Background and Purpose: Although immunotherapy as an adjuvant to surgery and chemotherapy has been investigated in ovarian cancer (OC) as a means of reducing tumor recurrence and improving survival, there remains a significant unmet need for combinatorial strategies to enhance the antitumor immune response. The purpose of this study was to develop a novel combination immunotherapy for OC, utilizing our novel fusion protein to target and generate a cellular immune response to mesothelin (MSLN) in conjunction with blockade of the PD-1/PD-L1 checkpoint to restore the function of cytotoxic T cells in order to enhance cancer control and prolong survival.

Experimental Procedures: Luciferase-expressing ID8 cells were employed to establish an intraperitoneal ovarian tumor model in immunocompetent C57BL/6 mice. The efficacies of the MSLN-targeted immune-activating fusion protein (VIC-008), αPD-L1, and the combination were evaluated. Mice received 4 intraperitoneal (i.p.) treatments of VIC-008 from day 7 post tumor inoculation weekly, and 6 treatments of αPD-L1 i.p. every other day from 4 weeks post inoculation. Tumor growth was monitored by in vivo imaging of luciferase activity. Survival time was calculated as life span from the day of tumor inoculation. In immunological studies, mice were sacrificed 7 weeks after tumor cell inoculation. Immune cells from lymph nodes, ascites and tumors were stained with antibodies against multiple immune cell markers and profiled by flow cytometry.

Results: VIC-008, αPD-L1 or combination treatment delayed tumor growth. The combination treatment resulted in the greatest prolongation in survival, followed by αPD-L1 treatment and then VIC-008 treatment. Improved survival was associated with increased levels of intratumoral CD3+CD8+ T cells (P<0.0001). The combination treatment also reduced the proportion of CD4+CD25+Foxp3+ Treg cells (P<0.0001) in the lymph nodes. An increased number of CD8+CD27+CD44+ memory T cells (P=0.0134) were observed in ascites in the combination treatment group. CD11b+CD11c+ dendritic cells were enriched in ascites in VIC-008 treatment (P=0.0019) and combination treatment groups (P=0.0010). More CD11c+CD38+ (M1) (P=0.0361) and fewer CD206+CD106+ (M2) (P=0.0285) macrophages were found in the tumors of the combination treatment group.

Conclusion: Our results suggest that, through activating dendritic cells and enhancing antigen presentation and cross-presentation, VIC-008 augments antitumor CD8+ T cell responses and facilitates generation of memory T cells when combined with PD-1/PD-L1 blockade, providing long-term antitumor effects. Our findings demonstrate for the first time a mechanistic rationale for combining VIC-008 and αPD-L1 in treatment of OC in mice, positioning this combination therapy as a potential promising new immunotherapeutic approach for OC.

Citation Format: Yang Zeng, Binghao Li, Qiuyan Liu, Patrick Reeves, Ann Sluder, Jeffrey Gelfand, Timothy Brauns, Mark Poznansky, Huabiao Chen. Combination of mesothelin-targeted immune-activating fusion protein and anti-PD-L1 augments antitumor immunity and prolongs survival in murine model of ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3649. doi:10.1158/1538-7445.AM2017-3649