Abstract 6412: Development of a high-throughput assay platform for the screening of selective integrin inhibitors in cancer

Abstract Integrins are critical mediators of cancer progression, facilitating cancer cell invasion, survival in circulation, and metastatic outgrowth. Integrins result in cancer progression by activating pro-survival signaling pathways and remodeling the cancer microenvironment, and thus represent promising targets for anticancer treatments. To accelerate the discovery of novel integrin-targeted therapeutics, we developed an integrated platform of high-throughput screening assays for the evaluating integrin inhibitors of diverse modalities, including small molecules, peptides, antibodies, and antibody-drug conjugates. In this study, we developed a panel of cell-free fluorescence polarization (FP) assays in 384-well plate format using a Cy3B-RGD probe to screen compounds against multiple RGD-binding integrins (αvβ1, αvβ3, αvβ5, αvβ6, αvβ8 and α8β1). In addition, we generated HEK293 cell lines stably overexpressing integrins such as α8β1, and developed ELISA as well as IncuCyte based α8β1-Mfge8 competition binding assays for compounds screening in cell-based assays. The assay results were further validated using cancer cell lines endogenously expressing high levels of integrins such as α8β1, confirming compound binding and activity in a more physiologically relevant model. In summary, we have created a robust high-throughput screening platform that combines cell-free and cell-based assays. This integrated integrin assay platform enables the efficient evaluation of selective integrin inhibitors, accelerating the development of novel integrin inhibitors for target-based cancer treatment. Citation Format: Ziwei Zhang, Yu Zhou, Peichuan Zhang, Zhonghua Yan. Development of a high-throughput assay platform for the screening of selective integrin inhibitors in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 6412.