Ebrahimi-Nik et al. identified a 10-amino acid neoepitope with very poor affinity for murine MHC in a murine fibrosarcoma. Vaccination of mice with mutated, but not unmutated, peptide induced CD8+ T cells that controlled challenge tumors. The tumor cell line was gene edited to create a revertant line, homozygous for the wild-type epitope. The revertant was then re-edited to create a third cell line, homozygous for the neoepitope. All three cell lines formed tumors with activated TILs at day 10; neoepitope+ tumors had fewer myeloid cells and more TILs. TILs in neoepitope+ tumors had higher expression of genes underlying cytotoxicity and TCR reactivity.
Contributed by Paula Hochman
ABSTRACT: High-affinity MHC I-peptide interactions are considered essential for immunogenicity. However, some neo-epitopes with low affinity for MHC I have been reported to elicit CD8 T cell dependent tumor rejection in immunization-challenge studies. Here we show in a mouse model that a neo-epitope that poorly binds to MHC I is able to enhance the immunogenicity of a tumor in the absence of immunization. Fibrosarcoma cells with a naturally occurring mutation are edited to their wild type counterpart; the mutation is then re-introduced in order to obtain a cell line that is genetically identical to the wild type except for the neo-epitope-encoding mutation. Upon transplantation into syngeneic mice, all three cell lines form tumors that are infiltrated with activated T cells. However, lymphocytes from the two tumors that harbor the mutation show significantly stronger transcriptional signatures of cytotoxicity and TCR engagement, and induce greater breadth of TCR reactivity than those of the wild type tumors. Structural modeling of the neo-epitope peptide/MHC I pairs suggests increased hydrophobicity of the neo-epitope surface, consistent with higher TCR reactivity. These results confirm the in vivo immunogenicity of low affinity or 'non-binding' epitopes that do not follow the canonical concept of MHC I-peptide recognition.
Author Info: (1) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. Broad Institute of MIT and Harv
Author Info: (1) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. Broad Institute of MIT and Harvard, 105 Broadway, Cambridge, MA, USA. (2) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. (3) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. (4) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. (5) Ludwig Institute for Cancer Research, University of Lausanne, Lausanne, Switzerland. Department of Oncology, Centre hospitalier universitaire vaudois (CHUV), Lausanne, Switzerland. (6) Ludwig Institute for Cancer Research, University of Lausanne, Lausanne, Switzerland. Department of Oncology, Centre hospitalier universitaire vaudois (CHUV), Lausanne, Switzerland. (7) Department of Medical Genetics, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan. Genome Medical Science Project, National Center for Global Health and Medicine, Chiba, Japan. (8) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. Arvinas, 5 science park, 395 Winchester Ave, New Haven, CT, USA. (9) Department of Chemistry and Biochemistry and Harper Cancer Research Institute, University of Notre Dame, Notre Dame, IN, USA. (10) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. (11) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. (12) Ludwig Institute for Cancer Research, University of Lausanne, Lausanne, Switzerland. Department of Oncology, Centre hospitalier universitaire vaudois (CHUV), Lausanne, Switzerland. (13) Department of Chemistry and Biochemistry and Harper Cancer Research Institute, University of Notre Dame, Notre Dame, IN, USA. (14) Department of Computer Sciences, University of Connecticut School of Engineering, Storrs, CT, USA. (15) Ludwig Institute for Cancer Research, University of Lausanne, Lausanne, Switzerland. Department of Oncology, Centre hospitalier universitaire vaudois (CHUV), Lausanne, Switzerland. (16) Department of Immunology and Carole and Ray Neag Comprehensive Cancer Center, University of Connecticut School of Medicine, Farmington, CT, USA. Srivastava@uchc.edu.
