EGFR-Binding Peptides: From Computational Design towards Tumor-Targeting of Adeno-Associated Virus Capsids

• Published in: International journal of molecular sciences 2020-12, Vol.21 (24), p.9535
• Main Authors: Feiner, Rebecca C, Kemker, Isabell, Krutzke, Lea, Allmendinger, Ellen, Mandell, Daniel J, Sewald, Norbert, Kochanek, Stefan, Müller, Kristian M
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Binding; Binding sites; Capsid - chemistry; Capsid - metabolism; Capsid Proteins - chemistry; Capsid Proteins - metabolism; Capsids; Cell division; Cell Line, Tumor; Chick Embryo; Chorioallantoic membrane; Chorioallantoic Membrane - metabolism; Circular Dichroism; Computational Biology; Computer applications; cyclic peptide; Dependovirus - chemistry; Dependovirus - metabolism; Dimerization; Enzymes; Epidermal growth factor; Epidermal growth factor receptors; ErbB Receptors - chemistry; ErbB Receptors - genetics; ErbB Receptors - metabolism; Gene therapy; Genetic Therapy; Genetic Vectors; Growth factors; Heparan sulfate; Humans; Hydrogen bonds; Hydrophobic surfaces; Insertion; Kinases; Ligands; Microscopy, Fluorescence; Mutation; oncolytic virus; Oncolytic Viruses - chemistry; Oncolytic Viruses - genetics; Oncolytic Viruses - metabolism; Payloads; Peptides; Peptides - chemical synthesis; Peptides - chemistry; Protein Binding; protein engineering; Protein Engineering - methods; Regulatory approval; Solid tumors; Suicide; Suicide genes; synthetic biology; Synthetic peptides; Transplantation, Heterologous; Tumor cell lines; Tumors; Up-Regulation; VDEPT; Viruses; VP protein; Wound Healing - drug effects; Xenografts; Xenotransplantation
• ISSN: 1422-0067; 1661-6596; 1422-0067
• 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.