Fn3 proteins engineered to recognize tumor biomarker mesothelin internalize upon binding.
Fn3 proteins engineered to recognize tumor biomarker mesothelin internalize upon binding.
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DOI：
10.1371/journal.pone.0197029
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Moore SJ
中科院分区：
文献类型：
--
作者：
Sirois AR;Deny DA;Baierl SR;George KS;Moore SJ
间皮素是一种细胞表面蛋白，在许多癌症中过表达，包括乳腺癌、卵巢癌、肺癌、肝癌和胰腺肿瘤。已证明间皮素的异常表达通过与已确定的肿瘤生物标志物CA 125的相互作用促进肿瘤进展和转移。因此，特异性结合间皮素的分子具有潜在的治疗和诊断应用。然而，目前没有间皮素靶向分子被批准用于常规临床用途。虽然靶向间皮素的抗体正在开发中，但一些临床应用可能需要具有替代蛋白质折叠的靶向分子。例如，非抗体蛋白质更适合于分子成像，并且可以促进不同的化学缀合策略以产生药物递送复合物。在这项工作中，我们工程化的纤连蛋白III型结构域（Fn 3）的非抗体蛋白质支架的变体，以高亲和力结合间皮素，使用定向进化和酵母表面展示。先导工程化的Fn3变体以高产量从细菌培养物中可溶性地产生和纯化。在与人癌细胞系上的间皮素特异性结合后，工程化的Fn3蛋白内化并共定位于早期内体。据我们所知，这是第一次报告的非抗体蛋白质工程结合间皮素。结果证实，非抗体蛋白质可以被工程化以结合肿瘤生物标志物间皮素，并鼓励继续开发用于靶向诊断和治疗等应用的工程化变体。
Mesothelin is a cell surface protein that is overexpressed in numerous cancers, including breast, ovarian, lung, liver, and pancreatic tumors. Aberrant expression of mesothelin has been shown to promote tumor progression and metastasis through interaction with established tumor biomarker CA125. Therefore, molecules that specifically bind to mesothelin have potential therapeutic and diagnostic applications. However, no mesothelin-targeting molecules are currently approved for routine clinical use. While antibodies that target mesothelin are in development, some clinical applications may require a targeting molecule with an alternative protein fold. For example, non-antibody proteins are more suitable for molecular imaging and may facilitate diverse chemical conjugation strategies to create drug delivery complexes. In this work, we engineered variants of the fibronectin type III domain (Fn3) non-antibody protein scaffold to bind to mesothelin with high affinity, using directed evolution and yeast surface display. Lead engineered Fn3 variants were solubly produced and purified from bacterial culture at high yield. Upon specific binding to mesothelin on human cancer cell lines, the engineered Fn3 proteins internalized and co-localized to early endosomes. To our knowledge, this is the first report of non-antibody proteins engineered to bind mesothelin. The results validate that non-antibody proteins can be engineered to bind to tumor biomarker mesothelin, and encourage the continued development of engineered variants for applications such as targeted diagnostics and therapeutics.
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