Deduction of Novel Genes Potentially Involved in the Effects of Very Low Dose Atropine (0.003%) Treatment on Corneal Epithelial Cells Using Next-Generation Sequencing and Bioinformatics Approaches

Abstract

Background and Objectives: Atropine is a nonselective muscarinic antagonist which has been used to prevent worsening of myopia in children. Different concentrations of atropine were used for myopia, ranging from 0.01% to 1.0%. However, there are still potential toxicity of different doses of atropine to the cornea. Here, we present a study of investigating novel genes potentially involved in the effects of very low dose atropine treatment (0.003%) on corneal epithelial cells using next-generation sequencing (NGS) and bioinformatics approaches. Materials and Methods: Human corneal epithelial cells were treated with 0.003% atropine, cultured until confluence, and RNA extracted for differential expression profiling of mRNA and microRNA (miRNA) between control and atropine-treated corneal epithelial cells. The functional enrichment analysis for differentially expressed genes was performed using two bioinformatics databases, including Database for Annotation, Visualization and Integrated Discovery (DAVID) and Ingenuity® Pathway Analysis (IPA). In addition, potential miRNA-mRNA interactions involved in atropine-treated corneal epithelial cells were predicted and validated using different miRNA target prediction databases. Results: Our results showed 0.003% atropine might suppress the apoptosis of corneal epithelial cells, potentially through Ras and protein kinase A signaling pathways. We also validated the possible miRNA regulations by using TargetScan and miRDB databases. Hsa-miR-651-3p-EPHA7, hsa-miR-3148-TMEM108 and hsa-miR-874-5p-TBX6 were validated as possible miRNA regulations involved in corneal epithelial cells treated with 0.003% atropine. Conclusions: These findings may contribute novel insights into therapeutic strategies for treating cornea with 0.003% atropine.

IMI 2021 Yearly Digest

“… 118 Another genetic study similarly derived a PRS for refractive error that attained an AUC of 0.75 for predicting high myopia and found that children with a PRS in the top 10% were at 6-fold higher risk of high myopia than those in the remaining 90%. 119 This latter study was based on a much smaller sample than Hysi et al., 111 suggesting that there is hope for even greater accuracy in genetic prediction of myopia in the future. The genetic loci associated with myopia have relatively small effect sizes, so identifying more loci will not necessarily improve prediction of common myopia.…”

“…Two reports on the novel topic of miRNA expression in low-dose atropine treatment (0.003%) against myopia development were published. 111 , 112 Both studies focused on gaining more insight into the molecular mechanism of atropine on myopia treatment and defending the safety of low-dose atropine treatment, using either human scleral fibroblasts 112 or human corneal epithelial cells. 111 Hsiao et al.…”

“… 111 , 112 Both studies focused on gaining more insight into the molecular mechanism of atropine on myopia treatment and defending the safety of low-dose atropine treatment, using either human scleral fibroblasts 112 or human corneal epithelial cells. 111 Hsiao et al. 112 identified slight changes in scleral gene expression after 0.003% atropine treatment, supporting the safety of low-dose atropine treatment.…”

“…Chang et al. 111 analyzed the messenger RNA (mRNA) and miRNA expression profiles between atropine-treated and control corneal epithelial cells. They found that low-dose (0.003%) atropine was associated with dysregulation of certain genes.…”

Corneal Penetration of Low-Dose Atropine Eye Drops

“…With increasing levels, an increasing probability of off-target effects must also be assumed [ 28 ]. It has been speculated that, in addition to the retina, scleral fibroblasts may also be involved [ 29 , 30 ], or, more unlikely, the corneal endothelium [ 31 ]. It was identified early that pharmacodynamic effects also depend on iris pigmentation: mydriasis lasted significantly longer in pigmented animals (half-life 96 h vs. 43 h in pigmented vs. albino animals) [ 32 ].…”