Cited by 159Retinal Diseases and TreatmentsEpigenetics and DNA MethylationMitochondrial Function and Pathology

Epigenetic Modification of Mitochondrial DNA in the Development of Diabetic Retinopathy

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Mishra, Manish, Kowluru, Renu A.·2015·Investigative Opthalmology & Visual Science

Abstract

PURPOSE: Retinal mitochondria are dysfunctional in diabetes, and mitochondrial DNA (mtDNA) is damaged and its transcription is compromised. Our aim was to investigate the role of mtDNA methylation in the development of diabetic retinopathy. METHODS: Effect of high glucose (20 mM) on mtDNA methylation was analyzed in retinal endothelial cells by methylation-specific PCR and by quantifying 5-methylcytosine (5mC). Dnmt1 binding at the D-loop and Cytb regions of mtDNA was analyzed by chromatin immunoprecipitation. The role of mtDNA methylation in transcription and cell death was confirmed by quantifying transcripts of mtDNA-encoded genes (Cytb, ND6, and CoxII) and apoptosis, using cells transfected with Dnmt1-small interfering RNA (siRNA), or incubated with a Dnmt inhibitor. The key parameters were validated in the retinal microvasculature from human donors with diabetic retinopathy. RESULTS: High glucose increased mtDNA methylation, and methylation was significantly higher at the D-loop than at the Cytb and CoxII regions. Mitochondrial accumulation of Dnmt1 and its binding at the D-loop were also significantly increased. Inhibition of Dnmt by its siRNA or pharmacologic inhibitor ameliorated glucose-induced increase in 5mC levels and cell apoptosis. Retinal microvasculature from human donors with diabetic retinopathy presented similar increase in D-loop methylation and decrease in mtDNA transcription. CONCLUSIONS: Hypermethylation of mtDNA in diabetes impairs its transcription, resulting in dysfunctional mitochondria and accelerated capillary cell apoptosis. Regulation of mtDNA methylation has potential to restore mitochondrial homeostasis and inhibit/retard the development of diabetic retinopathy.

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  • Factors including traffic-related elemental carbon, maternal smoking, endogenous metabolites, cell senescence, and ageing alter mtDNA methylation.

    “Furthermore, mtDNA methylation is disturbed by environmental and other factors, such as traffic-derived elemental carbon105, maternal smoking106, endogenous metabolites107, cell senescence108 and ageing109.”

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Authors
Mishra, M., & Kowluru, R. A.
Affiliations
Wayne State University (US)
Corresponding author
Kowluru, Renu A.
Venue
Investigative Opthalmology & Visual Science
Published
2015-08-04
Volume
56
Issue
9
Pages
5133
ISSN
  • 0146-0404 (ISSN-L)
  • 1552-5783
OpenAlex
W1531836650
Cited by
159
References
51
Citations by year
2026: 11 · 2025: 12 · 2024: 12 · 2023: 16 · 2022: 11 · 2021: 14 · 2020: 20 · 2019: 16 · 2018: 15 · 2017: 16 · 2016: 14 · 2015: 2
FWCI
15.04
Type
journal article
Language
en
Open access
Locations
  • Investigative Ophthalmology & Visual Science (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Research to Prevent Blindness
  • Wayne State University
  • National Institutes of Health
  • National Institute of Diabetes and Digestive and Kidney Diseases (P30DK020572)
Topics
  • Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
Keywords
diabetic retinopathy, mitochondrial DNA methylation, DNMT1, 5-methylcytosine, high glucose, retinal endothelial cells, mtDNA transcription, apoptosis, mitochondrial dysfunction, retinal microvasculature
MeSH terms
Aged, Animals, Cattle, Cells, Cultured, Diabetes Mellitus, Experimental, Diabetic Retinopathy, DNA Damage, DNA, Mitochondrial, Humans, Middle Aged, Mitochondria, Transcription Factors, Polymerase Chain Reaction, Apoptosis, Epigenesis, Genetic

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