Microarray analysis of gene expression in human donor corneas

Objectives: To use microarray analysis to identify genes expressed in human donor corneas and to create a preliminary, comprehensive database of human corneal gene expression. Methods: A complementary DNA (cDNA) library was constructed from transplant -quality, human donor corneas. Biotin-labeled RNA was transcribed from the cDNA library and hybridized in duplicate to microarrays containing approxim ately 5600 human genes. Results were analyzed using a gene database of the National Institutes of Health, Bethesda, Md. Reverse transcriptase polymera se chain reaction analysis was performed to confirm corneal expression of g enes identified by microarray analysis. Results: Duplicate microarrays identified the expression of 1200 genes in h uman donor corneas. Chromosomal loci had been assigned to 1025 (85%) of the se genes. A preliminary database of human corneal gene expression was compi led. A Web site containing these genes was created. Six collagen genes were identified that had not previously been localized within the cornea. Five apoptosis-related genes were identified, 4 of which had not previously been localized within the cornea. Three genes previously shown to cause corneal diseases were identified. Reverse transcriptase polymerase chain reaction analysis of genes identified by microarray analysis confirmed the corneal e xpression of 2 apoptosis-related genes and 1 collagen gene. Conclusions: Microarray analysis of healthy human donor corneas has produce d a preliminary, comprehensive database of corneal gene expression. Large-s cale analysis of gene expression has the potential to generate large amount s of data, which should be made readily accessible to the scientific commun ity. The Internet offers many potential advantages as a medium for the main tenance of these large data sets. Clinical Relevance: Identification of structural, apoptosis-related, and di sease-causing genes within the cornea by microarrays may increase the under standing of normal and abnormal corneal function with likely relevance to c orneal diseases and transplants.