Hyaluronic acid alters vessel behavior in CXCL12-treated HUVECs

Abstract

Hyaluronic acid (HA) is a key component of the extracellular matrix known for absorbing water, swelling, and altering solid stress of tumors. HA’s anionic behavior may provide important biochemical effects toward tumor progression as well. Tumors obtain nutrients by relying on signaling molecules such as CXCL12 to recruit blood vessels and promote vessel leakage. Recent work suggests that additional positively-charged residues on CXCL12’s β and γ isoforms cause different biochemical functionality compared to the well-studied α isoform. These studies aimed to determine whether the presence of HA in a tumor’s microenvironment could alter the relative response strength of CXCL12’s various isoforms on blood vessel sprouting and apparent vascular permeability. The vessel microenvironment was modeled using a 3-channel microfluidic device with Human Umbilical Vein Endothelial Cells (HUVECs) in the outer channels forming monolayers against a 3D collagen or collagen/HA matrix in the center channel. HUVECs were cultured with media containing recombinant CXCL12 (α, β or γ). Results show that total HUVEC sprouting area follows an α>β>γ trend in isoform-treated HUVECs within a collagen matrix, matching the binding affinity order of CXCL12 to endothelial CXCR4 receptors. The presence of HA decreased overall sprouting response but shifted pro-angiogenic potency towards CXCL12’s γ isoform. Vascular permeability studies also showed an α>β>γ trend for HUVECs in collagen. With HA added, control and α-treated HUVECs became less permeable while γ-treated HUVECs became more permeable. Overall results suggest that an HA-infused collagen matrix facilitates γ isoform binding, leading to a stronger isoform-specific vessel response. Knowing how HA impacts CXCL12 isoform potency on vessels will help in the future design of CXCL12-targeted cancer therapies.