Evidence of Mitophagy in Lens Capsule Epithelial Cells of Patients with Pseudoexfoliation Syndrome.

PRCIS: Alterations in the PINK-mediated mitophagy pathway play an important role in PEX disease. PURPOSE: Pseudoexfoliation Syndrome (PEX) is a condition in which aberrant fibrillary protein builds up in various components of the eye and other extraocular tissues. In this study, we aim to investigate the functionality of intracellular auto-degradative machinery -especially mitophagy- and related genes and proteins in PEX. METHODS: Anterior lens capsules were obtained from cataracts patients with and without PEX to constitute the PEX group and age-matched controls during microincision cataracts surgery. PINK1-mediated mitophagy markers were evaluated on the transcriptional and translational level via RT-qPCR and immunohistochemistry analysis, respectively. RESULTS: The lens epithelial cells of PEX patients were characterized by significantly higher PINK1 gene expression compared to that of the controls (P<0.05). In terms of intensity of staining of expressed proteins, PINK1 (P<0.05), Parkin (P<0.01) and LC3B (P<0.01) were all statistically higher in PEX, compared to the controls. CONCLUSION: Altered auto-degradative response -specifically mitophagy- is a component of increased oxidative stress in PEX patients. The role of this mechanism in emerging complications warrants further research.

Investigation of Mitophagy Biomarkers in Corneal Epithelium of Keratoconus Patients.

PURPOSE: The pathological mechanisms of keratoconus (KC) have not been elucidated yet. Mitophagy is an important mechanism that eliminates damaged mitochondria under oxidative stress, and it could be one of the leading pathological causes of KC. This study aimed to find out the role of mitophagy in the keratoconic corneal epithelium. METHODS: The corneal epithelia were collected from the 103 progressive KC patients and the 46 control subjects. The real-time quantitative PCR was performed for PTEN-putative kinase-1 (PINK1), PARKIN, p62, and BNIP3 gene expressions in 31 KC and 9 control subjects. Western blot analyses were performed to investigate the protein expressions of PINK1, PARKIN, LC3B, ATG5, and BECLIN in the remaining 109 corneal epithelium samples from 72 patients and 37 control subjects. RESULTS: mRNA and protein expressions of PINK1 decreased significantly in the corneal epithelium of KC patients compared to the control subjects. No significant change was found in mRNA levels of PARKIN, p62, and BNIP3 in KC patients. The protein expression of PARKIN, LC3B, ATG5, and Beclin did not significantly differ between KC patients and control subjects. Gene expression levels of mitophagy biomarkers were not affected by the KC grade. CONCLUSIONS: PINK1/PARKIN-dependent mitophagy is affected in the keratoconic corneal epithelium. We found significant decreases in both mRNA and protein expressions of PINK1 in the keratoconic corneal epithelium. However, we did not observe any other significant change in mitophagy markers. Mitochondrial stress-related mitophagy pathways could be interrupted by the decreased levels of PINK1 in the keratoconic corneal epithelium, but solely PINK1 dysregulation is not likely to induce KC pathogenesis.