Transcriptome analysis reveals microvascular endothelial cell-dependent pericyte differentiation

Brandt, Maarten M.; Dijk, Christian G.M. van; Maringanti, Ranganath; Chrifi, Ihsan; Kramann, Rafael; Verhaar, Marianne C.; Duncker, Dirk J.; Mokrý, Michal; Cheng, Caroline

doi:10.1038/s41598-019-51838-x

Cited by 25 publications

(25 citation statements)

References 54 publications

(59 reference statements)

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“…To verify the characteristics of PCs used, we investigated the expression of commonly applied PC markers at the transcriptional level [ 8 , 15 , 22 ] ( Figure 4 a). Several marker genes were highly expressed in our PCs, including platelet-derived growth factor receptor-beta (PDGFR-β), alpha-smooth muscle actin (ACTA2), vimentin (VIM), regulator of G-protein signaling (RGS5), amino peptidase N (ANPEP) and non-muscle myosins (MYH9/10).…”

Section: Resultsmentioning

confidence: 99%

Transcriptomic Analysis of Human Brain -Microvascular Endothelial Cell Driven Changes in -Vascular Pericytes

Kurmann

Okoniewski

Dubey

2021

Cells

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Many pathological conditions of the brain are associated with structural abnormalities within the neurovascular system and linked to pericyte (PC) loss and/or dysfunction. Since crosstalk between endothelial cells (ECs) and PCs greatly impacts the function of the blood–brain barrier (BBB), effects of PCs on endothelial integrity and function have been investigated extensively. However, the impact of ECs on the function and activity of PCs remains largely unknown. Hence, using co-cultures of human brain vascular PCs with human cerebral microvascular ECs on opposite sides of porous Transwell inserts which facilitates direct EC–PC contact and improves EC barrier function, we analyzed EC-driven transcriptomic changes in PCs using microarrays and changes in cytokines/chemokines using proteome arrays. Gene expression analysis (GEA) in PCs co-cultured with ECs versus PCs cultured alone showed significant upregulation of 1′334 genes and downregulation of 964 genes. GEA in co-cultured PCs revealed increased expression of five prominent PC markers as well as soluble factors, such as transforming growth factor beta, fibroblast growth factor, angiopoietin 1, brain-derived neurotrophic factor, all of which are involved in EC–PC crosstalk and BBB induction. Pathway enrichment analysis of modulated genes showed a strong impact on many inflammatory and extracellular matrix (ECM) pathways including interferon and interleukin signaling, TGF-β and interleukin-1 regulation of ECM, as well as on the mRNA processing pathway. Interestingly, while co-culture induced the mRNA expression of many chemokines and cytokines, including several CCL- and CXC-motif ligands and interleukins, we observed a decreased expression of the same inflammatory mediators on the protein level. Importantly, in PCs, ECs significantly induced interferon associated proteins (IFIT1, IFI44L, IF127, IFIT3, IFI6, IFI44) with anti-viral actions; downregulated prostaglandin E receptor 2 (prevent COX-2 mediated BBB damage); upregulated fibulin-3 and connective tissue growth factor essential for BBB integrity; and multiple ECMs (collagens and integrins) that inhibit cell migration. Our findings suggest that via direct contact, ECs prime PCs to induce molecules to promote BBB integrity and cell survival during infection and inflammatory insult. Taken together, we provide first evidence that interaction with ECs though porous membranes induces major changes in the transcriptomic and proteomic profile of PCs. ECs influence genes involved in diverse aspects of PC function including PC maturation, cell survival, anti-viral defense, blood flow regulation, immuno-modulation and ECM deposition.

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Section: Resultsmentioning

confidence: 99%

Transcriptomic Analysis of Human Brain -Microvascular Endothelial Cell Driven Changes in -Vascular Pericytes

Kurmann

Okoniewski

Dubey

2021

Cells

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“…In the past, the oversimplified and non-physiological model obtained by simply combining PCs and ECs as co-cultures has been used [ 76 , 77 , 78 ]. Our finding of a reduced barrier function in this direct PC-EC co-culture is a clear indication for the non-physiologic nature of this model.…”

Section: Discussionmentioning

confidence: 99%

Transcryptomic Analysis of Human Brain-Microvascular Endothelial Response to -Pericytes: Cell Orientation Defines Barrier Function

Kurmann

Okoniewski

Ogunshola

et al. 2021

Cells

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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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“…However, a study by Guimarães-Camboa et al raised an important caution when interpreting pericyte differentiation and identity in experiments involving cell manipulations ex vivo [43]. This message is reinforced by data indicating that initiation and maintenance of the vascular pericyte identity in a variety of contexts may be tightly linked to a sustained physical association with the microvascular endothelium [44].…”

Section: Pericyte Differentiationmentioning

confidence: 99%

Pericytes in Vascular Development

Payne

Hoque

Houk

et al. 2020

Curr. Tissue Microenviron. Rep.

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Purpose of Review Pericytes are essential components of capillaries in many tissues and organs, contributing to vessel stability and integrity, with additional contributions to microvascular function still being discovered. We review current and foundational studies identifying pericyte differentiation mechanics and their roles in the earliest stages of vessel formation. Recent Findings Recent advances in pericyte-focused tools and models have illuminated critical aspects of pericyte biology including their roles in vascular development. Pericytes likely collaborate with endothelial cells undergoing vasculogenesis, initiating direct interactions during sprouting and intussusceptive angiogenesis. Pericytes also provide important regulation of vascular growth including mechanisms underlying vessel pruning, rarefaction, and subsequent regrowth. Summary A phenotypic transition likely occurs as pericytes shift from roles in vascular development to supporting vessel maturation, homeostasis, and physiology. We provide a forward-looking perspective on pericyte-focused studies of vascular development and applications aimed at basic and clinically relevant insights into pericyte biology.

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Transcriptome analysis reveals microvascular endothelial cell-dependent pericyte differentiation

Cited by 25 publications

References 54 publications

Transcriptomic Analysis of Human Brain -Microvascular Endothelial Cell Driven Changes in -Vascular Pericytes

Transcriptomic Analysis of Human Brain -Microvascular Endothelial Cell Driven Changes in -Vascular Pericytes

Transcryptomic Analysis of Human Brain-Microvascular Endothelial Response to -Pericytes: Cell Orientation Defines Barrier Function

Pericytes in Vascular Development

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