Genome-wide expression profile of human trabecular meshwork cultured cells, nonglaucomatous and primary open angle glaucoma tissue.
OBJECTIVE
To contrast genome-wide gene expression profiles of cultured human trabecular meshwork (HTM) cells to that of control and primary open angle glaucoma (POAG) HTM tissues.


METHODS
Cultured HTM cells, HTM tissue dissected from control donors, and HTM tissue from POAG donors receiving medication for glaucoma were fixed in RNA latertrade mark. Total RNA extracted from these samples was linearly amplified with the Ovation Biotin RNA Amplification and Labeling System and individually hybridized to Affymetrix Human Genome U133 Plus 2.0 high density microarrays. Data analysis was performed using GeneSpring Software 7.0. Selected genes showing significant differential expression were validated by quantitative real-time PCR in nonamplified RNA.


RESULTS
Cultured HTM cells retained the expression of some genes characteristic of HTM tissue, including chitinase 3-like 1 and matrix Gla protein, but demonstrated downregulation of physiologically important genes such as myocilin. POAG HTM tissue showed relatively small changes compared to that of control donors. These changes included the statistically significant upregulation of several genes associated with inflammation and acute-phase response, including selectin-E (ELAM-I), as well as the downregulation of the antioxidants paraoxonase 3 and ceruloplasmin.


CONCLUSIONS
Downregulation in cultured HTM cells of genes potentially relevant for outflow pathway function highlights the importance of developing new conditions for the culture of TM cells capable of preserving the characteristics of TM cells in vivo. Comparative analysis between control and POAG tissues suggests that the upregulation of inflammation-associated genes might be involved in the progression of glaucoma.
Liton
Paloma B
PB
Department of Ophthalmology, Duke University, Durham, NC 27710, USA.
Luna
Coralia
C
Challa
Pratap
P
Epstein
David L
DL
Gonzalez
Pedro
P
D016022:Case-Control Studies; D002478:Cells, Cultured; D002874:Chromosome Mapping; D015536:Down-Regulation; D020869:Gene Expression Profiling; D005902:Glaucoma, Open-Angle / Q000235:genetics*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D014129:Trabecular Meshwork / Q000378:metabolism* / Q000473:pathology; D015854:Up-Regulation