Introduction: GD2 is expressed on neuroblastoma, melanomas, small cell lung cancers and sarcomas. While it is expressed minimally on normal tissues, it is expressed on nerve cells and, anti-GD2 (dinutuximab) treatment can cause neuropathic pain. To increase tumor specificity, we developed a bispecific SNIPER antibody to simultaneously target two tumor antigens (GD2 and B7H3). B7H3 is overexpressed on multiple tumor types, with minimal expression on most normal cells and is absent on nerve cells. The goal of this SNIPER is to enhance tumor-specificity, while reducing toxicity, ultimately improving efficacy for cancers expressing both targets.
Experimental Procedures: SNIPER specificity was tested by flow cytometry for binding to GD2 +/- B7H3 expressing cells. Using an Incucyte S3, we tested antibody internalization properties [compared to anti-B7H3 and dinutuximab monoclonal antibodies (mAbs)] and antibody dependent cellular-cytotoxicity (ADCC) capabilities. Mice bearing variants of GD2/B7H3-expressing tumors were intravenously injected with 89Zr- radiolabeled SNIPER and its longitudinal in vivo biodistribution was monitored via positron emission tomography imaging. In vivo efficacy studies of SNIPER were tested against mice bearing either melanoma or neuroblastoma tumors that express both GD2 and B7H3.
Summary of Data: In vitro and in vivo tumor specificity testing confirmed that SNIPER specifically targets B7H3+/GD2+ cells, but it does not bind to GD2+/B7H3- cells (which simulate nerve cells). We observed high internalization of anti-GD2 mAbs; we did not observe antibody internalization of either the SNIPER or anti-B7H3 mAbs. SNIPER was as effective at ADCC as the dinutuximab, but we saw minimal ADCC with the anti-B7H3 mAbs. An afucosylated version of SNIPER showed significantly enhanced ADCC compared to dinutuximab. Our in vivo efficacy studies found that SNIPER was as as dinutuximab when given at the same dose.
Conclusions: The Fab arms of SNIPER targeting GD2 and B7H3 each have low-to-moderate affinity. SNIPER binds with strong avidity when both arms bind to their antigens on the same cell. Stronger avidity through both arms binding results in high-tumor specificity. Because SNIPER should not bind to nerves, it may be possible to administer increased doses of SNIPER beyond the tolerable dose of dinutuximab, which could further improve efficacy. Ongoing studies include antitumor efficacy testing, nerve binding assays and assessing reduction of pain in vivo with SNIPER.
Citation Format: Amy K. Erbe, Arika Feils, Zack Rosenkrans, Jessica Wiwczar, Daniel Gerhardt, Bonnie Hammer, Mildred Felder, Mark Bercher, Alina Hampton, Lizzie Frankel, Dan Spiegelman, Noah Tsarovsky, Alexander Rakhmilevich, Jacquelyn Hank, Bryan Glaser, Reinier Hernandez, Roland Green, Paul M. Sondel. Tumor-targeting and efficacy of B7H3/GD2 bispecific SNIPER antibodies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2019.
