VINE-seq

VINE-seq (Vessel Isolation and Nuclei Extraction for Sequencing) is a method to isolate and molecularly characterize the vascular and perivascular cells of the human brain microvessels at single-nuclei resolution. This technique is achieved by combining various known laboratory-based strategies involving the mechanical dissociation of brain tissue samples into single cells, density gradient centrifugation and filtration to isolate nuclei of microvessels, fluorescence-activated cell sorting (FACs) of cellular populations and droplet-based single-nuclei RNA sequencing (drop-snRNA-seq). Altogether, this generates a single-nuclei transcriptomic profile of the various cell types present in the vasculature of the brain. Through processing and analyzing the single-nuclei transcriptomic data, the heterogeneity within and between cell types can be distinguished to construct the molecular landscape of the human brain vasculature that was not previously done before.

Background

A central feature of brain health is the maintenance of the complex vasculature that functions to transport essential metabolites, oxygen, nutrients and wastes to and from the human brain. Disruption of the vasculature can lead to neurological disorders such as Alzheimer's disease and stroke. The vasculature is home to a heterogeneous group of cell types including but not limited to the endothelial cells, pericytes, astrocytes, and leukocytes. Although there have been many attempts to understand the cellular diversity in the vasculature over the years, methodological and technological limitations have hindered our ability to decode the complex entity of the brain. Therefore, our treatment options for neurological disorders have been inadequate due to our incomplete characterization of the vascular and perivascular cells in this complex environment. Through recent advances in single-cell RNA sequencing (scRNA-seq), the molecular atlas of the brain vasculature in mice has been decoded and the heterogeneous nature within the cell types found in the human brain have been slowly uncovered.