Novel Genes Correlating to in vivo Retinal Vessel Loss and Regrowth Identified Through a New High Sensitivity Analysis of Gene Microarrays

Abstract

Abstract: : Purpose: An early manifestation of many diseases is loss or proliferation of blood vessels. Serum vascular markers have recently been found to predict later onset of type 2 diabetes. A larger array of vascular markers is likely to allow more specific detection of many diseases. Our aim was to determine novel retinal vascular markers for possible early detection of eye disease Methods: P7 mice exposed to hyperoxia have extensive loss of retinal vessels without initial loss of neural retina. Gene expression changes at P7–P12 between age–matched normal retina and retina with hyperoxia–induced vascular loss (reflecting primarily vessel loss) were examined with Affymetrix microarrays. Data was analyzed using a new Bayesian method to determine sources of noise in the microarray data to predict which genes would validate with real time RT–PCR. Results: Twelve genes were found which validated with real time RT–PCR and had decreased expression with vascular loss, increased expression with vaso–proliferation and were expressed in endothelial or smooth muscle cell culture. These were transmembrane 4 superfamily member 1 (Tm4sf1); basigin (Bsg); claudin–5 (Cldn5); EGF, latrophilin seven transmembrane domain containing 1 (Eltd1); occludin (Ocln); biglycan (Bgn); matrix gamma–carboxyglutamate protein (Mglap); macrophage migration inhibitory factor (Mif); neural precursor cell expressed, developmentally down–regulated gene 4–like (Nedd4l); regulator of G–protein signaling 5 (Rgs5); PDZ and LIM domain 3 (Pdlim3). Conclusions: Expression changes in these genes are predictive of vessel loss and re–growth and are potential surrogate markers of early changes in eye diseases such as diabetic retinopathy or cancer.

Keywords: diabetic retinopathy • gene microarray • retinal neovascularization