The genomic analysis of endometrial mitochondrial DNA copy number variation on recurrent implantation failure

Aim of this study was to define the relationship between RIF (Recurrent Implantation Failure) and endometrial mtDNA copy number.

A total of 50 women of reproductive age including twenty-five patients clinically diagnosed with RIF and twenty-five fertile women as healthy controls were recruited into the study. Endometrial biopsy samples were obtained with a pipelle at the 20-24 days of the menstrual cycle of each participant. Total genomic DNA samples were isolated from endometrial tissues; MT-ND1 (mitochondrially encoded NADH dehydrogenase I) and MT-CO2 (mitochondrially encoded cytochrome C oxidase II) target genes were amplified by droplet digital PCR (ddPCR). Nuclear GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) gene was also used for study normalization. The study has been conducted between February 2019 and June 2019.

Droplet digital PCR results were analyzed in "QuantaSoft" software. The concentration amount (copies/μl) of each participant's mitochondrial gene was normalized according to the GAPDH gene concentrations as nuclear reference. mtDNA amounts were compared between RIF patients and healthy controls. Normalized data was statistically evaluated using Mann-Whitney U test and ROC curve analysis.

It was concluded that the mitochondrial target gene (MT-ND1 and MT-CO2) copy number amount of RIF patients was higher than the one obtained from the healthy group in endometrial tissues. It is thought that higher mtDNA copy number at the RIF group may be related to increased oxidative stress in the endometrium. This stress factors may influence receptivity negatively and cause implantation failure. The receptivity of the endometrium is associated with the number of mtDNA copies and difference can be used as a biomarker for receptivity analysis.