Landscape of microRNA in the aqueous humour of proliferative diabetic retinopathy as assessed by next‐generation sequencing

Abstract

Background The microRNAs (miRNA) have been found to play an important role in the pathogenesis of diabetic retinopathy. We try to explore the miRNA and piwi‐interacting RNA (piRNA) profile in the aqueous humour of proliferative diabetic retinopathy (PDR) using next‐generation sequencing (NGS). Methods Aqueous humour samples were collected from nine PDR eyes and nine cataract control eyes, and NGS was performed. Quantitative polymerase chain reaction (qPCR) was used to validate the sequencing results. An oxygen‐induced retinopathy (OIR) model was used to validate the angiogenesis related miRNA. Results In total, 484 miRNAs were differently expressed between the PDR eyes and cataract control eyes, including 210 mature miRNAs and 274 novel miRNAs. Furthermore, eight miRNAs and 30 piRNAs were identified as the most differently expressed between the two groups (P > .85). This differential expression of miRNA was predicted to regulate Rho protein signal transduction, neurotransmitter uptake and histone lysine methylation. Relative expression patterns of miR‐184, ‐150‐5p and ‐93‐5p were confirmed by qPCR. A reduced expression of miR‐93‐5p was confirmed in the OIR model. Conclusions This study comprehensively demonstrated the miRNA and piRNA expression profile of the aqueous humour of PDR eyes, which may serve as a potential biomarker and involved in the pathogenesis of PDR.

MicroRNA Expression in the Aqueous Humor of Patients with Diabetic Macular Edema

“…The peculiarity of the present study starts from the population examined: as a matter of fact, AH miRNAs were evaluated in T2D patients affected by DME. Previous studies evaluating miRNA expression through intraocular sampling are few in number and all but one were conducted on patients affected by PDR, in the context of either type 1 or type 2 diabetes [ 16 , 17 , 18 , 19 , 20 ]. We enrolled only patients showing DME with mild or moderate NPDR in an attempt to minimize the ischemic component and consequently the molecular pathways driven by ischemia.…”

“…Mammadzada et al [ 19 ] showed a significant increase in the miRNA-19a, miR-27a, miR-20a and miR-93 expression in PDR patients undergoing PPV as compared to the controls. Recently by next generation sequencing, Chen et al [ 20 ] found seven miRNAs upregulated (miR-150-5p, miR-30c-5p, miR16-2-3p, miR-1827, miR-140-3p and miR-93-5p) and one downregulated (miR-99-5p) in AH of T2D patients with PDR as compared to controls.…”

“…The four previous studies evaluating miRNA expression in the vitreous of PDR patients relied on patients affected by macular hole as the control group [ 16 , 17 , 18 , 19 ]. Chen et al [ 20 ], in the evaluation of miRNAs expression of PDR patients, and Cho et al [ 22 ], in the evaluation of miRNA expression in the AH of DME patients, used cataract patients as the control group. Therefore, this is the first study to evaluate intraocular miRNA expression in diabetic patients without DR and/or DME.…”

“…Gomaa et al [ 16 ] sampled the vitreous, evaluating miR-200b expression in both T1D and T2D patients, while Usui-Ouchi et al [ 17 ], Hirota et al [ 18 ] and Mammadzada et al [ 19 ] evaluated the expression of multiple miRNAs through a microarray or PCR array vitreal profile. Recently, Chen et al [ 20 ] performed next generation sequencing to evaluate multiple miRNA expression in AH of T2D patients. In the above-mentioned vitreal studies and in the aqueous study, patients recruited were affected by proliferative diabetic retinopathy (PDR).…”

Identification of Diagnostic and Prognostic microRNAs for Recurrent Vitreous Hemorrhage in Patients with Proliferative Diabetic Retinopathy

“…The association of miRNA-20a and -93 in PDR have previously been proposed [41]. Chen et al analyzed the aqueous humor of PDR patients in comparison to nonangiogenic controls, finding 484 miRNAs differentially expressed with only eight miRNAs significantly different (miRNA-16-2, -30c, -93, -99b, -140, -150, -1827, novel_mir4, novel_mir340).…”