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Genome-wide analysis of the endothelial transcriptome under short-term chronic hypoxia

¹ Division of Pulmonary, Allergy and Critical Care Medicine
² Lung Translational Genomics Center, Department of Medicine, University of Pittsburgh, Pennsylvania 15213
³ Institute for Human and Machine Cognition, University of West Florida, Pensacola, Florida 32514

We have utilized serial analysis of gene expression (SAGE) to analyze the temporal response of human aortic endothelial cells (HAECs) to short-term chronic hypoxia at the level of transcription. Primary cultures of HAECs were exposed to 1% O₂ hypoxia for 8 and 24 h and compared with identical same passage cells cultured under standard (5% CO₂-95% air) conditions. A total of 121,446 tags representing 37,096 unique tags were sequenced and genes whose expression levels were modulated by hypoxia identified by novel statistical analyses. Hierarchical clustering of genes displaying statistically significant hypoxia-responsive alterations in expression revealed temporal modulation of a number of major functional gene families including those encoding heat shock factors, glycolytic enzymes, extracellular matrix factors, cytoskeletal factors, apoptotic factors, cell cycle regulators and angiogenic factors. Within these families we documented the coordinated modulation of both previously known hypoxia-responsive genes, numerous genes whose expressions have not been previously shown to be altered by hypoxia, tags matching uncharacterized UniGene entries and entirely novel tags with no UniGene match. These preliminary data, which indicate a reduction in cell cycle progression, elevated metabolic stress and increased cytoskeletal remodeling under acute hypoxic stress, provide a foundation for further analyses of the molecular mechanisms underlying the endothelial response to short-term chronic hypoxia.

endothelium; hypoxia; serial analysis of gene expression

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