Imbalanced Activation of Wnt-/β-Catenin-Signaling in Liver Endothelium Alters Normal Sinusoidal Differentiation

Endothelial wingless-related integration site (Wnt)-/β-catenin signaling is a key regulator of the tightly sealed blood–brain barrier. In the hepatic vascular niche angiokine-mediated Wnt signaling was recently identified as an important regulator of hepatocyte function, including the determination of final adult liver size, liver regeneration, and metabolic liver zonation. Within the hepatic vasculature, the liver sinusoidal endothelial cells (LSECs) are morphologically unique and functionally specialized microvascular endothelial cells (ECs). Pathological changes of LSECs are involved in chronic liver diseases, hepatocarcinogenesis, and liver metastasis. To comprehensively analyze the effects of endothelial Wnt-/β-catenin signaling in the liver, we used endothelial subtype-specific Clec4g-iCre mice to generate hepatic ECs with overexpression of Ctnnb1. In the resultant Clec4g-iCretg/wt;Ctnnb1(Ex3)fl/wt (Ctnnb1OE−EC) mice, activation of endothelial Wnt-/β-catenin signaling resulted in sinusoidal transdifferentiation with disturbed endothelial zonation, that is, loss of midzonal LSEC marker lymphatic vessel endothelial hyaluronic acid receptor 1 (Lyve1) and enrichment of continuous EC genes, such as cluster of differentiation (CD)34 and Apln. Notably, gene set enrichment analysis revealed overrepresentation of brain endothelial transcripts. Activation of endothelial Wnt-/β-catenin signaling did not induce liver fibrosis or alter metabolic liver zonation, but Ctnnb1OE−EC mice exhibited significantly increased plasma triglyceride concentrations, while liver lipid content was slightly reduced. Ctnnb1 overexpression in arterial ECs of the heart has been reported previously to cause cardiomyopathy. As Clec4g-iCre is active in a subset of cardiac ECs, it was not unexpected that Ctnnb1OE−EC mice showed reduced overall survival and cardiac dysfunction. Altogether, balanced endothelial Wnt-/β-catenin signaling in the liver is required for normal LSEC differentiation and for maintenance of normal plasma triglyceride levels.

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Age-Dependent Transcriptional Alterations in Cardiac Endothelial Cells

Physiological Genomics ◽

10.1152/physiolgenomics.00037.2021 ◽

2021 ◽

Author(s):

Uchenna Emechebe ◽

Jonathan William Nelson ◽

Nabil J. Alkayed ◽

Sanjiv Kaul ◽

Andrew C Adey ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Single Cell ◽

Significant Risk ◽

Enrichment Analysis ◽

Gene Set Enrichment Analysis ◽

Mouse Heart ◽

Age Related ◽

Organ Systems ◽

Transcriptional Alterations

Aging is a significant risk factor for cardiovascular disease. Despite the fact that endothelial cells play critical roles in cardiovascular function and disease, the molecular impact of aging on this cell population in many organ systems remains unknown. In this study, we sought to determine age-associated transcriptional alterations in cardiac endothelial cells. Highly enriched populations of endothelial cells (ECs) isolated from the heart, brain and kidney of young (3 months) and aged (24 months) C57/BL6 mice were profiled for RNA expression via bulk RNA sequencing. Approximately 700 cardiac endothelial transcripts significantly differ by age. Gene set enrichment analysis indicated similar patterns for cellular pathway perturbations. Receptor-ligand comparisons indicated parallel alterations in age-affected circulating factors and cardiac endothelial-expressed receptors. Single-cell RNA-seq analysis identified 9 distinct endothelial cell subtypes in the heart with an age-associated population shift observed for the Aplnr-enriched endothelial cell clusters. Gene and pathway enrichment analyses show that age-related transcriptional response of cardiac endothelial cells is distinct from that of endothelial cells derived from the brain or kidney vascular bed. Furthermore, single-cell analysis identified 9 distinct EC subtypes, and shows that the Aplnr-enriched subtype is reduced with age in mouse heart. Finally, we identify age-dysregulated genes in specific aged cardiac endothelial subtypes.

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Circular RNA circ_0003204 inhibits proliferation, migration and tube formation of endothelial cell in atherosclerosis via miR-370-3p/TGFβR2/phosph-SMAD3 axis

Journal of Biomedical Science ◽

10.1186/s12929-019-0595-9 ◽

2020 ◽

Vol 27 (1) ◽

Cited By ~ 19

Author(s):

Shanchao Zhang ◽

Guixiang Song ◽

Jing Yuan ◽

Shan Qiao ◽

Shan Xu ◽

...

Keyword(s):

Endothelial Cells ◽

Ectopic Expression ◽

Enrichment Analysis ◽

Tube Formation ◽

Gene Set Enrichment Analysis ◽

Luciferase Assay ◽

Circular Rnas ◽

Human Aorta ◽

Cerebral Atherosclerosis ◽

Potential Biomarker

Abstract Background Circular RNAs (circRNAs) represent a class of non-coding RNAs (ncRNAs) which are widely expressed in mammals and tissue-specific, of which some could act as critical regulators in the atherogenesis of cerebrovascular disease. However, the underlying mechanisms by which circRNA regulates the ectopic phenotype of endothelial cells (ECs) in atherosclerosis remain largely elusive. Methods CCK-8, transwell, wound healing and Matrigel assays were used to assess cell viability, migration and tube formation. QRT-qPCR and Immunoblotting were used to examine targeted gene expression in different groups. The binding sites of miR-370-3p (miR-370) with TGFβR2 or hsa_circ_0003204 (circ_0003204) were predicted using a series of bioinformatic tools, and validated using dual luciferase assay and RNA immunoprecipitation (RIP) assay. The localization of circ_0003204 and miR-370 in ECs were investigated by fluorescence in situ hybridization (FISH). Gene function and pathways were enriched through Metascape and gene set enrichment analysis (GSEA). The association of circ_0003204 and miR-370 in extracellular vesicles (EVs) with clinical characteristics of patients were investigated using multiple statistical analysis. Results Circ_0003204, mainly located in the cytoplasm of human aorta endothelial cells (HAECs), was upregulated in the ox-LDL-induced HAECs. Functionally, the ectopic expression of circ_0003204 inhibited proliferation, migration and tube formation of HAECs exposed to ox-LDL. Mechanically, circ_0003204 could promote protein expression of TGFβR2 and its downstream phosph-SMAD3 through sponging miR-370, and miR-370 targeted the 3′ untranslated region (UTR) of TGFβR2. Furthermore, the expression of circ_0003204 in plasma EVs was upregulated in the patients with cerebral atherosclerosis, and represented a potential biomarker for diangnosis and prognosis of cerebrovascular atherogenesis. Conclusions Circ_0003204 could act as a novel stimulator for ectopic endothelial inactivation in atherosclerosis and a potential biomarker for cerebral atherosclerosis.

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Identification of Key Candidate Genes and Pathways of Candida albicans-Infected Human Umbilical Vein Endothelial Cells and Drug Screening

Indian Journal of Microbiology ◽

10.1007/s12088-019-00847-5 ◽

2019 ◽

Vol 60 (1) ◽

pp. 62-69

Author(s):

Wei Jiang ◽

Ping Liu ◽

Jianlei Zhang ◽

Wenjie Yang

Keyword(s):

Endothelial Cells ◽

Candida Albicans ◽

Signaling Pathways ◽

Umbilical Vein ◽

Enrichment Analysis ◽

Cell Receptor ◽

Gene Set Enrichment Analysis ◽

Purine Nucleotide ◽

Human Umbilical Vein ◽

Umbilical Vein Endothelial Cells

AbstractCandida albicans is a common opportunistic pathogen that can cause serious infection by blood transmission. C. albicans enters the blood circulation and adheres to the endothelial cells of the vascular wall. However, the detailed mechanism of the effect of C. albicans on the endothelial cells remains unclear. In this study, the microarray expression profile of human umbilical vein endothelial cells exposed to C. albicans was analyzed. The 191 up-regulated genes were enriched in TNF, T cell receptor, and NF-kappa B signaling pathways. The 71 down-regulated genes were enriched in pyruvate metabolic, purine nucleotide metabolic, purine nucleotide biosynthetic, and humoral immune response processes. Gene set enrichment analysis showed that apoptosis, oxidative phosphorylation, IL6/JAK/STAT3 signaling pathways were enriched. Moreover, two hub genes with a high degree of connectivity, namely, MYC and IL6, were selected. Molecular screening of traditional Chinese medicine libraries was performed on the basis of the structure of MYC protein. The okanin had the highest docking score. MYC might be used as molecular targets for treatment. In addition, okanin may inhibit the infection of C. albicans. Thus, MYC can be subjected to further research.

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Integrated Transcriptomic and Translatomic Inquiry of the Role of Betaine on Lipid Metabolic Dysregulation Induced by a High-Fat Diet

Frontiers in Nutrition ◽

10.3389/fnut.2021.751436 ◽

2021 ◽

Vol 8 ◽

Author(s):

Tengda Huang ◽

Lin Yu ◽

Hongyuan Pan ◽

Zeqiang Ma ◽

Tian Wu ◽

...

Keyword(s):

Fatty Acid ◽

High Fat Diet ◽

Liver Lipid ◽

Liver Steatosis ◽

Enrichment Analysis ◽

Gene Set Enrichment Analysis ◽

Translational Efficiency ◽

High Fat ◽

Alcoholic Fatty Liver ◽

Translational Level

An excessive high-fat/energy diet is a major cause of obesity and linked complications, such as non-alcoholic fatty liver disease (NAFLD). Betaine has been shown to effectively improve hepatic lipid metabolism. However, the mechanistic basis for this improvement is largely unknown. Herein, integration of mRNA sequencing and ribosome footprints profiling (Ribo-seq) was used to investigate the means by which betaine alleviates liver lipid metabolic disorders induced by a high-fat diet. For the transcriptome, gene set enrichment analysis demonstrated betaine to reduce liver steatosis by up-regulation of fatty acid beta oxidation, lipid oxidation, and fatty acid catabolic processes. For the translatome, 574 differentially expressed genes were identified, 17 of which were associated with the NAFLD pathway. By combined analysis of transcriptome and translatome, we found that betaine had the greater effect on NAFLD at the translational level. Further, betaine decreased translational efficiency (TE) for IDI1, CYP51A1, TM7SF2, and APOA4, which are related to lipid biosynthesis. In summary, this study demonstrated betaine alleviating lipid metabolic dysfunction at the translational level. The transcriptome and translatome data integration approach used herein provides for a new understanding of the means by which to treat NAFLD.

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Angiocrine signaling in the hepatic sinusoids in health and disease

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00118.2016 ◽

2016 ◽

Vol 311 (2) ◽

pp. G246-G251 ◽

Cited By ~ 16

Author(s):

Enis Kostallari ◽

Vijay H. Shah

Keyword(s):

Endothelial Cells ◽

Cell Activation ◽

Stellate Cell ◽

Cell Types ◽

Hepatocyte Proliferation ◽

Liver Sinusoidal Endothelial Cells ◽

Functional Changes ◽

Liver Zonation ◽

Health And Disease ◽

Short Synthesis

The capillary network irrigating the liver is important not only for nutrient and oxygen delivery, but also for the signals distributed to other hepatic cell types necessary to maintain liver homeostasis. During development, endothelial cells are a key component in liver zonation. In adulthood, they maintain hepatic stellate cells and hepatocytes in quiescence. Their importance in pathobiology is highlighted in liver regeneration and chronic liver diseases, where they coordinate paracrine cell behavior. During regeneration, liver sinusoidal endothelial cells induce hepatocyte proliferation and angiogenesis. During fibrogenesis, they undergo morphological and functional changes, which are reflected by their role in hepatic stellate cell activation, inflammation, and distorted sinusoidal structure. Therapeutic strategies to target angiocrine signaling are in progress but are in the early stages. Here, we offer a short synthesis of recent studies on angiocrine signaling in liver homeostasis, regeneration, and fibrogenesis.

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ELTD1 Activation Induces an Endothelial-EMT Transition to a Myofibroblast Phenotype

International Journal of Molecular Sciences ◽

10.3390/ijms222011293 ◽

2021 ◽

Vol 22 (20) ◽

pp. 11293

Author(s):

Helen Sheldon ◽

John Alexander ◽

Esther Bridges ◽

Lucia Moreira ◽

Svetlana Reilly ◽

...

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Wound Repair ◽

Network Formation ◽

Smooth Muscle Actin ◽

Focal Adhesions ◽

Enrichment Analysis ◽

Gene Set Enrichment Analysis ◽

Immune Response Gene ◽

Cell Contact

ELTD1 is expressed in endothelial and vascular smooth muscle cells and has a role in angiogenesis. It has been classified as an adhesion GPCR, but as yet, no ligand has been identified and its function remains unknown. To establish its role, ELTD1 was overexpressed in endothelial cells. Expression and consequently ligand independent activation of ELTD1 results in endothelial-mesenchymal transistion (EndMT) with a loss of cell-cell contact, formation of stress fibres and mature focal adhesions and an increased expression of smooth muscle actin. The effect was pro-angiogenic, increasing Matrigel network formation and endothelial sprouting. RNA-Seq analysis after the cells had undergone EndMT revealed large increases in chemokines and cytokines involved in regulating immune response. Gene set enrichment analysis of the data identified a number of pathways involved in myofibroblast biology suggesting that the endothelial cells had undergone a type II EMT. This type of EMT is involved in wound repair and is closely associated with inflammation implicating ELTD1 in these processes.

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Serum Calcium is Related to the Degree of Artery Stenosis in Acute Ischemic Stroke

Cellular Physiology and Biochemistry ◽

10.1159/000489069 ◽

2018 ◽

Vol 46 (3) ◽

pp. 1189-1197

Author(s):

Jiayan Wu ◽

Junchao Xie ◽

Yanxin Zhao ◽

Li Gong ◽

Xueyuan Liu ◽

...

Keyword(s):

Gene Expression ◽

Endothelial Cells ◽

Ischemic Stroke ◽

Acute Ischemic Stroke ◽

Serum Calcium ◽

Density Lipoprotein ◽

Enrichment Analysis ◽

Artery Stenosis ◽

Gene Set Enrichment Analysis ◽

Signaling System

Background/Aims: Acute ischemic stroke is caused by stenosis of artery supplying to brain. We aimed to detect some metabolites in the serum that would be related to the degree of artery stenosis and to analyze potential mechanisms. Methods: Patients diagnosed with acute ischemic stroke were divided into two groups according to their degree of artery stenosis (which was determined by computed tomographic angiography): a mild group (stenosis ≤ 30%) and a severe group (stenosis > 30%). Serum from these patients was collected, and we focused on the differences in the concentrations of calcium, uric acid, low density lipoprotein and homocysteine. The dataset GSE11583 from the Gene Expression Omnibus database was analyzed to find the potential mechanism using bioinformatics methods. Results: Among the four metabolites, the only difference that reached significance between the two groups was in the concentration of calcium in serum (2.27±0.08 mmol/L vs 2.21±0.08 mmol/L). By comparing the gene expression levels between normal endothelial cells and adaptive remodeling endothelial cells in GSE11583, we identified 51 upregulated and 40 downregulated genes in adaptive remodeling endothelial cells. The gene set enrichment analysis revealed that upregulated genes were enriched in a phosphatidylinositol signaling system, which is closely involved in the calcium signaling pathway. Conclusion: Our results suggest that the concentration of serum calcium is higher in patients with more severe artery stenosis lesions and that the phosphatidylinositol signaling system is a key biological pathway involved in this process.

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Developmental programming in human umbilical cord vein endothelial cells following fetal growth restriction

Clinical Epigenetics ◽

10.1186/s13148-020-00980-9 ◽

2020 ◽

Vol 12 (1) ◽

Author(s):

Fieke Terstappen ◽

Jorg J. A. Calis ◽

Nina D. Paauw ◽

Jaap A. Joles ◽

Bas B. van Rijn ◽

...

Keyword(s):

Dna Methylation ◽

Endothelial Cells ◽

Rna Sequencing ◽

Fetal Growth ◽

Fetal Growth Restriction ◽

Enrichment Analysis ◽

Growth Restriction ◽

Gene Set Enrichment Analysis ◽

Gene Set Enrichment ◽

Gene Sets

Abstract Background Fetal growth restriction (FGR) is associated with an increased susceptibility for various noncommunicable diseases in adulthood, including cardiovascular and renal disease. During FGR, reduced uteroplacental blood flow, oxygen and nutrient supply to the fetus are hypothesized to detrimentally influence cardiovascular and renal programming. This study examined whether developmental programming profiles, especially related to the cardiovascular and renal system, differ in human umbilical vein endothelial cells (HUVECs) collected from pregnancies complicated by placental insufficiency-induced FGR compared to normal growth pregnancies. Our approach, involving transcriptomic profiling by RNA-sequencing and gene set enrichment analysis focused on cardiovascular and renal gene sets and targeted DNA methylation assays, contributes to the identification of targets underlying long-term cardiovascular and renal diseases. Results Gene set enrichment analysis showed several downregulated gene sets, most of them involved in immune or inflammatory pathways or cell cycle pathways. seven of the 22 significantly upregulated gene sets related to kidney development and four gene sets involved with cardiovascular health and function were downregulated in FGR (n = 11) versus control (n = 8). Transcriptomic profiling by RNA-sequencing revealed downregulated expression of LGALS1, FPR3 and NRM and upregulation of lincRNA RP5-855F14.1 in FGR compared to controls. DNA methylation was similar for LGALS1 between study groups, but relative hypomethylation of FPR3 and hypermethylation of NRM were present in FGR, especially in male offspring. Absolute differences in methylation were, however, small. Conclusion This study showed upregulation of gene sets related to renal development in HUVECs collected from pregnancies complicated by FGR compared to control donors. The differentially expressed gene sets related to cardiovascular function and health might be in line with the downregulated expression of NRM and upregulated expression of lincRNA RP5-855F14.1 in FGR samples; NRM is involved in cardiac remodeling, and lincRNAs are correlated with cardiovascular diseases. Future studies should elucidate whether the downregulated LGALS1 and FPR3 expressions in FGR are angiogenesis-modulating regulators leading to placental insufficiency-induced FGR or whether the expression of these genes can be used as a biomarker for increased cardiovascular risk. Altered DNA methylation might partly underlie FPR3 and NRM differential gene expression differences in a sex-dependent manner.

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ELTD1 Activation Induces an Endothelial-EMT Transition to a Myofibroblast Phenotype

10.20944/preprints202109.0402.v1 ◽

2021 ◽

Author(s):

Helen Sheldon ◽

John Alexander ◽

Esther M. Bridges ◽

Lucia Moreira ◽

Svetlana Reilly ◽

...

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Wound Repair ◽

Network Formation ◽

Smooth Muscle Actin ◽

Focal Adhesions ◽

Enrichment Analysis ◽

Gene Set Enrichment Analysis ◽

Immune Response Gene ◽

Cell Contact

ELTD1 is expressed in endothelial and vascular smooth muscle cells and has a role in angiogenesis. It has been classified as an adhesion GPCR, but as yet, no ligand has been identified and its function remains unknown. To establish its role, ELTD1 was overexpressed in endothelial cells. Expression and consequently ligand independent activation of ELTD1 results in EndMT with a loss of cell-cell contact, formation of stress fibres and mature focal adhesions and an increased expression of smooth muscle actin. The effect was pro-angiogenic, increasing Matrigel network formation and endothelial sprouting. RNA-Seq analysis after the cells had undergone EndMT revealed large increases in chemokines and cytokines involved in regulating immune response. Gene set enrichment analysis of the data identified a number of pathways involved in myofibroblast biology suggesting that the endothelial cells had undergone a type II EMT. This type of EMT is involved in wound repair and is closely associated with inflammation implicating ELTD1 in these processes.

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