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DNA Methylation—a Potential Source of Mitochondria DNA Base Mismatch in the Development of Diabetic Retinopathy

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Abstract

In the development of diabetic retinopathy, retinal mitochondria are dysfunctional, and mitochondrial DNA (mtDNA) is damaged with increased base mismatches and hypermethylated cytosines. DNA methylation is also a potential source of mutation, and in diabetes, the noncoding region, the displacement loop (D-loop), experiences more methylation and base mismatches than other regions of the mtDNA. Our aim was to investigate a possible crosstalk between mtDNA methylation and base mismatches in the development of diabetic retinopathy. The effect of inhibition of Dnmts (by 5-aza-2′-deoxycytidine or Dnmt1-siRNA) on glucose-induced mtDNA base mismatches was investigated in human retinal endothelial cells by surveyor endonuclease digestion and validated by Sanger sequencing. The role of deamination factors on increased base mismatches was determined in the cells genetically modulated for mitochondrial superoxide dismutase (Sod2) or cytidine-deaminase (APOBEC3A). The results were confirmed in an in vivo model using retinal microvasculature from diabetic mice overexpressing Sod2. Inhibition of DNA methylation, or regulation of cytosine deamination, significantly inhibited an increase in base mismatches at the D-loop and prevented mitochondrial dysfunction. Overexpression of Sod2 in mice also prevented diabetes-induced D-loop hypermethylation and increase in base mismatches. The crosstalk between DNA methylation and base mismatches continued even after termination of hyperglycemia, suggesting its role in the metabolic memory phenomenon associated with the progression of diabetic retinopathy. Inhibition of DNA methylation limits the availability of methylated cytosine for deamination, suggesting a crosstalk between DNA methylation and base mismatches. Thus, regulation of DNA methylation, or its deamination, should impede the development of diabetic retinopathy by preventing formation of base mismatches and mitochondrial dysfunction.

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Funding

The authors acknowledge the grant support from the National Institutes of Health (R01-EY014370, R01-EY017313, and R01-EY022230) and the Thomas Foundation to RAK and from Research to Prevent Blindness to the Ophthalmology Department.

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  1. Kresge Eye Institute, Wayne State University, Detroit, MI, 48201, USA

    Manish Mishra & Renu A. Kowluru

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Correspondence to Renu A. Kowluru.

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Mishra, M., Kowluru, R.A. DNA Methylation—a Potential Source of Mitochondria DNA Base Mismatch in the Development of Diabetic Retinopathy. Mol Neurobiol 56, 88–101 (2019). https://doi.org/10.1007/s12035-018-1086-9

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