Abstract 4388: The role of neoantigens in immunotherapy of cutaneous melanoma

Melanoma is increasing in incidence by up to 3% annually, and metastatic melanoma, which is resistant to most conventional therapies, has a poor prognosis with a 5-year survival rate of less than 20%. Immunotherapy is able to yield durable response in melanoma; however, the response rate is limited (20-30%). It has been shown that tumor immunogenicity is highly correlated to the response to immunotherapy. The mutational load and the characteristics of the neoantigens displayed by the tumor cells are theorized to play an important role in the immunogenicity of the tumor. We used genetically engineered preclinical mouse models to study the role of these neoantigens in melanoma response to immune therapy. Three syngeneic C57BL/6 mouse melanoma models were developed: UV induced melanoma in albino BRAF(V600E);PTEN-/- mice (1), DMBA induced melanoma in pigmented HGF-tg;CDK4(R24C) mice (2), and UV induced melanoma in pigmented HGF-tg mice. These three models were found to have varying degrees of tumor immunogenicity based on vaccination studies. The UV induced BRAF(V600E);PTEN-/- model displayed poor immunogenicity, while the DMBA induced HGF-tg;CDK4(R24C) model exhibited moderate immunogenicity and the UV induced HGF-tg model showed the highest immunogenicity. Exome sequencing results showed that high immunogenicity is associated with mutations of genes in DNA damage responses and frameshift mutations. To test the role of neoantigens, RNA from each of the melanoma models was sequenced and analyzed for mutational and expression profiles. We will be comparing the response of these three mouse models to correlate immunogenicity with response to anti-CTLA-4 therapy. We plan to analyze the pathways in which mutated genes were enriched, allowing the identification of “druggable” targets to enhance therapeutic efficacy of immune checkpoint inhibitors. CRISPR/Cas9-based screening approaches will be used to identify specific neoantigens able to influence response to immune therapy. In conclusion, our preclinical mouse models show that tumor immunogenicity may be correlated to the carcinogenic mechanisms, and RNA sequence has provided further insight into the role of mutational load and neo-epitopes on immune-based therapeutic response. (1) Provided by Marcus Bosenberg (Yale School of Medicine, New Heaven, CT), Martin McMahon (Huntsman Cancer Institute, Salt Lake City, UT). (2) Provided by Thomas Tueting (University Hospital Bonn, Bonn, Germany). Citation Format: Renee M.