Differential global and extra-cellular matrix focused gene expression patterns between normal and glaucomatous human lamina cribrosa cells

[摘要] Purpose: Marked extracellular matrix(ECM) remodeling occurs in the human optic nerve head in primary openangle glaucoma (POAG). The glial fibrillary acid protein (GFAP)negative lamina cribrosa cell may play an important role in thisremodeling process. We report the first study of global and ECM-focusedgene transcription differentials between GFAP-negative lamina cribrosa(LC) cells from normal and POAG human donors. Methods: GFAP-negative LC cell lineswere generated from the optic nerve tissue of four normal (n=4) andfour POAG (n=4) human donors. Using Affymetrix U133A arrays thetranscriptional profile between the normal and diseased groups werecompared. Bioinformatic analysis was performed using robust multichipaverage (RMA Express) and EASE/David. Real time TaqMan PCR andimmunohistochemistry analyses were performed to validate the microarraydata. Results: 183 genes were upregulated bygreater than 1.5 fold and 220 were down regulated by greater than 1.5fold in the POAG LC cells versus normal controls. Upregulated genes inPOAG LC cells included, transforming growth factor beta 1 (TGFβ1),secreted acid protein cysteine rich (SPARC), periostin (POSTN),thrombospondin-1 (THBS1), cartilage linking protein-1 (CRTL-1),and collagen type I (COL1A1), collagen type V (COL5A1),and collagen type XI (COL11A1). Downregulated ECM genes in POAGincluded fibulin 1 (FBLN1), decorin (DCN), and collagentype XVIII (COL18A1). All TaqMan PCR validation assays weresignificant (*p<0.05) and consistent with the array data.Immunohistochemistry of one target (periostin) confirmed itsdifferential expression at the protein level in POAG optic nerve headtissue compared with non-glaucomatous controls. Functional annotationand over-representation analysis identified ECM genes as astatistically over-represented class of genes in POAG LC cells comparedwith normal LC cells. Conclusions: This study reports for thefirst time that POAG LC cells in-vitro demonstrate upregulated ECM andpro-fibrotic gene expression compared with normal LC cells. This may bea pathological characteristic of this cell type in POAG in-vivo. Webelieve that the LC cell may be a pivotal regulator of optic nerve headECM remodeling in POAG and an attractive target for moleculartherapeutic strategies in the future.

[学科分类] 生物化学/生物物理