Genetic amplification of the transcriptional response to hypoxia as a novel means of identifying regulators of angiogenesis.
The cellular response to hypoxia involves the promotion of angiogenesis, leading to increased blood flow and oxygenation. The macrophage has been identified as an orchestrator of this response in several pathologies, through the release of angiogenic factors in response to hypoxia. We have produced the first comprehensive transcriptome analysis of hypoxic primary human macrophages with respect to the regulation of angiogenesis. There is a marked induction of genes encoding factors known to stimulate angiogenesis, rather than factors that inhibit this process. We show that overexpression of the transcription factor EPAS1 using a recombinant adenoviral vector amplifies the induction of genes encoding angiogenic proteins in response to hypoxia. This defines a new strategy for enhancing transcriptome and proteome analyses by overexpressing disease-implicated genes using viral gene transfer methodologies.
White
Jonathan Richard
JR
Biological Systems Group, Oxford BioMedica Ltd., OX4 4GA, Oxford, United Kingdom. j.white@oxfordbiomedica.co.uk
Harris
Robert A
RA
Lee
Sheena R
SR
Craigon
Marie H
MH
Binley
Katie
K
Price
Toby
T
Beard
Georgina L
GL
Mundy
Christopher R
CR
Naylor
S
S
D000256:Adenoviridae / Q000235:genetics; D051792:Basic Helix-Loop-Helix Proteins; D015687:Cell Hypoxia / Q000502:physiology; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D005809:Genes, Regulator / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D018919:Neovascularization, Physiologic / Q000235:genetics*; D012313:RNA / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics*; D000099162:Endothelial PAS Domain-Containing Protein 1
D051792:Basic Helix-Loop-Helix Proteins; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012313:RNA; D015534:Trans-Activators; D014157:Transcription Factors; D000099162:Endothelial PAS Domain-Containing Protein 1; C497442:HIF1A protein, human