Vascular Basement Membrane Laminins Modulate Functional Zonation of Cerebral Microvessels

Abstract

We investigated whether vascular basement membrane (BM) laminins influence vascular zonation by performing single-cell RNA sequencing on cerebral blood vessels from mice lacking the major vascular laminins in endothelial and smooth muscle BMs, laminin 4 (Lama4-/-) and laminin 5 (Tek-cre:Lama5-/-), and wild-type littermates. Our dataset expands existing cerebral vascular transcriptomic profiles and reveals that Lama4-/- endothelial cells exhibit increased arterial marker expression and reduced postcapillary venule identity. In vitro and in vivo studies indicate that compensatory upregulation of laminin 5 in Lama4-/-vessels enhances expression of junctional proteins (Ocln, Cldn5) and promotes vessel contractility via increased expression of contractile molecules in mural cells. Additionally, loss of Lama4 upregulates expression of large artery markers (Gja4, Dll4, Tgfb2) and results in elevated autotaxin (Enpp2) levels, a key enzyme in lysophosphatidic acid production implicated in stroke. Accordingly, Lama4-/- mice exhibit worsened stroke outcomes, driven not by immune infiltration or junctional defects, but by increased vascular permeability likely mediated by autotaxin and/or activation of resident myeloid cells. Our data suggest that laminin 4/5 ratios in vascular BMs regulate functional zonation between arterioles, capillaries and postcapillary venules by modulating metabolic pathways in endothelial and mural cells, and indirectly influencing resident myeloid cells.