutical Chemistry, University of Utah
-
University of Maryland, Baltimore · University of Maryland Marlene and Stewart Greenebaum Comprehensive Cancer Center
Affiliation as printed
Department of Medicine and Transplant/Cell Therapy Program, University of Maryland School of Medicine and Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, MD, United States
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD, United States
Department of Medicine and Transplant/Cell Therapy Program, University of Maryland School of Medicine and Marlene and Stewart Greenebaum Comprehensive Cancer Center
Department of Microbiology and Immunology, University of Maryland School of Medicine
-
Tim Luetkens corresponding
University of Maryland, Baltimore · University of Utah · Huntsman Cancer Institute · University of Maryland Marlene and Stewart Greenebaum Comprehensive Cancer Center
Affiliation as printed
Department of Medicine and Transplant/Cell Therapy Program, University of Maryland School of Medicine and Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, MD, United States
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD, United States
Hematology and Hematologic Malignancies, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, United States
Department of Medicine and Transplant/Cell Therapy Program, University of Maryland School of Medicine and Marlene and Stewart Greenebaum Comprehensive Cancer Center
Department of Microbiology and Immunology, University of Maryland School of Medicine
Hematology and Hematologic Malignancies, Huntsman Cancer Institute, University of Utah
Cited by 1 stored of 1
1 result
No patents citing this paper on Lens.org (checked 2026-10-06).