Abstract 4263: Anti-Siglec 7 antibody displays potent anti-tumor immunity and demonstrates improved tumor control in combination with anti-PD1 in ovarian cancer
Authors: Bordoloi Devivasha , Kulkarni Abhijeet J. , Adeniji Opeyemi S. , Pampena M Betina , Bhojnagarwala Pratik S. , Zhao Shushu , Parzych Elizabeth M. , Zhang Rugang , Betts Michael R. , Abdel-Mohsen Mohamed , Weiner David B. 
Published: April 2023

Abstract
High grade serous ovarian cancer (OC) and ovarian carcinosarcoma, referred as “Cold” tumors have restricted treatment choices and being associated with high mortality. Current immune checkpoint inhibitors (CPIs) have had modest impact for OC treatment; with anti PD-1/PD-L1 single therapies reported response rates of 4-15%. Accordingly, additional approaches are important for these patients. Natural killer (NK) cells represent an important subset of effector lymphocytes with strategic approaches for maximizing NK reactivity to target OC are likely important. Of interest, an inhibitory receptor and glyco immune checkpoint- Siglec 7 is expressed on 100% of peripheral blood and umbilical cord NK cells. We have generated antibodies against Siglec 7 through an optimized human Siglec 7 DNA/protein immunization approach in humanized next generation transgenic mice. By high throughput flow screening, we identified several Siglec 7 antibodies and down selected relevant clones. A unique clone DB7.2 was highly specific for Siglec 7. We demonstrated the ability of DB7.2 (human IgG1) to activate NK cells and induce OC cell killing using xCELLigence RTCA for in vivo OC challenge studies. DB7.2 (1μg/ml) bound to &gt;90% of NK cells including both CD56dim and CD56bright subsets, evaluated in the PBMCs of multiple donors. It showed strong binding to recombinant Siglec 7 and Siglec 7 transduced HEK293T but not to wild type 293T cells supporting specificity. DB7.2 induced specific killing of multiple OC lines (OVCAR3&10,TOV21G, CaOV3, OVISE, PEO4) carrying different mutations; BRCA1&2, AKT, TP53, PIK3CA, BRAF etc. and resistant against a wide array of cancer drug targets; HSP90, HDAC, MTORC, DNA alkylating agents, EGFR, PARP, PI3K, and WEE1. Tumor killing was indicated to be mediated via enhanced secretion of soluble Fas, perforin, granulysin as well as granzyme A. Of note, OVISE (BRCA1mutated) and PEO4 (BRCA2 mutated, PARPi resistant) were susceptible to DB7.2 killing with EC50- 82.67 and 68.67 nM respectively. A single dose of in vivo expressed DB7.2 significantly reduced the tumor burden in an OVISE challenged humanized mice model enhancing median survival by 57 days. As OCs are highly diverse in nature and likely to require combinatorial approaches for simultaneous targeting of immune pathways; we combined DB7.2 with anti-PD1 for further investigation. In xCELLigence assay, anti-PD1 demonstrated killing of PEO4 cells with EC50 680 nM. The combination of anti PD-1 with DB7.2 showed further enhancement of OC killing in the presence of human PBMCs. This is the first demonstration of the impact of Siglec 7 targeting mAb alone as well as in combination with anti-PD1 (NK and T cell CPIs) studied for targeting OC or any human tumor. These studies have important implications for tumor therapy and provide a novel non T cell CPI potential approach to augment current immune therapy strategies.
Citation Format: Devivasha Bordoloi, Abhijeet J. Kulkarni, Opeyemi S. Adeniji, M Betina Pampena, Pratik S. Bhojnagarwala, Shushu Zhao, Elizabeth M. Parzych, Rugang Zhang, Michael R. Betts, Mohamed Abdel-Mohsen, David B. Weiner. Anti-Siglec 7 antibody displays potent anti-tumor immunity and demonstrates improved tumor control in combination with anti-PD1 in ovarian cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4263.