Dynamical rearrangement of human epidermal growth factor receptor 2 upon antibody binding: effects on the dimerization
Authors: Magalhães Pedro R. , Machuqueiro Miguel , Almeida José G. , Melo André , Cordeiro M. Natalia D. S. , Verde Sandra Cabo , Gümüş Zeynep H. , Moreira Irina S. , Correia João D. G. , Melo Rita 
Published: September 2019

AbstractHuman epidermal growth factor 2 (HER2) is a ligand-free tyrosine kinase receptor of the HER family that is overexpressed in some of the most aggressive tumours. Treatment of HER2+ breast cancers with the humanized monoclonal anti-HER2 antibody (Trastuzumab) revealed highly effective, encouraging the development of various HER2-specific antibodies, kinase inhibitors and dimerization inhibitors for cancer therapy. Although it is known that HER2 dimerization involves a specific region of its extracellular domain, the so-called “dimerization arm”, the mechanism of dimerization inhibition remains uncertain. However, uncovering how antibody interactions lead to inhibition of HER2 dimerization is of key importance in understanding its role in tumour progression and therapy. Herein, we employed several computational modelling techniques for a molecular-level understanding of the interactions between HER and specific anti-HER2 antibodies, namely an antigen-binding (Fab) fragment (F0178) and a single chain variable fragment from Trastuzumab (scFv). Specifically, we investigated the effects of antibody-HER2 interactions on the key residues of “dimerization arm” from molecular dynamics (MD) simulations of unbound HER (in a total of 1 µs), as well as scFv:HER2 and F0178:HER2 complexes (for a total of 2.5 µs). A deep surface analysis of HER receptor revealed that the binding of specific anti-HER2 antibodies induced conformational changes both in the interfacial residues, which was expected, and in the ECDII, in particular at the “dimerization arm”, which is critical in establishing protein-protein interface (PPI) interactions. Our results support and advance the knowledge on the already described trastuzumab effect on blocking HER2 dimerization through synergistic inhibition and/or steric hindrance. Furthermore, our approach offers a new strategy for fine-tuning target activity through allosteric ligands.Author summaryIncreasing insight into the genetics and molecular biology of diseases has resulted in the identification of a high number of potential molecular targets for drug discovery and development. Human Epidermal Growth Factor Receptor 2 (HER2) is one of the most relevant Epidermal Growth Factor Receptor (EGFR) members, whose overexpression has been shown to play an important role in the development and progression of certain aggressive types of breast cancer. Thus, the development of novel approaches on anti-HER2 therapies is quiet relevant. Molecular modelling and simulation can be used to bring new perspectives, both in structure-based drug design and to provide atomistic information of the intermolecular coupling dynamics between inhibitors and receptors via interactive computographic software. Considering my interest in drug design and mechanisms of action using integrated in silico approaches, herein, I have used multiple methods to evaluate how HER2 coupling to its different partners could alter this functional mechanism. The results suggest that the antibodies fragments studied show different dynamic complexes with HER2 although both could contribute to downstream of the tumour cell receptor pathways. I do believe that future research breakthroughs with aid of chemo-bioinformatics will allow a more comprehensive perception on biomedicine.