Heparin flexibility within the extracellular matrix determines the bioactivity of bound vascular endothelial growth factor

Vascular endothelial growth factor (VEGF), a major regulator of blood vessel formation, is naturally bound to heparan sulfate proteoglycans in the extracellular matrix (ECM). Yet, how the physical presentation of VEGF by the matrix impacts its signaling potential remains fully unknown. To address this question, we have developed a tunable heparin-containing hydrogel model that recapitulates natural VEGF binding modes with full and independent control over physical properties. Using this model, we show that the degree of heparin flexibility in the hydrogel network determines the mobility of bound VEGF and, in turn, its ability to interact with VEGF receptor 2. We demonstrate that VEGF mobility is driven by a relay mechanism, in which VEGF molecules directly switch from one heparin chain to another. When strong electrostatic interactions between heparin and other hydrogel constituents immobilize the sugar backbone, this relay mechanism is impeded, in turn reducing VEGF’s ability to reach its target receptor at the cell membrane, thereby reducing its bioactivity. This work identifies heparin flexibility within the ECM as a previously unknown regulator of the microenvironment, which will not only contribute to a better mechanistic understanding of how the ECM regulates growth factor bioactivity, but it will also provide an important design criterion for the development of tissue-engineered biomaterials that require vascularization.