Human primary retinal cells as an in-vitro model for investigating defective signalling caused by OPTN mutants associated with glaucoma

摘要: Abstract Studies carried out on the pathogenesis of glaucoma using murine cell lines and animal models require to be validated in human cells. Therefore, we explored possibility primary retinal cells (hPRCs) culture as a model for molecular studies testing potential therapeutic drugs. For this purpose, central tissue, obtained from enucleated eyes patients with anterior staphyloma, was digested trypsin grown medium containing supplements (basic fibroblast growth factor fetal bovine serum). hPRCs at passage 1 2, show expression either GFAP, glial marker, or β-III tubulin, ganglion (RGC)-specific marker. But passages 3–5 nearly all express several RGC-specific markers (Brn3 proteins, Thy-1, RBPMS NeuN) but not GFAP. Expression these indicated that may have functional properties RGCs. As RGCs are sensitive glaucoma-associated mutants OPTN, analysed survival upon overexpression OPTN mutants. Glaucoma-associated mutants, E50K-OPTN M98K-OPTN, induced significantly higher death compared WT-OPTN, whereas an amyotrophic lateral sclerosis-associated mutant, E478G-OPTN, did not. TBK1 inhibitor Amlexanox protected M98K-OPTN death. suppressed by inhibitors CaMKKβ AMPK well 661W, mouse line expresses RGC precursor Our results suggest under appropriate condition RGC-like properties. These can used explore mechanisms relevant cytoprotective compounds.