Inducible de novo expression of neoantigens in tumor cells and mice

• Published in: Nature biotechnology 2021-01, Vol.39 (1), p.64-73
• Main Authors: Damo, Martina, Fitzgerald, Brittany, Lu, Yisi, Nader, Mursal, William, Ivana, Cheung, Julie F., Connolly, Kelli A., Foster, Gena G., Akama-Garren, Elliot, Lee, Da-Yae, Chang, Greg P., Gocheva, Vasilena, Schmidt, Leah M., Boileve, Alice, Wilson, Josephine H., Cui, Can, Monroy, Isabel, Gokare, Prashanth, Cabeceiras, Peter, Jacks, Tyler, Joshi, Nikhil S.
• Format: Article
• Language: English
• Subjects: 631/1647/767/1972; 631/250/2152/569/2494; 631/250/2152/569/2495; 631/61/17/1511; Agriculture; Analysis; Animals; Antigens; Antigens, Neoplasm - genetics; Antigens, Neoplasm - metabolism; Autoimmune diseases; Bioinformatics; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Cancer; CD4 antigen; CD4-Positive T-Lymphocytes - chemistry; CD4-Positive T-Lymphocytes - metabolism; CD8 antigen; CD8-Positive T-Lymphocytes - chemistry; CD8-Positive T-Lymphocytes - metabolism; Cell Engineering - methods; Deoxyribonucleic acid; DNA; Female; Gene expression; Genetic aspects; Health aspects; Humans; Immunological tolerance; Life Sciences; Lymphocytes; Lymphocytes T; Mice; Neoantigens; Oncology, Experimental; Organ Specificity - genetics; Recombination; Ribonucleic acid; RNA; RNA Splicing - genetics; Splicing; T cells; Transplantation; Tumor antigens; Tumor cell lines; Tumor cells; Tumor Cells, Cultured; Tumors
• ISSN: 1087-0156; 1546-1696; 1546-1696
• DOI: 10.1038/s41587-020-0613-1

Inducible expression of neoantigens in mice would enable the study of endogenous antigen-specific naïve T cell responses in disease and infection, but has been difficult to generate because leaky antigen expression in the thymus results in central T cell tolerance. Here we develop inversion-induced joined neoantigen (NINJA), using RNA splicing, DNA recombination and three levels of regulation to prevent leakiness and allow tight control over neoantigen expression. We apply NINJA to create tumor cell lines with inducible neoantigen expression, which could be used to study antitumor immunity. We also show that the genetic regulation in NINJA mice bypasses central and peripheral tolerance mechanisms and allows for robust endogenous CD8 and CD4 T cell responses on neoantigen induction in peripheral tissues. NINJA will enable studies of how T cells respond to defined neoantigens in the context of peripheral tolerance, transplantation, autoimmune diseases and cancer. A system to express neoantigens in tumor cells and mice circumvents central T cell tolerance.