Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States.
Chemistry and Chemical Biology Graduate Program, University of California, San Francisco, San Francisco, United States.
Elife. 2018 Jan 23;7:e31098. doi: 10.7554/eLife.31098.
While there have been tremendous efforts to target oncogenic RAS signaling from inside the cell, little effort has focused on the cell-surface. Here, we used quantitative surface proteomics to reveal a signature of proteins that are upregulated on cells transformed with KRAS, and driven by MAPK pathway signaling. We next generated a toolkit of recombinant antibodies to seven of these RAS-induced proteins. We found that five of these proteins are broadly distributed on cancer cell lines harboring RAS mutations. In parallel, a cell-surface CRISPRi screen identified integrin and Wnt signaling proteins as critical to RAS-transformed cells. We show that antibodies targeting CDCP1, a protein common to our proteomics and CRISPRi datasets, can be leveraged to deliver cytotoxic and immunotherapeutic payloads to RAS-transformed cancer cells and report for RAS signaling status in vivo. Taken together, this work presents a technological platform for attacking RAS from outside the cell.
虽然人们已经做出了巨大的努力来从细胞内部靶向致癌性 RAS 信号，但很少有努力集中在细胞表面。在这里，我们使用定量表面蛋白质组学来揭示一组在被 KRAS 转化的细胞中上调的蛋白质的特征，这些蛋白质由 MAPK 途径信号驱动。接下来，我们生成了七种这些 RAS 诱导蛋白的重组抗体工具包。我们发现，这五种蛋白质在携带 RAS 突变的癌细胞系中广泛分布。与此同时，细胞表面 CRISPRi 筛选鉴定出整合素和 Wnt 信号蛋白对于 RAS 转化细胞至关重要。我们表明，针对 CDCP1 的抗体（我们的蛋白质组学和 CRISPRi 数据集共有的一种蛋白质）可用于将细胞毒性和免疫治疗有效载荷递送至 RAS 转化的癌细胞，并报告体内 RAS 信号状态。总之，这项工作提出了一种从细胞外部攻击 RAS 的技术平台。
