A human cell type similar to murine central nervous system perivascular fibroblasts

Jianping Liu,Liqun He,Lars Muhl,Giuseppe Mocci,Sonja Gustavsson,Byambajav Buyandelger,Michael Vanlandewijck,Christer Betsholtz,Bengt Westermark,Johanna Andrae

doi:10.1016/j.yexcr.2021.112576

Abstract

The brain vasculature has several specific features, one of them being the blood-brain barrier (BBB), which supports and protects the brain by allowing for the passage of oxygen and nutrients, while at the same time preventing passage of pathogens and toxins. The BBB also prevents efficient delivery of drugs to the brain, e.g. for treatment of brain tumors. In the murine brain, perivascular fibroblasts were recently identified as a novel potential constituent of the BBB. Here we present the existence of human cells that could be the equivalent to the murine brain perivascular fibroblasts. Using RNA sequencing, we show a similar transcriptomic profile of cultured human brain cells and murine perivascular fibroblasts. These data open up a window for new hypotheses on cell types involved in human CNS diseases.

Highlights

Blood vessels in the central nervous system (CNS) are organized to support and protect our brains
We and others have identified fibroblast (FB)-like cells in the brain using singlecell RNA sequencing. Based on their expression of plateletderived growth factor receptor-alpha (Pdgfra), FB-like cells were local ized between the vascular smooth muscle cells (VSMC) and astrocytic end-foot layers, i.e. within the paravascular Virchow-Robin spaces sur rounding arteries/arterioles and veins/venules [1]
Pdgfra+ and Col1a1+ perivascular stromal cells in mouse brains have been suggested to contribute to fibrotic scar formation after spinal cord injury [2], experimental stroke [3], multiple sclerosis [4] and experimental autoimmune encephalitis [5]

Summary

Introduction

Blood vessels in the central nervous system (CNS) are organized to support and protect our brains. One of the cell lines (U787 CG) was used to establish a bioassay for the purification of PDGF [9] These cells were of unknown origin, but the hypothesis was that they were derived from glial cells [6]. Human brain vascular pericyte (#1200, Science Cell Research Laboratory, CA, USA) We speculated that these cells are the human equivalent to the per ivascular FB that we and others had found in the mouse brain. To address this question, we performed bulk-RNA sequencing of seventeen of these human cell lines following their re-establishment in culture after 50 years’ storage in liquid nitrogen, and compared their transcriptional profiles with scRNAseq data of adult murine brain vascular cells (http://betsholtzlab.org/VascularSingleCells/database.html) [1]. Our results support the hypothesis of a human brain cell type with a similar gene expression signature as murine perivascular FB-like cells

Cell lines

Bulk RNA sequencing

Bulk RNA seq data analysis

Single cell RNA seq data analysis

Establishment of cell lines