Conjunctival Inflammatory Gene Expression Profiling in Dry Eye Disease: Correlations With HLA-DRA and HLA-DRB1

Abstract

Purpose: In several multicenter clinical trials, HLA-DR was found to be a potential biomarker of dry eye disease (DED)'s severity and prognosis. Given the fact that HLA-DR receptor is a heterodimer consisting in an alpha and a beta chain, we intended to investigate the correlation of inflammatory targets with the corresponding transcripts, HLA-DRA and HLA-DRB1, to characterize specific targets closely related to HLA-DR expressed in conjunctival cells from patients suffering from DED of various etiologies.Methods: A prospective study was conducted in 88 patients with different forms of DED. Ocular symptom scores, ocular-staining grades, tear breakup time (TBUT) and Schirmer test were evaluated. Superficial conjunctival cells were collected by impression cytology and total RNAs were extracted for analyses using the new NanoString® nCounter technology based on an inflammatory human code set containing 249 inflammatory genes.Results: Two hundred transcripts were reliably detected in conjunctival specimens at various levels ranging from 1 to 222,546 RNA copies. Overall, from the 88 samples, 21 target genes showed a highly significant correlation (R > 0.8) with HLA-DRA and HLA-DRB1, HLA-DRA and B1 presenting the highest correlation (R = 0.9). These selected targets belonged to eight family groups, namely interferon and interferon-stimulated genes, tumor necrosis factor superfamily and related factors, Toll-like receptors and related factors, complement system factors, chemokines/cytokines, the RIPK enzyme family, and transduction signals such as the STAT and MAPK families.Conclusions: We have identified a profile of 21 transcripts correlated with HLA-DR expression, suggesting closely regulated signaling pathways and possible direct or indirect interactions between them. The NanoString® nCounter technology in conjunctival imprints could constitute a reliable tool in the future for wider screening of inflammatory biomarkers in DED, usable in very small samples. Broader combinations of biomarkers associated with HLA-DR could be analyzed to develop new diagnostic approaches, identify tighter pathophysiological gene signatures and personalize DED therapies more efficiently.

Differentially Expressed Gene Pathways in the Conjunctiva of Sjögren Syndrome Keratoconjunctivitis Sicca

“…Our results showing increased XBP-1 and STAT1 levels agree with the literature and with our previous publication that showed increased XBP-1 and GRP78 in the SS KCS conjunctiva compared to healthy controls (11). Increased levels of STAT1 have been previously reported in humans and animal models, also in the conjunctiva (13,14,62,109). XBP-1 (X-box binding protein-1) participates in ER stress and UPR, decreasing salivary gland protein secretion (110).…”

“…Previous studies have identified a significant role for inflammation in the conjunctiva of SS KCS patients (6,13,14,34). To gain insight into the pathways involved, we analyzed which DEGs have been annotated or identified in immune pathways using the ROSALIND software and nCounter software, using the gene set analysis.…”

Effect of persimmon leaves (Diospyros kaki) on goblet cell density and inflammation in experimental dry eye model

“…4). Moreover, in human corneal epithelial cells, IRF-1 levels increased after a bacterial challenge, for example, Pseudomonas aeruginosa, which played an important role in MHC class II gene expression in macrophages [50]. However, interestingly, flavonoids selectively induce IRF-1 degradation in intestinal epithelial cells [51] in which there are many flavonoids in the EEDK.…”

State-of-the-art etiology and pathogenesis of dry eye

“…При помощи проточной цитометрии HLA-DR был обнаружен в эпителиальных клетках конъюнктивы у пациентов с ССГ. Авторами было высказано предположение о наличии прямой зависимости между количеством найденного HLA-DR и тяжестью течения ССГ, однако в ходе дальнейших исследований гипотеза не подтвердилась [50]. заключение В изученном материале предпринята попытка обобщения современных данных об этиологии и патогенезе ССГ.…”