A functional identification platform reveals frequent, spontaneous neoantigen-specific T cell responses in patients with cancer
Aaron M. Miller, Zeynep Koşaloğlu, Luise Westernberg, Leslie Montero, Milad Bahmanof, Angela Frentzen, Manasa Lanka, Ashmitaa Logandha Ramamoorthy Premlal, Grégory Seumois, Jason Greenbaum, Spencer E. Brightman, Karla Soria Zavala, Rukman R. Thota, Martin S. Naradikian, Samir Makani, Scott M. Lippman, Alessandro Sette, Ezra E.W. Cohen, Bjoern Peters, Stephen P. Schoenberger
Abstract
The clinical impact of tumor-specific neoantigens as both immunotherapeutic targets and biomarkers has been impeded by the lack of efficient methods for their identification and validation from routine samples. We have developed a platform that combines bioinformatic analysis of tumor exomes and transcriptional data with functional testing of autologous peripheral blood mononuclear cells (PBMCs) to simultaneously identify and validate neoantigens recognized by naturally primed CD4 + and CD8 + T cell responses across a range of tumor types and mutational burdens. The method features a human leukocyte antigen (HLA)–agnostic bioinformatic algorithm that prioritizes mutations recognized by patient PBMCs at a greater than 40% positive predictive value followed by a short-term in vitro functional assay, which allows interrogation of 50 to 75 expressed mutations from a single 50-ml blood sample. Neoantigens validated by this method include both driver and passenger mutations, and this method identified neoantigens that would not have been otherwise detected using an in silico prediction approach. These findings reveal an efficient approach to systematically validate clinically actionable neoantigens and the T cell receptors that recognize them and demonstrate that patients across a variety of human cancers have a diverse repertoire of neoantigen-specific T cells.