Abstract P229: Neurovascular and Cognitive Dysfunction in Hypertension at the Single-Cell Level

Abstract

Hypertension (HTN) is the leading cause of vascular cognitive impairment, which has been attributed to altered microvascular regulation, neural network dysfunction and white matter (WM) damage. However, the neurovascular cells and cellular networks through which microvascular alterations lead to neuronal dysfunction and WM damage require elucidation. We employed unbiased single-cell RNA sequencing of the neocortex in 10-week-old C57BL/6 male mice treated with AngII (600ng/kg/min s.c.) or vehicle, to map out the transcriptomic changes induced by HTN. Transcriptomes (3 mice/group) were obtained for 39,451 cells before (day 3 after AngII) and after development of neurovascular and cognitive dysfunction (day 42). Profound transcriptional alterations were already seen at day 3 in venular endothelial cells (EC), GABAergic interneurons expressing neuronal nitric oxide synthase (nNOS), and oligodendrocyte precursors (OPC) (EdgeR; FDR< 0.05, logFC> 2). Gene ontology (GO) analysis (p <0.05) revealed terms consistent with senescence of venular EC, validated by immunolabeling with senescence markers (β-galactosidase, p27, p21, p16). GO also flagged terms reflecting suppression of neuronal NO synthesis and synaptic dysfunction in interneurons, which were confirmed by demonstrating reduced NMDA-evoked NO release in brain slices (20±0.03% Veh vs. 0±0.03% AngII;p<0.0001;n=60 neurons from 5 slices/group) and loss of the vesicular GABA transporter, VGAT. In addition, GO revealed early impairments in OPC differentiation, which could underlie later WM disruption. Consistent with this prediction, transcripts suggestive of impaired myelination were overrepresented at 42 days in mature oligodendrocytes (e.g., Hes5 ), and these WM alterations were confirmed in vivo by an increase in internodal distance (Nav1.6/CASPR) in axons (0.09±0.014 µm 3 Veh vs. 0.16±0.01 µm 3 AngII;n=4/group;p<0.05). Ligand-receptor interaction analysis (CellChat) indicated molecular reprogramming of oligodendrocytes by senescent EC via suppression of signaling promoting myelination (CD39-AdeRA1) and upregulation of signaling arresting OPC development (Endothelin-ER2). These data reveal a previously unappreciated susceptibility of venular EC, OPC and nNOS interneurons to the deleterious effects of AngII HTN, and point to a novel pathogenic interaction between senescent EC and OPC, as well as to early nNOS interneuron dysfunction, the disease relevance and therapeutic potential of which requires validation.