TY - JOUR

T1 - Loci cg06256735 and cg15815843 in the MFAP5 gene regulatory regions are hypomethylated in varicose veins apparently due to active demethylation

AU - Smetanina, Mariya A

AU - Korolenya, Valeria A

AU - Sipin, Fedor A

AU - Oscorbin, Igor P

AU - Sevostyanova, Kseniya S

AU - Gavrilov, Konstantin A

AU - Shevela, Andrey I

AU - Filipenko, Maxim L

N1 - This research was funded by the Russian Science Foundation [grant number 22-25-00832]. Illumina sequencing experiments were supported by the Program of Fundamental Scientific Research of the Russian Federation (PFSR RF) “Fundamental Basics of Health Preservation” [grant number 121031300045-2]. © 2024 The Author(s).

PY - 2024/5/29

Y1 - 2024/5/29

N2 - Varicose vein disease (VVD) is a common health problem worldwide. Microfibril-associated protein 5 (MFAP5) is one of the potential key players in its pathogenesis. Our previous microarray analysis revealed the cg06256735 and cg15815843 loci in the regulatory regions of the MFAP5 gene as hypomethylated in varicose veins which correlated with its up-regulation. The aim of this work was to validate preliminary microarray data, estimate the level of 5-hydroxymethylcytosine (5hmC) at these loci, and determine the methylation status of one of them in different layers of the venous wall. For this, methyl- and hydroxymethyl-sensitive restriction techniques were used followed by real-time PCR and droplet digital PCR, correspondingly, as well as bisulfite pyrosequencing of +/- oxidized DNA. Our microarray data on hypomethylation at the cg06256735 and cg15815843 loci in whole varicose vein segments were confirmed and it was also demonstrated that the level of 5hmC at these loci is increased in VVD. Specifically, among other layers of the venous wall, tunica (t.) intima is the main contributor to hypomethylation at the cg06256735 locus in varicose veins. Thus, it was shown that hypomethylation at the cg06256735 and cg15815843 loci takes place in VVD, with evidence to suggest that it happens through their active demethylation leading to up-regulation of the MFAP5 gene, and t. intima is most involved in this biochemical process.

AB - Varicose vein disease (VVD) is a common health problem worldwide. Microfibril-associated protein 5 (MFAP5) is one of the potential key players in its pathogenesis. Our previous microarray analysis revealed the cg06256735 and cg15815843 loci in the regulatory regions of the MFAP5 gene as hypomethylated in varicose veins which correlated with its up-regulation. The aim of this work was to validate preliminary microarray data, estimate the level of 5-hydroxymethylcytosine (5hmC) at these loci, and determine the methylation status of one of them in different layers of the venous wall. For this, methyl- and hydroxymethyl-sensitive restriction techniques were used followed by real-time PCR and droplet digital PCR, correspondingly, as well as bisulfite pyrosequencing of +/- oxidized DNA. Our microarray data on hypomethylation at the cg06256735 and cg15815843 loci in whole varicose vein segments were confirmed and it was also demonstrated that the level of 5hmC at these loci is increased in VVD. Specifically, among other layers of the venous wall, tunica (t.) intima is the main contributor to hypomethylation at the cg06256735 locus in varicose veins. Thus, it was shown that hypomethylation at the cg06256735 and cg15815843 loci takes place in VVD, with evidence to suggest that it happens through their active demethylation leading to up-regulation of the MFAP5 gene, and t. intima is most involved in this biochemical process.

KW - Adult

KW - Aged

KW - Female

KW - Humans

KW - Male

KW - Middle Aged

KW - 5-Methylcytosine/analogs & derivatives

KW - DNA Methylation

KW - Genetic Loci

KW - Regulatory Sequences, Nucleic Acid/genetics

KW - Varicose Veins/genetics

KW - Contractile Proteins/genetics

KW - Intercellular Signaling Peptides and Proteins/genetics

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85194975595&origin=inward&txGid=24f6ea387e10c24013d1bf6300457b73

U2 - 10.1042/BSR20231938

DO - 10.1042/BSR20231938

M3 - Article

C2 - 38743016

VL - 44

JO - Bioscience reports

JF - Bioscience reports

SN - 0144-8463

IS - 5

M1 - 44

ER -