摘要：

Purpose : Topical glaucoma drugs are associated with a high prevalence of Ocular Surface Disease (OSD), significantly affecting patient adherence. Clinical manifestations of OSD resemble those observed in Dry Eye Syndrome (DES). However, the mechanisms mediating the onset of symptoms are unknown. We aim to identify conjunctival transcriptomic changes in both glaucoma and DES patients, comparing them to healthy controls. Methods : Impression cytology tests (EYEPRIM) were performed on the temporal conjunctiva of 140 eyes. 56 samples were selected for RNA isolation and sequencing (33 treated with glaucoma drugs, 9 with DES and 14 controls). RNA Isolation (Maxwell 16 RNA extraction kit, Promega), quantification (NanoDrop spectrophotometer, Thermo Fisher Scientific) and quality assessment (Bioanalyzer, Agilent Technologies) were done prior to RNA sequencing (NextSeq 500, Illumina). Ingenuity Pathway Analysis (IPA) was used to complete the biological information. Results : Glaucoma patients showed dysregulation in 200 genes, while DES patients exhibited dysregulation in 233 genes, when compared to controls. According to the IPA database, the up-regulation of immune related genes in treated glaucoma patients represented 79% of the altered pathways. Dysregulated genes included cellular damage sensors: TLR4 (p<0.001), LY86 (p<0.001), P2RX7 (p=0.002); dendritic cells activation: TREM2 (p=0.003), CD86 (p<0.001) and Tec and Src Kinase pathways: BTK (p=0.002). In DES subjects, 39% of the modified pathways were linked to the immune system and inflammation: RGS1(p=0,001), RNASE6 (p=0,002), or FCER1G (p=0,001); 16% represented increased expression of genes related to mitochondrial metabolism and oxidative stress (MT-CYB (p=0,03), MT-CO2 (p<0,001) and 15% represented decreased expression of genes involved in maintaining conjunctival integrity and cell differentiation: YAP1 (p=0.009), TEAD1 (p=0.005), SMAD4 (p=0.006). TGFB pathway: ITGFB (p=0,003). Conclusions : According to the results, an exacerbation of the immune system is observed in patients with glaucoma treatment as the main pathway to ocular surface damage. However, in the case of patients with DES, the damage is the result of multiple pathways. Knowledge of the mechanisms involved in ocular surface damage is essential for a personalized approach to this issue.

展开