Peptide nanofibrils (PNFs) are gaining consideration as promising transduction enhancers to enhance viral vector supply in ex vivo gene remedy purposes. Nonetheless, the affect of PNF structural morphology on viral seize and mobile interplay stays poorly understood. Right here, we systematically evaluate two clinically related PNFs, D4, a brief β-sheet-forming peptide, and Vectofusin-1, an α-helical peptide, specializing in their fibrillar structure, viral particle binding, and interplay with host cells. Secondary construction evaluation, molecular dynamics simulations and electron microscopy revealed that D4 kinds loosely packed, β-sheet-rich aggregates, whereas Vectofusin-1 assembles into compact, α-helical constructions. Each superstructures have a optimistic and hydrophobic floor that are key determinants for interplay with viral and plasma membranes. Upon publicity to virus-like particles (VLPs), D4 aggregates grew in dimension and density, whereas Vectofusin-1 fashioned extra quite a few, smaller aggregates. D4 certain VLPs with markedly increased density, yielding a uniform virion coating, in distinction to the decrease and heterogeneous VLP affiliation noticed for Vectofusin-1. Notably, solely D4 aggregates have been actively engulfed by filopodia resulting in lively uptake through endocytosis primarily by macropinocytosis and subsequent degradation by lysosomes. In distinction, Vectofusin-1 binding to plasma membrane appeared extra passive with minimal internalization. These distinct behaviors have been maintained underneath transduction-like situations, with D4 facilitating direct VLP contact with the plasma membrane and Vectofusin-1 forming extracellular networks. Our outcomes reveal that PNF combination morphology critically determines viral and mobile interactions and counsel that D4 might provide superior efficacy and security profiles to be used in ex vivo gene therapies.
