MinION nanopore sequencing provides similar methylation estimates to Sanger bisulfite sequencing in the TRPA1 promoter region

Gombert, Sara; Jahn, Kirsten; Pathak, Hansi; Burkert, Alexandra; Schmidt, Gunnar; Wiehlmann, Lutz; Davenport, Colin; Braendl, Bjoern; Mueller, Franz-Josef; Leffler, Andreas; Deest, Maximilian; Frieling, Helge

doi:10.1101/2021.09.17.460763

Cited by 1 publication

(1 citation statement)

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“…Sanger sequencing has some upsides, such as still being the gold standard of comparison as well as the effective investigation of specific genetic loci. Concerning the method’ s accuracy for measuring single promoter regions, our research group showed that Sanger sequencing achieves comparable results compared with (ONT-S) when analyzing promoter methylation [55].…”

Section: Discussionmentioning

confidence: 99%

Methylation of the Oxytocin, Oxytocin Receptor, and Vasopressin Gene Promoters in Tobacco Use Disorder during Cessation

Proskynitopoulos,

Bleich,

Muschler

et al. 2024

Neuropsychobiology

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Introduction: Vasopressin (AVP) and oxytocin (OT) exert sex-specific effects on social pair bonding and stress reactions while also influencing craving in substance use disorders. In this regard, intranasal oxytocin (OT) and AVP antagonists present potential treatments for tobacco use disorder (TUD). Since transcription of both hormones is also regulated by gene methylation, we hypothesized sex-specific changes in methylation levels of the AVP, OT, and OT receptor (OXTR) gene during nicotine withdrawal. Methods: The study population consisted of 49 smokers (29 males, 20 females) and 51 healthy non-smokers (25 males, 26 females). Blood was drawn at day 1, day 7, and day 14 of smoking cessation. Craving was assessed with the questionnaire on smoking urges (QSU). Results: Throughout cessation, mean methylation of the OT promoter gene increased in males and decreased in females. OXTR receptor methylation decreased in females, while in males it was significantly lower at day 7. Regarding the AVP promoter, mean methylation increased in males while there were no changes in females. Using mixed linear modeling, CpG position, time point, sex, and the interaction of time point and sex as well as time point, sex, and QSU had a significant fixed effect on OT and AVP gene methylation. The interaction effect suggests that sex, time point, and QSU are interrelated, meaning that, depending on the sex, methylation could be different at different time points and vice versa. There was no significant effect of QSU on mean OXTR methylation. Discussion: We identified differences at specific CpGs between controls and smokers in OT and AVP and in overall methylation of the AVP gene. Furthermore, we found sex-specific changes in mean methylation levels of the mentioned genes throughout smoking cessation, underlining the relevance of sex in the OT and vasopressin system. This is the first study on epigenetic regulation of the OT promoter in TUD. Our results have implications for research on the utility of the AVP and OT system for treating substance craving. Future studies on both targets need to analyze their effect in the context of sex, social factors, and gene regulation.

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Section: Discussionmentioning

confidence: 99%