Single cell trajectories reveal a developmental sequence from angiogenic capillary progenitors to mature high endothelial cells

Complex interactions between stem cells, vascular cells and fibroblasts represent the substrate of building microenvironment-embedded 3D structures that can be grafted or added to bone substitute scaffolds in tissue engineering or clinical bone repair. Human Adipose-derived Stem Cells (hASCs), human umbilical vein endothelial cells (HUVECs) and normal dermal human fibroblasts (NDHF) can be mixed together in three dimensional scaffold free constructs and their behaviour will emphasize their potential use as seeding points in bone tissue engineering. Various combinations of the aforementioned cell lines were compared to single cell line culture in terms of size, viability and cell proliferation. At 5 weeks, viability dropped for single cell line spheroids while addition of NDHF to hASC maintained the viability at the same level at 5 weeks Fibroblasts addition to the 3D construct of stem cells and endothelial cells improves viability and reduces proliferation as a marker of cell differentiation toward osteogenic line.

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Single-cell RNA-seq reveals lineage-specific regulatory changes of fibroblasts and vascular endothelial cells in keloids

Journal of Investigative Dermatology ◽

10.1016/j.jid.2021.06.010 ◽

2021 ◽

Author(s):

Xuanyu Liu ◽

Wen Chen ◽

Qingyi Zeng ◽

Baihui Ma ◽

Zhujun Li ◽

...

Keyword(s):

Endothelial Cells ◽

Single Cell ◽

Vascular Endothelial Cells ◽

Vascular Endothelial ◽

Rna Seq ◽

Regulatory Changes

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Single-Cell Transcriptional Heterogeneity of Lymphatic Endothelial Cells in Normal and Inflamed Murine Lymph Nodes

Cells ◽

10.3390/cells10061371 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1371

Author(s):

Eliane Sibler ◽

Yuliang He ◽

Luca Ducoli ◽

Nadja Keller ◽

Noriki Fujimoto ◽

...

Keyword(s):

Endothelial Cells ◽

Lymph Nodes ◽

Single Cell ◽

In Silico Analysis ◽

Skin Inflammation ◽

Lymphatic Endothelial Cells ◽

Depth Analysis ◽

Subcapsular Sinus ◽

Transcriptional Changes ◽

Sinus Floor

The lymphatic system plays a crucial role in immunity and lymph nodes (LNs) undergo drastic remodeling during inflammation. Here, we used single-cell RNA sequencing to investigate transcriptional changes in lymphatic endothelial cells (LECs) in LNs draining naïve and inflamed skin. We found that subsets of LECs lining the different LN sinuses responded individually to skin inflammation, suggesting that they exert distinct functions under pathological conditions. Among the genes dysregulated during inflammation, we confirmed an up-regulation of CD200 in the LECs lining the subcapsular sinus floor with a possible function in immune regulation. Furthermore, by in silico analysis, we predicted numerous possible interactions of LECs with diverse immune cells in the LNs and found similarities in the transcriptional changes of LN LECs in different skin inflammation settings. In summary, we provide an in-depth analysis of the transcriptional landscape of LN LECs in the naïve state and in skin inflammation.

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Single cell sequencing reveals endothelial plasticity with transient mesenchymal activation after myocardial infarction

Nature Communications ◽

10.1038/s41467-021-20905-1 ◽

2021 ◽

Vol 12 (1) ◽

Cited By ~ 2

Author(s):

Lukas S. Tombor ◽

David John ◽

Simone F. Glaser ◽

Guillermo Luxán ◽

Elvira Forte ◽

...

Keyword(s):

Myocardial Infarction ◽

Endothelial Cells ◽

Endothelial Cell ◽

Single Cell ◽

Rna Sequencing ◽

Endothelial Cell Migration ◽

Metabolic Adaptation ◽

Endothelial Cell Proliferation ◽

Mesenchymal Transition ◽

Single Cell Rna Sequencing

AbstractEndothelial cells play a critical role in the adaptation of tissues to injury. Tissue ischemia induced by infarction leads to profound changes in endothelial cell functions and can induce transition to a mesenchymal state. Here we explore the kinetics and individual cellular responses of endothelial cells after myocardial infarction by using single cell RNA sequencing. This study demonstrates a time dependent switch in endothelial cell proliferation and inflammation associated with transient changes in metabolic gene signatures. Trajectory analysis reveals that the majority of endothelial cells 3 to 7 days after myocardial infarction acquire a transient state, characterized by mesenchymal gene expression, which returns to baseline 14 days after injury. Lineage tracing, using the Cdh5-CreERT2;mT/mG mice followed by single cell RNA sequencing, confirms the transient mesenchymal transition and reveals additional hypoxic and inflammatory signatures of endothelial cells during early and late states after injury. These data suggest that endothelial cells undergo a transient mes-enchymal activation concomitant with a metabolic adaptation within the first days after myocardial infarction but do not acquire a long-term mesenchymal fate. This mesenchymal activation may facilitate endothelial cell migration and clonal expansion to regenerate the vascular network.

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Single-cell metabolic imaging reveals a SLC2A3-dependent glycolytic burst in motile endothelial cells

Nature Metabolism ◽

10.1038/s42255-021-00390-y ◽

2021 ◽

Vol 3 (5) ◽

pp. 714-727

Author(s):

David Wu ◽

Devin L. Harrison ◽

Teodora Szasz ◽

Chih-Fan Yeh ◽

Tzu-Pin Shentu ◽

...

Keyword(s):

Endothelial Cells ◽

Single Cell ◽

Metabolic Imaging

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There are no A1 and A2 astrocytes in brain

10.21203/rs.3.rs-737027/v1 ◽

2021 ◽

Author(s):

Shuang-qi Gao

Keyword(s):

Endothelial Cells ◽

Single Cell ◽

Rna Sequencing ◽

Cell Sorting ◽

Reactive Astrocytes ◽

Marker Genes ◽

Set Up ◽

The Brain

Abstract Objectives The subsets of astrocytes in the brain have not been fully elucidated. Using bulk RNA sequencing, reactive astrocytes were divided into A1 versus A2. However, using single-cell RNAseq (ScRNAseq), astrocytes were divided into over two subsets. Our aim was to set up the correspondence between the fluorescent-activated cell sorting (FACS)-bulk RNAseq and ScRNAseq data. Results We found that most of reactive astrocytes (RAs) marker genes were expressed in endothelial cells but not in astrocytes, suggesting those marker genes are not suitable for astrocytic activation. The absence of A1 and A2 astrocytes in the brain.

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switchde: inference of switch-like differential expression along single-cell trajectories

Bioinformatics ◽

10.1093/bioinformatics/btw798 ◽

2016 ◽

pp. btw798 ◽

Cited By ~ 3

Author(s):

Kieran R. Campbell ◽

Christopher Yau

Keyword(s):

Single Cell ◽

Differential Expression ◽

Cell Trajectories

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Modeling cellular crosstalk and organotypic vasculature development with human iPSC-derived endothelial cells and cardiomyocytes

10.1101/2020.05.04.075846 ◽

2020 ◽

Author(s):

Emmi Helle ◽

Minna Ampuja ◽

Alexandra Dainis ◽

Laura Antola ◽

Elina Temmes ◽

...

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Endothelial Cell ◽

Single Cell ◽

Rna Sequencing ◽

Tissue Growth ◽

Cell Phenotype ◽

Cell Populations ◽

Pump System ◽

Single Cell Rna Sequencing

AbstractRationaleCell-cell interactions are crucial for the development and function of the organs. Endothelial cells act as essential regulators of tissue growth and regeneration. In the heart, endothelial cells engage in delicate bidirectional communication with cardiomyocytes. The mechanisms and mediators of this crosstalk are still poorly known. Furthermore, endothelial cells in vivo are exposed to blood flow and their phenotype is greatly affected by shear stress.ObjectiveWe aimed to elucidate how cardiomyocytes regulate the development of organotypic phenotype in endothelial cells. In addition, the effects of flow-induced shear stress on endothelial cell phenotype were studied.Methods and resultsHuman induced pluripotent stem cell (hiPSC) -derived cardiomyocytes and endothelial cells were grown either as a monoculture or as a coculture. hiPS-endothelial cells were exposed to flow using the Ibidi-pump system. Single-cell RNA sequencing was performed to define cell populations and to uncover the effects on their transcriptomic phenotypes. The hiPS-cardiomyocyte differentiation resulted in two distinct populations; atrial and ventricular. Coculture had a more pronounced effect on hiPS-endothelial cells compared to hiPS-cardiomyocytes. Coculture increased hiPS-endothelial cell expression of transcripts related to vascular development and maturation, cardiac development, and the expression of cardiac endothelial cell -specific genes. Exposure to flow significantly reprogrammed the hiPS-endothelial cell transcriptome, and surprisingly, promoted the appearance of both venous and arterial clusters.ConclusionsSingle-cell RNA sequencing revealed distinct atrial and ventricular cell populations in hiPS-cardiomyocytes, and arterial and venous-like cell populations in flow exposed hiPS-endothelial cells. hiPS-endothelial cells acquired cardiac endothelial cell identity in coculture. Our study demonstrated that hiPS-cardiomoycytes and hiPS-endothelial cells readily adapt to coculture and flow in a consistent and relevant manner, indicating that the methods used represent improved physiological cell culturing conditions that potentially are more relevant in disease modelling. In addition, novel cardiomyocyte-endothelial cell crosstalk mediators were revealed.

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Single-cell transcriptome analysis reveals cellular heterogeneity in the ascending aortas of normal and high-fat diet-fed mice

Experimental & Molecular Medicine ◽

10.1038/s12276-021-00671-2 ◽

2021 ◽

Vol 53 (9) ◽

pp. 1379-1389

Author(s):

Hao Kan ◽

Ka Zhang ◽

Aiqin Mao ◽

Li Geng ◽

Mengru Gao ◽

...

Keyword(s):

Endothelial Cells ◽

Single Cell ◽

High Fat Diet ◽

Cell Types ◽

Ascending Aorta ◽

Normal Diet ◽

Cellular Heterogeneity ◽

Cellular Composition ◽

High Fat ◽

Aortic Diseases

AbstractThe aorta contains numerous cell types that contribute to vascular inflammation and thus the progression of aortic diseases. However, the heterogeneity and cellular composition of the ascending aorta in the setting of a high-fat diet (HFD) have not been fully assessed. We performed single-cell RNA sequencing on ascending aortas from mice fed a normal diet and mice fed a HFD. Unsupervised cluster analysis of the transcriptional profiles from 24,001 aortic cells identified 27 clusters representing 10 cell types: endothelial cells (ECs), fibroblasts, vascular smooth muscle cells (SMCs), immune cells (B cells, T cells, macrophages, and dendritic cells), mesothelial cells, pericytes, and neural cells. After HFD intake, subpopulations of endothelial cells with lipid transport and angiogenesis capacity and extensive expression of contractile genes were defined. In the HFD group, three major SMC subpopulations showed increased expression of extracellular matrix-degradation genes, and a synthetic SMC subcluster was proportionally increased. This increase was accompanied by upregulation of proinflammatory genes. Under HFD conditions, aortic-resident macrophage numbers were increased, and blood-derived macrophages showed the strongest expression of proinflammatory cytokines. Our study elucidates the nature and range of the cellular composition of the ascending aorta and increases understanding of the development and progression of aortic inflammatory disease.

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Integrated Single Cell Atlas of Endothelial Cells of the Human Lung

Circulation ◽

10.1161/circulationaha.120.052318 ◽

2021 ◽

Author(s):

Jonas C. Schupp ◽

Taylor S. Adams ◽

Carlos Cosme Jr. ◽

Micha Sam Brickman Raredon ◽

Yifan Yuan ◽

...

Keyword(s):

Endothelial Cells ◽

Pulmonary Hypertension ◽

Single Cell ◽

Differential Expression ◽

Human Lung ◽

Differential Expression Analysis ◽

Cell Types ◽

Marker Genes ◽

Lung Endothelium ◽

Lung Endothelial Cells

Background: The cellular diversity of the lung endothelium has not been systematically characterized in humans. Here, we provide a reference atlas of human lung endothelial cells (ECs) to facilitate a better understanding of the phenotypic diversity and composition of cells comprising the lung endothelium. Methods: We reprocessed human control single cell RNA sequencing (scRNAseq) data from six datasets. EC populations were characterized through iterative clustering with subsequent differential expression analysis. Marker genes were validated by fluorescent microscopy and in situ hybridization. scRNAseq of primary lung ECs cultured in-vitro was performed. The signaling network between different lung cell types was studied. For cross species analysis or disease relevance, we applied the same methods to scRNAseq data obtained from mouse lungs or from human lungs with pulmonary hypertension. Results: Six lung scRNAseq datasets were reanalyzed and annotated to identify over 15,000 vascular EC cells from 73 individuals. Differential expression analysis of EC revealed signatures corresponding to endothelial lineage, including pan-endothelial, pan-vascular and subpopulation-specific marker gene sets. Beyond the broad cellular categories of lymphatic, capillary, arterial and venous ECs, we found previously indistinguishable subpopulations: among venous EC, we identified two previously indistinguishable populations, pulmonary-venous ECs (COL15A1neg) localized to the lung parenchyma and systemic-venous ECs (COL15A1pos) localized to the airways and the visceral pleura; among capillary EC, we confirmed their subclassification into recently discovered aerocytes characterized by EDNRB, SOSTDC1 and TBX2 and general capillary EC. We confirmed that all six endothelial cell types, including the systemic-venous EC and aerocytes, are present in mice and identified endothelial marker genes conserved in humans and mice. Ligand-receptor connectome analysis revealed important homeostatic crosstalk of EC with other lung resident cell types. scRNAseq of commercially available primary lung ECs demonstrated a loss of their native lung phenotype in culture. scRNAseq revealed that the endothelial diversity is maintained in pulmonary hypertension. Our manuscript is accompanied by an online data mining tool (www.LungEndothelialCellAtlas.com). Conclusions: Our integrated analysis provides the comprehensive and well-crafted reference atlas of lung endothelial cells in the normal lung and confirms and describes in detail previously unrecognized endothelial populations across a large number of humans and mice.

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