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Glycan analysis of promary human lymphatic endothlial cells (LEX) in their resting and activated states, in normal and tumor tissues

ABSTRACT: Trafficking of leukocytes (dendritic cells, memory T cells, neutrophils) and tumor cells through the lymphatic network is a key process in inflammation and immunity and an important mechanism in metastatic spread of human cancers (1-3). Such trafficking involves both communication with and passage across lymphatic endothelium, the distinct endothelium that lines lymphatic vessels within the peripheral tissues and forms the lymphatic sinuses within lymph nodes. However, in comparison with blood vascular endothelium, there is only a rudimentary understanding of the molecular phenotype of lymphatic endothelium, and only a basic knowledge of the glycoconjugates that regulate leukocyte-endothelial and tumor cell-endothelial interactions in the lymphatic compartment.

ORGANISM(S): Homo sapiens

PROVIDER: GSE29933 | GEO | 2011/06/14

SECONDARY ACCESSION(S): PRJNA140879

REPOSITORIES: GEO

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Glycan analysis of promary human lymphatic endothlial cells (LEX) in their resting and activated states, in normal and tumor tissues

Project description:Trafficking of leukocytes (dendritic cells, memory T cells, neutrophils) and tumor cells through the lymphatic network is a key process in inflammation and immunity and an important mechanism in metastatic spread of human cancers (1-3). Such trafficking involves both communication with and passage across lymphatic endothelium, the distinct endothelium that lines lymphatic vessels within the peripheral tissues and forms the lymphatic sinuses within lymph nodes. However, in comparison with blood vascular endothelium, there is only a rudimentary understanding of the molecular phenotype of lymphatic endothelium, and only a basic knowledge of the glycoconjugates that regulate leukocyte-endothelial and tumor cell-endothelial interactions in the lymphatic compartment. We would anticipate that changes in glycosylation of LEC cell surface proteins following activation might affect important functions associated withy LEC including eg. interactions with leukocytes and/or sequestration of GAG-binding chemokines. We already have shown that ligand binding to the lymphatic endothelial hyaluronan receptor LYVE-1 is reversibly masked by terminal sialation in LEC and that functional regulation of LYVE-1 is likely to be important in inflammation. An advantage of our proposal is that we can routinely isolate primary LEC in relatively large numbers, and have the necessary ethical approval to do so from human tissue. Glycan analysis of primary human lymphatic endothelial cells (LEC) in their resting and activated states, in normal and tumour tissues and a comparison of the LEC glycan structures with those from blood vessel endothelial cells

2011-06-14 | E-GEOD-29933 | biostudies-arrayexpress

Transcription profiling of human lymphatic endothelial cells, mouse dermal cells and human skin cells reveals an inflammation-induced mechanism for leukocyte transmigration of lymphatic vessel endothelium.

Project description:The exit of antigen-presenting cells (APC) and lymphocytes from inflamed skin to afferent lymph is vital for the initiation and maintenance of dermal immune responses. How such exit is achieved and how cells transmigrate the distinct endothelium of lymphatic vessels is however unknown. Here we show that inflammatory cytokines trigger activation of dermal lymphatic endothelial cells (LEC) leading to expression of the key leukocyte adhesion receptors ICAM-1, VCAM-1 and E-selectin, as well as a discrete panel of chemokines and other potential regulators of leukocyte transmigration. Furthermore, we show that both ICAM-1 and VCAM-1 are induced in the dermal lymphatic vessels of mice exposed to skin contact hypersensitivity where they mediate lymph node trafficking of DC via afferent lymphatics. Lastly, we show that TNF_-stimulates both DC adhesion and transmigration of dermal LEC monolayers in vitro and that the process is efficiently inhibited by ICAM-1 and VCAM-1 adhesion-blocking mAbs. These results reveal a CAM-mediated mechanism for recruiting leukocytes to the lymph nodes in inflammation and highlight the process of lymphatic transmigration as a potential new target for anti-inflammatory therapy. Experiment Overall Design: Global gene expression profile of normal dermal lymphatic endothelial cells cultured in media alone (no TNF) compared to that of normal dermal lymphatic endothelial cells stimulated with TNFalpha, 1 ng/ml for 48h.Triplicate biological samples were analyzed from human lymphatic endothelial cells (3 x controls; 3 x TNF treated) and a single sample analyzed from mouse lymphatic endothelial cells (1 x controls; 1 x TNF treated).

2008-11-07 | E-GEOD-6257 | biostudies-arrayexpress

Expression data from collecting lymphatic vessels

Project description:Lymphogenous metastasis is an important event in the progression of many human cancers, and is associated with expression of vascular endothelial growth factor-D (VEGF-D). Changes to the lymphatic vasculature can occur during metastasis, and may aid metastatic spread. We investigated the effect of tumour derived VEGFD on the endothelium of the collecting lymphatic vessels draining primary tumors. We used microarrays to detail the changes in gene expression in the collecting lymphatic endothelium of mice with 293EBNA xenografts compared to 293EBNA xenografts overexpressing VEGFD. Mice were injected with 293EBNA cells (transfected with either empty APEX vector, or vector containing VEGFD) and tumours were allowed to grow to size. Mice were sacrificed and collecting lymphatic vessels were dissected. The endothelial cell population was isolated and RNA was extracted and hybridized on Affymetrix microarrays.

2012-01-15 | E-GEOD-34135 | biostudies-arrayexpress

Global gene expression differences between blood- and lymphatic-specific endothelial colony forming cells

Project description:The identification of circulating endothelial progenitor cells has led to speculation regarding their origin as well as their contribution to neovascular development. Two distinct types of endothelium make up the blood and lymphatic vessel system. However, it has yet to be determined whether there are distinct lymphatic-specific circulating endothelial progenitor cells. We isolated circulating endothelial colony forming cells (ECFCs) from whole peripheral blood. These cells are endothelial in nature, as defined by their expression of endothelial markers and their ability to undergo capillary morphogenesis in three-dimensional culture. A subset of isolated colonies express markers of lymphatic endothelium, including VEGFR-3 and Prox-1, with low levels of VEGFR-1, a blood endothelial marker, while the bulk of the isolated cells express high VEGFR-1 levels with low VEGFR-3 and Prox-1 expression. The different isolates have differential responses to VEGF-C, a lymphatic endothelial specific cytokine, strongly suggesting that there are lymphatic specific and blood specific ECFCs. Global analysis of gene expression revealed key differences in the regulation of pathways involved in cellular differentiation between blood and lymphatic-specific ECFCs. These data indicate that there are two distinguishable circulating ECFC types, blood and lymphatic, which are likely to have discrete functions during neovascularization. RNA was isolated from 2 blood-specific ECFC cell lines and 2 lymphatic-specific ECFC cell lines 3 separate times each

2014-06-23 | E-GEOD-54416 | biostudies-arrayexpress

Expression profile of human lymphatic endothelial cells under static or oscillatory shear stress conditions in the presence or absence of FOXC2

Project description:Lymphatic valves are specialized units regularly distributed along collecting vessels that allow unidirectional forward propulsion of the lymph, and its efficient transport from tissues to the bloodstream. Lymphatic endothelial cells that cover lymphatic valve sinuses are subjected to complex flow patterns, due to recirculation of the lymph during the collecting vessel pumping cycle. They also express high levels of FOXC2 transcription factor. We used microarrays to study the transcriptional networks controlled by FOXC2 in human lymphatic endothelial cells subjected to oscillatory shear stress or cultured under static conditions. Human lymphatic endothelial cells were transfected with control or FOXC2 siRNAs and subjected to 24-hour oscillatory shear stress (1 dyn/cm2; 1/4 Hz) or kept under static conditions as a control. RNA were amplified and hybridized on Affymetrix Human Gene 1.0 ST Arrays. The experiment was run twice independently, using each time a different siRNA to knockdown FOXC2, as previously described (Sabine et al, 2012, Dev Cell).

2015-11-09 | E-GEOD-60152 | biostudies-arrayexpress

Expression profile of human lymphatic endothelial cells under static or oscillatory shear stress conditions in the presence or absence of FOXC2

2015-11-09 | GSE60152 | GEO

Expression data from collecting lymphatic vessels

2012-01-15 | GSE34135 | GEO

Lymphatic endothelial cell identity is controlled by a GATA2 bound enhancer [RNA-seq]

Project description:Transcriptional enhancer elements are responsible for orchestrating the temporal and spatial control over gene expression that is crucial for programming cell identity during development. Here, we describe a novel enhancer element important for regulating Prox1 expression in lymphatic endothelial cells. This evolutionarily conserved enhancer is bound by key lymphatic transcriptional regulators including GATA2, FOXC2, NFATC1 and PROX1. CRISPR-Cas9 genome editing of this enhancer element revealed that deletion of only 5 nucleotides encompassing the GATA2 binding site has a dramatic impact on lymphatic vascular development; mice homozygous for this deletion die soon after birth exhibiting profound lymphatic vascular defects. Lymphatic endothelial cells in enhancer mutant mice exhibit reduced levels of genes characteristic of lymphatic endothelial cell identity and acquire characteristics of hemogenic endothelium, including the capacity to generate hematopoietic cells. These data reveal the first transcriptional enhancer element important for regulating Prox1 expression and lymphatic endothelial cell identity and suggest that Prox1 is important for repressing hemogenic cell identity in the lymphatic endothelium.

2022-11-21 | GSE129633 | GEO

Lymphatic endothelial cell identity is controlled by a GATA2 bound enhancer [ChIP-seq]

2022-11-21 | GSE129632 | GEO

Evaluation of the effects of shear stress on human dermal microvascular blood and lymphatic endothelial cells

Project description:The lymphatic system removes fluid from the interstitial space and returns it to the blood with a tremendous capacity: during inflammation, lymph flow rates can increase dramatically; however, during chronic lymphedema, there is little or no flow. The ability of lymphatic endothelium to sense and actively regulate this function is unknown, and shear stress is likely a key indicator of lymph flow. We profiled gene expression in human dermal microvascular lymphatic endothelial cells exposed to 0, 2 and 20 dyn/cm2 shear stress as representative of chronic lymphedema, normal, and acute inflammatory conditions, respectively. We found important adaptive responses correlated to multiple aspects of lymphatic function. Importantly, shear stress upregulated intracellular water and solute transporters while decreasing cell-cell adhesion and basement membrane components and increasing cell-matrix interactions. This data indicate that during high loading conditions, both passive and active drainage function increases, while conversely when fluid drainage is blocked, transport function is diminished in the lymphatic endothelium. These data demonstrate the first functional-adaptive response of lymphatic endothelium to flow conditions, thus indicating that the lymphatic endothelium plays an active role in regulating their function. Keywords: Shear stress, dose response, cell type comparison Lymphatic endothelial cells were subjected to 0, 2, or 20 dyn/cm2 shear stress; blood endothelial cells were subjected to 0 or 20 dyn/cm2 shear stress. Four samples were used for each cell type/shear level group for a total of 20 samples. Each sample was independently compared to human universal reference RNA via two-color microarray analysis for a total of 20 arrays. In all cases, the experimental samples were labeled with Cy5 dye while the reference RNA was labeled with Cy3.

2009-11-30 | E-GEOD-8852 | biostudies-arrayexpress

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