The Interplay Between Metabolites and MicroRNAs in Aqueous Humor to Coordinate Corneal Endothelium Integrity.
Ueno Morio, Yoshii Kengo, Yamashita Tomoko, Sonomura Kazuhiro, Asada Kazuko, Ito Eiko, Fujita Tomoko, Sotozono Chie, Kinoshita Shigeru, Hamuro Junji
AI Summary
This study found specific aqueous humor metabolites (e.g., HIB) and microRNAs (e.g., miR-34a-5p) interact, influencing corneal endothelial health. This interplay is crucial for maintaining corneal clarity in diseases like bullous keratopathy.
Abstract
Purpose
The purpose of the study was to clarify the interplay between metabolites and microRNAs (miRs) in the aqueous humor (AqH) of bullous keratopathy (BK) patients to retain human corneal endothelium (HCE) integrity.
Design
Prospective, comparative, observational study.
Participants
A total of 55 patients with BK and 31 patients with cataract (Cat) as control.
Methods
A biostatic analysis of miRs and metabolites in the AqH, hierarchical clustering, and a least absolute shrinkage and selection operator (Lasso) analysis were employed. The miR levels in AqH of BK (n = 18) and Cat (n = 8) patients were determined using 3D-Gene human miR chips. Hierarchical clusters of metabolites detected by liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry in AqH specimens from 2 disease groups, BK (total n = 55) and Cat (total n = 31), were analyzed twice to confirm the reproducibility. The analytical procedure applied for investigating the association between metabolites and miRs in AqH was the exploratory data analysis of biostatistics to avoid any kind of prejudice. This research procedure includes a heat-map, cluster analysis, feature extraction techniques by principal component analysis, and a regression analysis method by Lasso. The cellular and released miR levels were validated using reverse transcription polymerase chain reaction and mitochondria membrane potential was assessed to determine the functional features of the released miRs.
Main outcome measures
Identification of interacting metabolites and miRs in AqH attenuating HCE degeneration.
Results
The metabolites that decreased in the AqH of BK patients revealed that 3-hydroxyisobutyric acid (HIB), 2-aminobutyric acid (AB) and branched-chain amino acids, and serine were categorized into the same cluster by hierarchical clustering of metabolites. The positive association of HIB with miR-34a-5p was confirmed ( P = 0.018), and the Lasso analysis identified the interplay between miR-34a-5p and HIB, between miR-24-3p and AB, and between miR-34c-5p and serine ( P = 0.041, 0.027, and 0.009, respectively). 3-hydroxyisobutyric acid upregulated the cellular miR-34a expression, mitochondrial membrane potential, and release of miR-184 in dedifferentiated cultured HCE cells.
Conclusions
Metabolites and miRs in AqH may synchronize in ensuring the integrity of the HCE to maintain efficient dehydration from the stroma.
Financial disclosures: Proprietary or commercial disclosure may be found after the references.
Shields Classification
Key Concepts5
The metabolites 3-hydroxyisobutyric acid (HIB), 2-aminobutyric acid (AB), branched-chain amino acids, and serine were decreased in the aqueous humor (AqH) of bullous keratopathy (BK) patients and categorized into the same cluster by hierarchical clustering.
A positive association was confirmed between 3-hydroxyisobutyric acid (HIB) and miR-34a-5p (P = 0.018) in the aqueous humor of bullous keratopathy patients.
Lasso analysis identified an interplay between miR-34a-5p and 3-hydroxyisobutyric acid (HIB) (P = 0.041), between miR-24-3p and 2-aminobutyric acid (AB) (P = 0.027), and between miR-34c-5p and serine (P = 0.009) in the aqueous humor of bullous keratopathy patients.
3-hydroxyisobutyric acid (HIB) upregulated the cellular miR-34a expression, mitochondrial membrane potential, and release of miR-184 in dedifferentiated cultured human corneal endothelium (HCE) cells.
Metabolites and microRNAs (miRs) in aqueous humor (AqH) may synchronize in ensuring the integrity of the human corneal endothelium (HCE) to maintain efficient dehydration from the stroma.
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