Background: To induce robust cellular immunity against non-small cell lung cancer (NSCLC), the leading cause of cancer-related death in the US, we have developed an innovative cell-based therapeutic platform technology to generate polyclonal, neoantigen-reactive T cells in vitro. Methods: Surgically resected NSCLC tissue and peripheral matched blood were collected from 17 patients under an IRB-approved protocol. From 3 of the patient-derived NSCLC specimens, methods were tested to enhance immunogenicity and stimulate neoantigen-specific immune responses. We then use a novel, human vascularized microtumor (VMT) platform to assess the sensitivity of tumor cells to T-cell mediated attack at the individual patient level. Results: The therapeutic platform induced a significant increase in T cell proliferation, interferon-gamma secretion, and blast phenotype of lymphocyte populations compared to T cells primed by standard methods. Interestingly, for one patient, immunosuppression of the T cell response with concomitant increase in CD96, an immune checkpoint receptor implicated in lung cancer progression, was observed in the tumor. Importantly, using sequencing data from one patient, an analytical and machine learning pipeline detected putative neoantigens using our therapeutic platform technology and not with standard methods. Conclusions: The therapeutic platform can be used to directly generate cell-based immunotherapies and to facilitate neoantigen discovery and development of targeted cancer immunotherapies. Our goal is to advance novel approaches to effectively mobilize anti-tumor immunity and overcome treatment resistance in NSCLC.