Transforming growth factor-β-regulated gene transcription and protein expression in human GFAP-negative lamina cribrosa cells

• Published in: Glia 2005-12, Vol.52 (4), p.309-324
• Main Authors: Kirwan, Ruaidhrí P., Leonard, Martin O., Murphy, Madeline, Clark, Abbot F., O'Brien, Colm J.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Cells, Cultured; extracellular matrix; Extracellular Matrix - metabolism; Fibrosis - genetics; Fibrosis - metabolism; Fibrosis - prevention & control; Fundamental and applied biological sciences. Psychology; Gene Expression Profiling; Gene Expression Regulation - drug effects; Gene Expression Regulation - genetics; glaucoma; Glaucoma, Open-Angle - genetics; Glaucoma, Open-Angle - metabolism; Glaucoma, Open-Angle - physiopathology; Glial Fibrillary Acidic Protein - metabolism; Growth Substances - genetics; Growth Substances - metabolism; Humans; Isolated neuron and nerve. Neuroglia; lamina cribrosa; microarray; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Neuroglia - drug effects; Neuroglia - metabolism; Neuroglia - ultrastructure; Oligonucleotide Array Sequence Analysis; Optic Disk - drug effects; Optic Disk - metabolism; Optic Disk - ultrastructure; Polymerase Chain Reaction; Regulatory Elements, Transcriptional - genetics; RNA, Messenger - analysis; RNA, Messenger - genetics; TGF-β; Transcriptional Activation - drug effects; Transcriptional Activation - genetics; Transforming Growth Factor beta - metabolism; Transforming Growth Factor beta - pharmacology; Up-Regulation - drug effects; Up-Regulation - genetics; Vertebrates: nervous system and sense organs
• ISSN: 0894-1491; 1098-1136
• DOI: 10.1002/glia.20247

Primary open‐angle glaucoma (POAG) is a progressive optic neuropathy, which is a major cause of worldwide visual impairment and blindness. Pathological hallmarks of the glaucomatous optic nerve head (ONH) include retinal ganglion cell axon loss and extracellular matrix (ECM) remodeling of the lamina cribrosa layer. Transforming growth factor‐β (TGF‐β) is an important pro‐fibrotic modulator of ECM metabolism, whose levels are elevated in human POAG lamina cribrosa tissue compared with non‐glaucomatous controls. We hypothesize that in POAG, lamina cribrosa (LC) glial cells respond to elevated TGF‐β, producing a remodeled ONH ECM. Using Affymetrix microarrays, we report the first study examining the effect of TGF‐β1 on global gene expression profiles in glial fibrillary acidic acid (GFAP)‐negative LC glial cells in vitro. Prominent among the differentially expressed genes were those with established fibrogenic potential, including CTGF, collagen I, elastin, thrombospondin, decorin, biglycan, and fibromodulin. Independent TaqMan and Sybr Green quantitative PCR analysis significantly validated genes involved in regulation of cell proliferation (platelet‐derived growth factor [PDGF‐α]), angiogenesis (vascular endothelial growth factor [VEGF]), ECM accumulation and degradation (CTGF, IL‐11, and ADAMT‐S5), and growth factor binding (ESM‐1). Bioinformatic analysis of the ESM‐1 promoter identified putative Smad and Runx transcription factor binding sites, and luciferase assays confirmed that TGF‐β1 drives transcription of the ESM‐1 gene. TGF‐β1 induces expression and release of ECM components in LC cells, which may be important in regulating matrix remodeling in the lamina cribrosa. In disease states such as POAG, the LC cell may represent an important pro‐fibrotic cell type and an attractive target for novel therapeutic strategies. © 2005 Wiley‐Liss, Inc.