Cellular and Molecular Biotechnology, Faculty of Technology, Bielefeld University, 33615 Bielefeld, Germany.
Organic and Bioorganic Chemistry, Faculty of Chemistry, Bielefeld University, 33615 Bielefeld, Germany.
Int J Mol Sci. 2020 Dec 15;21(24):9535. doi: 10.3390/ijms21249535.
The epidermal growth factor receptor (EGFR) plays a central role in the progression of many solid tumors. We used this validated target to analyze the de novo design of EGFR-binding peptides and their application for the delivery of complex payloads via rational design of a viral vector. Peptides were computationally designed to interact with the EGFR dimerization interface. Two new peptides and a reference (EDA peptide) were chemically synthesized, and their binding ability characterized. Presentation of these peptides in each of the 60 capsid proteins of recombinant adeno-associated viruses (rAAV) via a genetic based loop insertion enabled targeting of EGFR overexpressing tumor cell lines. Furthermore, tissue distribution and tumor xenograft specificity were analyzed with systemic injection in chicken egg chorioallantoic membrane (CAM) assays. Complex correlations between the targeting of the synthetic peptides and the viral vectors to cells and in ovo were observed. Overall, these data demonstrate the potential of computational design in combination with rational capsid modification for viral vector targeting opening new avenues for viral vector delivery and specifically suicide gene therapy.
表皮生长因子受体 (EGFR) 在许多实体肿瘤的进展中起着核心作用。我们利用这个经过验证的靶点，分析了 EGFR 结合肽的从头设计及其在通过合理设计病毒载体来递送电荷的应用。通过计算设计了与 EGFR 二聚化界面相互作用的肽。化学合成了两个新的肽和一个参考肽（EDA 肽），并对其结合能力进行了表征。通过在重组腺相关病毒 (rAAV) 的每个 60 个衣壳蛋白中的遗传环插入来展示这些肽，使能够针对 EGFR 过表达的肿瘤细胞系进行靶向。此外，通过鸡胚尿囊膜 (CAM) 试验中的系统注射分析了组织分布和肿瘤异种移植特异性。观察到合成肽和病毒载体与细胞和体内的靶向之间存在复杂的相关性。总的来说，这些数据表明计算设计与合理的衣壳修饰相结合，为病毒载体靶向提供了潜力，为病毒载体递送电荷，特别是自杀基因治疗开辟了新的途径。
