Use of High Throughput Genomic Tools for the Study of Endothelial Cell Biology

Abstract

The endothelium is an active, dynamic and heterogeneous organ. It lines the vessels in every organ system and regulates diverse and important biological functions. Over the past several years researchers have gained enormous insights into endothelial cell function in physiological processes such as coagulation and vascular reactivity, and pathophysiological disease states such as inflammation and atherosclerosis. Despite our expanding knowledge of endothelial cell biology, the molecular mechanisms underlying these functions remain largely unknown. The newly developed high throughput genomic tools and accompanying analytical methods provide powerful approaches for identifying new endothelial cell genes and characterizing their role in health and disease. Here, we review some of the recent genomics and proteomic advances that are providing new methodologies for endothelial cell and vascular biology research.

Inflammatory Manifestations of Experimental Lymphatic Insufficiency

“…Transcriptional profiling has been utilized to identify genes activated in disease states and to refine targets for molecular therapy. Microarray technology has been applied to the elucidation of endothelial biology in health and disease [ 9 , 29 ], as well as to the investigation of gene expression patterns in cutaneous diseases [ 30 ] and in a wide array of non-neoplastic diseases that entail inflammatory or immune responses [ 31 ]. This approach is particularly attractive for the problem of acquired lymphedema, where the heterogeneous cellular composition of the tissues exposed to lymph stagnation presupposes a very complex, interdependent pattern of gene expression.…”

“…These observations were correlated with an assessment of the cutaneous histopathology in the lymphedema tissue. Furthermore, to investigate the molecular mechanisms of tissue response to lymphatic vascular insufficiency, we have undertaken a large-scale transcriptional profiling of the lymphedema tissue utilizing a comprehensive mouse cDNA microarray containing 42,300 features, representing over 25,000 unique genes and expressed sequence tags (ESTs) [ 9 ]. The patterns of gene expression in lymphedema were contrasted with those observed in normal and surgical sham controls.…”

Therapeutic Responses to Exogenous VEGF-C Administration in Experimental Lymphedema: Immunohistochemical and Molecular Characterization

“…Total RNA was isolated using a modified two-step purification protocol, as described previously. 21 RNA integrity was assessed using the Agilent 2100 Bioanalyzer System with RNA 6000 Pico LabChip Kit (Agilent, Palo Alto, California, USA). First-strand cDNA was synthesized from 15 g of total RNA derived from each pool and from whole embryonicday-17.5 embryo for reference RNA, in the presence of Cy3 and Cy5 dUTP, respectively, and hybridized to the Mouse Transcriptome Microarray.…”

A Novel Methodology to Probe Endothelial Differential Gene Expression Profile Reveals Novel Genes

“…Major obstacles include difficulty obtaining a single population of endothelial cell type, as well as the very small amounts of RNA extractable from endothelial cells, insufficient for standard gene expression studies (Tabibiazar and Quertermous 2003). Our study is significant for development of methodology combining and modifying a number of recently developed advanced techniques, so as to identify differential gene expression in a clinical context, for comparison of an individual patient's endothelial cell gene expression with control.…”