Abstract 1733: MP0317, a CD40xFAP targeting multi-specific DARPin® therapeutic, drives immune activation and reverts myeloid-mediated T-cell suppression in vitro and ex vivo
Authors: Ioannou Kyriaki , Ragusa Simone , Roquette Joanna , Florescu Ana , Gachechiladze Mariam , Müller Eliane , Martinez-Gomez Julia M. , Barsin Sophie , Jetzer Sarah , Rigamonti Nicolo , Domke Clara , Lekishvili Tamara , Rommel Karolin , Tosevski Ivana , Goubier Anne , Legenne Philippe , Kirkin Vladimir , Levesque Mitch , Ji Hong , Kenefeck Rupert 
Published: July 2021

Abstract
Therapeutic agonists targeting CD40 have shown encouraging signs of anti-tumor efficacy in patients. Despite their initial promise however, optimal dosing of systemically active agents has been limited by toxicity and their full therapeutic potential not achieved. We have previously demonstrated in vitro the ability of MP0317, a novel multi-specific DARPin® therapeutic, to selectively activate CD40 bearing B-cells, dendritic cells and macrophages in the presence of fibroblast activation protein (FAP). FAP is highly expressed in the stroma of many solid tumors and is found only at lower levels in other tissues. MP0317 utilizes FAP and CD40 binding domains to selectively cross-link and activate CD40 in the presence of FAP and thus avoid systemic CD40 activation classically associated with toxicity. Here we present new data which confirm the unique therapeutic potential of MP0317 in ex vivo model systems, demonstrate modulation of macrophage phenotype and reveal a release of T-cells from macrophage-mediated suppression. Ex vivo functional assays performed with MP0317 on dissociated human tumors demonstrated FAP-dependent activation of CD40-expressing B-cell and myeloid cell populations. Immunohistochemical analysis of tissue micro-arrays demonstrated FAP levels consistent with those required for immune cell activation in a broad range of solid tumor indications. Furthermore, by mining publicly available data, we classified tumor indications based on their immune cell content, pathway activation and MP0317 ex vivo mode of action (T cell/macrophage ratio, FAP and CD40 expression) and proposed patient populations and indications for the upcoming clinical trials. To further investigate the functional impact of MP0317, macrophages were differentiated in vitro using canonical polarising cytokines. Suppressive tumor-associated macrophage (TAM)-like cells, were repolarized by MP0317 to an anti-tumor-like phenotype. Furthermore, we explored whether modulation of macrophage phenotype by MP0317 could relieve their suppressive effect on T-cells. TAM-like macrophages potently supressed anti-CD3/28 driven T-cell activation and indeed the addition of MP0317 was found to restore T-cell response. Together these data further support MP0317's FAP targeted multimodal mechanism of action in human tumors, provide evidence for an improved therapeutic window over existing CD40 agonists and support progression to the clinic with an informed target patient profile.
Citation Format: Kyriaki Ioannou, Simone Ragusa, Joanna Roquette, Ana Florescu, Mariam Gachechiladze, Eliane Müller, Julia M. Martinez-Gomez, Sophie Barsin, Sarah Jetzer, Nicolo Rigamonti, Clara Domke, Tamara Lekishvili, Karolin Rommel, Ivana Tosevski, Anne Goubier, Philippe Legenne, Vladimir Kirkin, Mitch Levesque, Hong Ji, Rupert Kenefeck. MP0317, a CD40xFAP targeting multi-specific DARPin® therapeutic, drives immune activation and reverts myeloid-mediated T-cell suppression in vitro and ex vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1733.