Targeting a proteolytic neoepitope on CUB domain containing protein 1 (CDCP1) for RAS-driven cancers.
Targeting a proteolytic neoepitope on CUB domain containing protein 1 (CDCP1) for RAS-driven cancers.
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DOI：
10.1172/jci154604
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发表时间：
2022-02-15
期刊：
影响因子：
--
通讯作者：
Wells JA
中科院分区：
文献类型：
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作者：
Lim SA;Zhou J;Martinko AJ;Wang YH;Filippova EV;Steri V;Wang D;Remesh SG;Liu J;Hann B;Kossiakoff AA;Evans MJ;Leung KK;Wells JA
细胞外蛋白水解在疾病中经常失调，并且可以产生具有健康组织中未发现的独特新表位的蛋白质形式。在这里，我们证明选择性识别包含蛋白 1 (CDCP1) 的 CUB 结构域上的蛋白水解新表位的抗体可以为实体瘤提供更有效、更安全的治疗。 CDCP1 在 RAS 驱动的癌症中高度过表达，其胞外域被细胞外蛋白酶切割。生化、生物物理和结构表征表明，CDCP1 的 2 个切割片段仍然与最小的蛋白水解诱导的构象变化紧密相关。使用差异噬菌体展示，我们生成了重组抗体，该抗体对切割的CDCP1具有出色的选择性，并且与未切割的形式没有可检测到的结合。这些抗体作为抗体-药物缀合物、抗体-放射性核素缀合物和双特异性 T 细胞接合剂，有效靶向表达 CDCP1 的裂解癌细胞。在同基因胰腺肿瘤模型中，这些切割特异性抗体显示出肿瘤特异性定位和抗肿瘤活性，与泛 CDCP1 方法相比，具有更高的安全性。靶向蛋白水解新表位可以提供正交“AND”门来提高治疗指数。
Extracellular proteolysis is frequently dysregulated in disease and can generate proteoforms with unique neoepitopes not found in healthy tissue. Here, we demonstrate that Abs that selectively recognize a proteolytic neoepitope on CUB domain containing protein 1 (CDCP1) could enable more effective and safer treatments for solid tumors. CDCP1 is highly overexpressed in RAS-driven cancers, and its ectodomain is cleaved by extracellular proteases. Biochemical, biophysical, and structural characterization revealed that the 2 cleaved fragments of CDCP1 remain tightly associated with minimal proteolysis-induced conformational change. Using differential phage display, we generated recombinant Abs that are exquisitely selective to cleaved CDCP1 with no detectable binding to the uncleaved form. These Abs potently targeted cleaved CDCP1-expressing cancer cells as an Ab-drug conjugate, an Ab-radionuclide conjugate, and a bispecific T cell engager. In a syngeneic pancreatic tumor model, these cleaved-specific Abs showed tumor-specific localization and antitumor activity with superior safety profiles compared with a pan-CDCP1 approach. Targeting proteolytic neoepitopes could provide an orthogonal “AND” gate for improving the therapeutic index.