Kurmann, Lisa and Okoniewski, Michal and Ogunshola, Omolara O. and Leeners, Brigitte and Imthurn, Bruno and Dubey, Raghvendra K.
(2021)
Transcryptomic Analysis of Human Brain-Microvascular Endothelial Response to -Pericytes: Cell Orientation Defines Barrier Function.
Cells, 10 (4).
p. 963.
ISSN 2073-4409
Abstract
Pericytes facilitate blood–brain barrier (BBB) integrity; however, the mechanisms involved remain unclear. Hence, using co-cultures of human cerebral microvascular endothelial cells (ECs) and vascular pericytes (PCs) in different spatial arrangements, as well as PC conditioned media, we investigated the impact of PC-EC orientation and PC-derived soluble factors on EC barrier function. We provide the first evidence that barrier-inducing properties of PCs require basolateral contact with ECs. Gene expression analysis (GEA) in ECs co-cultured with PCs versus ECs alone showed significant upregulation of 38 genes and downregulation of 122 genes. Pathway enrichment analysis of modulated genes showed significant regulation of several pathways, including transforming growth factor-β and interleukin-1 regulated extracellular matrix, interferon and interleukin signaling, immune system signaling, receptor of advanced glycation end products (RAGE), and cytokine–cytokine receptor interaction. Transcriptomic analysis showed a reduction in molecules such as pro-inflammatory cytokines and chemokines, which are known to be induced during BBB disruption. Moreover, cytokine proteome array confirmed the downregulation of key pro-inflammatory cytokines and chemokines on the protein level. Other molecules which influence BBB and were favorably modulated upon EC-PC co-culture include IL-18 binding protein, kallikrein-3, CSF2 CSF3, CXCL10, CXCL11 (downregulated) and IL-1-R4; HGF, PDGF-AB/BB, PECAM, SERPIN E1 (upregulated). In conclusion, we provide the first evidence that (1) basolateral contact between ECs and PCs is essential for EC barrier function and integrity; (2) in ECs co-cultured with PCs, the profile of BBB disrupting pro-inflammatory molecules and cytokines/chemokines is downregulated; (3) PCs significantly modulate EC mechanisms known to improve barrier function, including TGF-β regulated ECM pathway, anti-inflammatory cytokines, growth factors and matrix proteins. This human PC-EC co-culture may serve as a viable in vitro model for investigating BBB function and drug transport.
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Details
| Item Type: |
Article
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| Status: |
Published |
| Creators/Authors: |
| Creators | Email | Pitt Username | ORCID  |
|---|
| Kurmann, Lisa | | | | | Okoniewski, Michal | | | | | Ogunshola, Omolara O. | | | | | Leeners, Brigitte | | | | | Imthurn, Bruno | | | | | Dubey, Raghvendra K. | | | |
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| Date: |
20 April 2021 |
| Date Type: |
Publication |
| Journal or Publication Title: |
Cells |
| Volume: |
10 |
| Number: |
4 |
| Publisher: |
MDPI AG |
| Page Range: |
p. 963 |
| DOI or Unique Handle: |
10.3390/cells10040963 |
| Schools and Programs: |
School of Medicine > Pharmacology and Chemical Biology |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
pericyte, hCMEC/D3, co-culture, blood–brain barrier, orientation, transcriptome |
| ISSN: |
2073-4409 |
| Official URL: |
http://dx.doi.org/10.3390/cells10040963 |
| Funders: |
Swiss National Sciences Foundation, Uniscientia |
| Article Type: |
Research Article |
| Date Deposited: |
09 Jul 2021 18:55 |
| Last Modified: |
09 Jul 2021 18:55 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/41413 |
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