The plasma membrane is enriched for receptors and signaling proteins that are accessible from the extracellular space for pharmacological intervention. Here we conducted a series of CRISPR screens using human cell surface proteome and integrin family libraries in multiple cancer models. Our results identified ITGAV (integrin αV) and its heterodimer partner ITGB5 (integrin β5) as the essential integrin α/β pair for cancer cell expansion. High-density CRISPR gene tiling further pinpointed the integral pocket within the β-propeller domain of ITGAV for integrin αVβ5 dimerization. Combined with in silico compound docking, we developed a CRISPR-Tiling-Instructed Computer-Aided (CRISPR-TICA) pipeline for drug discovery and identified Cpd_AV2 as a lead inhibitor targeting the β-propeller central pocket of ITGAV. Cpd_AV2 treatment led to rapid uncoupling of integrin αVβ5 and cellular apoptosis, providing a unique class of therapeutic action that eliminates the integrin signaling via heterodimer dissociation. We also foresee the CRISPR-TICA approach to be an accessible method for future drug discovery studies.

质膜富含受体和信号蛋白，这些受体和信号蛋白可从细胞外空间进入以进行药物干预。在这里，我们使用人类细胞表面蛋白质组和整合素家族文库在多种癌症模型中进行了一系列 CRISPR 筛选。我们的结果确定 ITGAV（整合素 αV）及其异二聚体伙伴 ITGB5（整合素 β5）是癌细胞扩张所必需的整合素 α/β 对。高密度 CRISPR 基因平铺进一步精确定位了 ITGAV β 螺旋桨结构域内整合素 αVβ5 二聚化的整合口袋。结合计算机模拟化合物对接，我们开发了用于药物发现的 CRISPR-Tiling 指导计算机辅助 (CRISPR-TICA) 流程，并确定 Cpd_AV2 是针对 ITGAV β 螺旋桨中央口袋的先导抑制剂。 Cpd_AV2 治疗导致整合素 αVβ5 快速解偶联和细胞凋亡，提供一类独特的治疗作用，通过异二聚体解离消除整合素信号传导。我们还预计 CRISPR-TICA 方法将成为未来药物发现研究的一种可行方法。