Abstract LB-150: Neoantigens that fail to elicit measurable T cell responses following peptide immunization can control tumor growth when delivered using a Listeria-based immunotherapy platform
Authors: Coder Brandon , Phee Hyewon , Mottershead Cristina , Kelkar Dipti , Filippova Elena , Ju Xiaoming , Lugt Bryan Vander , Pryshchep Olga , Lesch Justin , Liu Xian , DeVoss Jason , Cooke Keegan , Liu Claret , Zhan Jinghui , Mitchell Petia , Ramos Kim , Villarreal Daniel O. , Johnston Jim , Petit Robert , Princiotta Michael 
Published: August 2018

Abstract
Introduction: Recent advances in the field of cancer immunotherapy have identified CD8+ T cell responses against tumor-specific neoantigens as a key driver of tumor regression and prolonged survival. ADXS-NEO is a personalized Listeria monocytogenes (Lm)-based immunotherapy designed to generate immune responses against mutation-derived tumor-specific neoantigens. Advaxis' Lm-based immunotherapies consist of live highly-attenuated bacterial vectors that are bioengineered to secrete a fusion protein consisting of a truncated non-hemolytic fragment of listeriolysin O, which has adjuvant properties, and tumor-specific neoantigens that harbor nonsynonymous point mutations (NSMs). The objective of this study is to demonstrate the feasibility of using the ADXS-NEO platform to target tumor-specific point mutations to generate neoantigen-specific T cells and control tumor growth.
Results: Whole-exome sequencing of the MC38 mouse tumor cell line identified 2870 unique NSMs. Among these, the IC50 of 138 NSMs were predicted to be less than 500 nM by the netMHCcons algorithm. We evaluated the immunogenicity of 37 NSMs, and found that 12 immunogenic NSMs elicited a CD8+ T cell response following peptide immunization. Moreover, we identified 10 additional immunogenic NSMs in MC38-bearing mice treated with a check point inhibitor. Altogether, we identified 22 immunogenic and 23 non-immunogenic NSMs. Two ADXS-NEO vectors were constructed, Lm-19 & Lm-20, targeting 19 non-immunogenic and 20 immunogenic NSMs respectively. The ability of Lm-19 and Lm-20 to control MC38 tumor growth was evaluated in C57BL/6J mice. We found that both Lm-19 & Lm-20 led to an accumulation of neoantigen-specific CD8+ TILs and significantly slowed tumor growth. Moreover, both Lm-19 and Lm-20 decreased the frequency and absolute number of intratumoral Tregs, TAMs, and MDSCs and increased the frequency and absolute number of effector CD8+ T cells. Interestingly, expression of PD-L1 was decreased in TAMs and MDSCs and the frequency and total number of granzyme A+ CD8+ effector T cells was increased. Furthermore, the proportion of phenotypically exhausted PD-1hiLAG3+ TILs was decreased. Together, these data suggest the tumor microenvironment in mice receiving Lm-19 and Lm-20 becomes more cytotoxic and less suppressive.
Conclusion: ADXS-NEO is a potent immunotherapy capable of driving immune responses against tumor-specific mutations and leading to tumor control. The effectiveness of the Lm platform is demonstrated by the generation of neoantigen-specific T cells to peptide sequences that were identified as “non-immunogenic” using a conventional peptide-adjuvant immunization. This study is a clear demonstration that T cell mediated anti-tumor responses can be generated by targeting tumor-derived NSMs with the ADXS-NEO Listeria monocytogenes vector.
Citation Format: Brandon Coder, Hyewon Phee, Cristina Mottershead, Dipti Kelkar, Elena Filippova, Xiaoming Ju, Bryan Vander Lugt, Olga Pryshchep, Justin Lesch, Xian Liu, Jason DeVoss, Keegan Cooke, Claret Liu, Jinghui Zhan, Petia Mitchell, Kim Ramos, Daniel O. Villarreal, Jim Johnston, Robert Petit, Michael Princiotta. Neoantigens that fail to elicit measurable T cell responses following peptide immunization can control tumor growth when delivered using a Listeria-based immunotherapy platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-150.