Abstract 3391: Companion diagnostic strategies specific to antibody therapies
Authors: Krueger Joseph S. , Young David , Lange Holger , Potts Steve 
Published: August 2015

Abstract
One premise of antibody-drug conjugates (ADC) is that the bound mAb-antigen complex on the cell surface will internalize and be metabolized by lysosomal proteases to release the free drug. Thus, the efficacy of an ADC is dependent not only on the presence of cell surface antigens, but also an active system of receptor turnover and receptor-mediated endocytosis. Thus, a predictive assay for patient response would ideally account for both the degree of cell surface expression of the target, as well as cytoplasmic presence of the target to quantify a surrogate for receptor turnover and internalization. Immunohistochemistry based assays (IHC) are best suited to address these questions, as it is the only method which provides the ability to measure both membrane and cytoplasm expression of the target simultaneously within archival FFPE biopsies. However, the biological mechanisms behind receptor internalization and turnover have not been elucidated for novel therapeutic targets. In most cases, an IHC assay is utilized to evaluate these measures, without prior advance knowledge of how these measures are suitable for patient selection. Unanticipated difficulties in tissue interpretation, such as low apparent expression of the target, occlusion of membrane staining by cytoplasmic staining, or heterogeneity in staining often lead to failure in determining a correct patient stratification approach.
In order to investigate patient selection strategies for ADCs, we have invented several proprietary approaches for measuring critical properties of the therapeutic target on the cell surface or inside the cell which can be used to understand and predict efficacy to an ADC using FFPE biopsies. These quantitative pathology approaches are based on image analysis approaches which been designed specifically for ADC CDx programs to develop a pathology based scoring system which can be predictive of ADC response: 1) Accurately quantifying low levels of cell surface target expression; 2) Defining cell surface target expression independent of cytoplasmic expression; 3) Overcoming staining heterogeneity; and 4) Determining the correct staining thresholds for quantification. These image analysis based approaches can be used to define and evaluate a scoring approach, train pathologists, assess objective performance, and best determine a cutpoint approach using statistical approaches. These image analysis based tools can be used to create a manual scoring paradigm for an IHC assay or can be incorporated into a medical device directly to support the PMA effort. Incorporation of these novel tools will enable ADC developers to create efficacy and patient stratification paradigms which incorporate the critical biological endpoints unique to ADCs.
Citation Format: Joseph S. Krueger, David Young, Holger Lange, Steve Potts. Companion diagnostic strategies specific to antibody therapies. [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 3391. doi:10.1158/1538-7445.AM2015-3391