De Novo Design of Integrin a5ß1 Modulating Proteins for Regenerative Medicine

Integrin a5ß1 is crucial for cell attachment and migration in development and tissue regeneration, and a5ß1 binding proteins could have considerable utility in regenerative medicine and next-generation therapeutics. We use computational protein design to create de novo a5ß1-specific modulating miniprotein binders, called NeoNectins, that bind to and stabilize the open state of a5ß1. When immobilized onto titanium surfaces and throughout 3D hydrogels, the NeoNectins outperform native fibronectin and RGD peptide in enhancing cell attachment and spreading, and NeoNectin-grafted titanium implants outperformed fibronectin and RGD-grafted implants in animal models in promoting tissue integration and bone growth. NeoNectins should be broadly applicable for tissue engineering and biomedicine. Overall design: MCF 10A, MSC, hOBS, and hFF cells were treated with integrin a5 B1 agonists or antagonists on plastic or titatnium surfaces to compare the effects of the de novo binders on cell transcriptomes