Abstract 1324: A translational platform to design antibodies targeting triple negative breast cancer-specific antigens for cancer immunotherapy
Authors: Ilieva Kristina M. , Marlow Rebecca , Cheung Anthony , Francesch Erika , Karagiannis Panagiotis , Fittall Matthew , Crescioli Silvia , Tutt Andrew , Karagiannis Sophia 
Published: August 2015

Triple negative breast cancers (TNBCs) are characterised by the lack of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 (HER2) rendering them insensitive to current endocrine and anti-HER2 therapies, therefore novel treatment options are required. Breast cancers have been shown to be immunogenic and patient tumors feature infiltrating immune cells. Since monoclonal antibodies are an important therapeutic modality in the clinical management of many cancers, engineering antibodies against triple negative breast cancer specific antigens that may activate immune effector cells within tumors and in the circulation may lead to tumour clearance and may provide new treatment strategies for this patient group. Here we describe a platform for selection of TNBC-associated antigens, engineering of novel antibodies against these antigens and functional studies to elucidate antibody mechanisms of action and therapeutic potential in vitro and vivo. Our approach involves identification of tumour-associated antigens from patient tumor specimens using a transcriptomics approach and validation and evaluation of biological functions in breast cancer cells using cell culture models. Antibody heavy (VH) and light (VL) sequences are cloned in a dual expression vector system for recombinant expression in HEK293 cells to rapidly produce antibodies with reactivity to specific tumor antigens. The system is optimised for the seamless expression of V genes of any species including human sequences, along with human constant (C) genes of different antibody classes and subclasses. The latter permits the study of different class/subclass antibodies in order to identify those most effective in triggering immune cell activation and tumor clearance. Engineered monoclonal antibodies are subsequently tested for their antigen binding properties and capacity to activate immune effector cells against cancer cells in vitro. The therapeutic potential of selected candidates is evaluated in vivo using xenograft TNBC mouse models engrafted with components of the human immune system using adoptive transfer of healthy volunteer and patient derived peripheral blood cells. Xenograft models featuring human immune components serve to evaluate the therapeutic potential of monoclonal antibodies in the context of their capacity to redirect immune effector functions against cancer cells. Our findings demonstrate a translational pathway designed to facilitate the discovery and evaluation of novel antibody therapeutic options for triple negative breast cancer immunotherapy.

Citation Format: Kristina M. Ilieva, Rebecca Marlow, Anthony Cheung, Erika Francesch, Panagiotis Karagiannis, Matthew Fittall, Silvia Crescioli, Andrew Tutt, Sophia Karagiannis. A translational platform to design antibodies targeting triple negative breast cancer-specific antigens for cancer immunotherapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1324. doi:10.1158/1538-7445.AM2015-1324