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Analysis of extracellular vesicle microRNA profiles reveals distinct blood and lymphatic endothelial cell origins [mRNA]

ABSTRACT: Extracellular Vesicles (EVs) are crucial mediators of cell-to-cell communication in physiology but also in pathological conditions. Specifically, EVs released from the vasculature into blood were found to be quantitatively and qualitatively different in diseases versus healthy states. However, our understanding on EVs derived from the lymphatic system is still scarce. In this study we compared the mRNA and miRNA expression in blood vascular (BEC) and lymphatic (LEC) endothelial cells. After characterization of the EVs by fluo-triggered flow cytometry, nanoparticle tracking analysis and cryo-EM we utilized small RNA-sequencing to characterize miRNA signatures in the EVs and thereof identify cell-type specific miRNAs in BEC and LEC. We believe that our data provide a solid basis for further functional in vitro and in vivo studies addressing the role of EVs in the blood and lymphatic vasculature.

ORGANISM(S): Homo sapiens

PROVIDER: GSE232377 | GEO | 2024/01/17

REPOSITORIES: GEO

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Project description:The lymphatic vascular system plays important roles in the maintenance of interstitial fluid pressure, the afferent immune response and the absorption of dietary lipids. However, the molecular mechanisms that control lymphatic vessel network maturation and function remain largely unknown. To identify novel players in lymphatic vessel function, we isolated pure populations of lymphatic and blood vascular endothelial cells from mouse intestine using fluorescence-activated high-speed cell sorting and performed transcriptional profiling. We found that the axonal guidance molecules semaphorin 3A (Sema3A) and Sema3D were specifically expressed by lymphatic vessels. Quantitative PCR of ex vivo isolated cells and immunohistochemical analysis confirmed these results. Importantly, we found that the semaphorin receptor neuropilin-1 (Nrp-1) is expressed on the valves of collecting lymphatic vessels. Treatment of mice in utero (E12.5-E16.5) with an antibody that blocks Sema3A binding to Nrp-1, but not with an antibody that blocks VEGFA binding to Nrp-1, resulted in abnormal development of collecting lymphatic vessels and valves, and aberrant smooth muscle cell coverage. Conversely, Sema3A-deficient mice displayed branching defects of collecting lymphatic vessels as well as impaired valve development. Together, these results reveal an unanticipated role of Sema3A/Nrp-1 signaling in the maturation of the lymphatic vascular network. Colon single-cell suspensions were prepared by a fast protocol that minimizes the RNA degradation. Fluorescence-activated cell sorting (FACS) was used to sort blood vascular endothelial cells (BEC) and lymphatic endothelial cells (LEC). 4 animal-matched pairs of LEC and BEC were chosen based on the quality of extracted and amplified material to provide homogenous groups of biological replicates. This gave 8 samples to analyze. Samples present LEC and BEC isolated from 4 healthy normal mice. The 4 mice used present the 4 biological replicates.

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Analysis of extracellular vesicle microRNA profiles reveals distinct blood and lymphatic endothelial cell origins [mRNA]

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