Abstract B118: Development of a novel prostatic acid phosphatase-derived vaccine for the treatment of advanced prostate cancer
Authors: Vu Pauline Le , Vadakekolathu Jayakumar , Christensen Dennis , Durrant Lindy , Pockley Graham , McArdle Stephanie E.B. 
Published: February 2019

Abstract
Our aim is to develop a new and more effective prostatic acid phosphatase (PAP) peptide-based vaccine for the treatment of advanced prostate cancer (PCa). We have previously shown that a 15mer PAP peptide-derived vaccine, when administered as a DNA vaccine, could induce PAP-specific T-cell responses and reduce the growth of implanted murine TRAMP C1-derived tumors in a syngeneic heterotopic murine model. We have subsequently developed an elongated (42mer) PAP-derived peptide in which Alanine at position 115 was replaced by Leucine.Herein, the capacity of the wild-type and mutated PAP-42mer vaccines to induce PAP-specific immune responses were compared in two pre-clinical murine models. The murine PAP-42mer sequence was evaluated in C57Bl/6 mice and the human PAP-42mer sequence in HHDII/DR1 transgenic mice. Following the identification of the most promising PAP-42mer sequence, the capacities of different delivery systems (CpG adjuvant, CAF09 adjuvant and ImmunoBody® DNA vaccine) to generate PAP-specific immunity were compared. For these studies, the responsiveness of splenocytes isolated from immunized mice following in vitro stimulation with MHC class I or class II wild-type vaccine-derived peptides (8/9 or 15 amino-acid long) was assessed using an IFNγ ELISpot assay and by immunophenotyping of splenic T-cells using flow cytometry. The ability of splenocytes to kill relevant targeT-cells after a short in vitro stimulation was assessed using a 51chromium release assay. B16 cells that had been knocked out for beta-2m gene and transfected to express chimeric HLA-A2 (HHDII), HLA-DR1 and human PAP implanted into HHDII/DR1 mice was used as a proof-of-concept model to assess the antitumor efficacy of the hPAP42mer mutated vaccine with CAF09 adjuvant in a prophylactic setting.The mutated murine and human PAP42mer-based vaccines induced a higher number of IFNγ-releasing splenocytes in response to in vitro stimulation with class I or class II vaccine-derived peptides and generated cells having a higher functional avidity. CAF09 and ImmunoBody® were superior to CpG in inducing PAP-specific immune responses, with CAF09 eliciting strong immune responses in both models, and ImmunoBody® eliciting potent immune responses in C57Bl/6 mice. Indeed, a higher proportion of CD8+ T-cells were able to release IFNγ and TNFα, to proliferate (Ki67 expression) and to degranulate (CD107a and Granzyme B expression) after short incubation with MHC class-I peptide. These vaccines strategies were also able to induce a higher proportion of memory CD8+ T-cells and the expression of PD-1 on CD8+ T-cells. Additionally, splenocytes from vaccinated mice were able to kill MHC class I peptide-pulsed targeT-cells (T2 and RMAS cells) and murine PCa cells expressing PAP (TRAMP-C1 cells) in vitro. The vaccine slowed the growth of human-PAP+-B16-derived tumors in HHDII/DR1 model and prolonged the survival (9 days delay for 30% of the mice). Splenic CD8+ T-cells and CD8+ tumor-infiltrating cells from vaccinated mice released IFNγ and TNFα, proliferated and degranulated in response to in vitro stimulation with MHC class I peptide. Finally, multimer-based flow cytometric analysis of peripheral blood mononuclear cells from patients with PCa following in vitro stimulation with PAP-derived peptides demonstrated the presence of PAP42mer-reactive CD8+ T-cells. Ongoing studies are assessing the effect of combining the vaccine with checkpoint inhibition (a PD-1 monoclonal antibody) in a therapeutic setting. In summary, we have developed a vaccine strategy which induces robust anti-PAP immunity and slows the growth of implanted PAP+ cancer cell-derived tumors in vivo. The presence of PAP-specific CD8+ T-cells in the periphery of patients with advanced PCa suggests that these patients will benefit from this new approach, especially when combined with checkpoint inhibition.
Citation Format: Pauline Le Vu, Jayakumar Vadakekolathu, Dennis Christensen, Lindy Durrant, Graham Pockley, Stephanie E.B. McArdle. Development of a novel prostatic acid phosphatase-derived vaccine for the treatment of advanced prostate cancer [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B118.