RUDN Journal of MedicineRUDN Journal of MedicineВестник Российского университета дружбы народов. Серия: Медицина2313-02452313-0261Peoples’ Friendship University of Russia named after Patrice Lumumba (RUDN University)3717610.22363/2313-0245-2023-27-4-515-531IHQKOHMEDICAL GENETICSМЕДИЦИНСКАЯ ГЕНЕТИКАResearch ArticleAbnormal methylation of PRDM16 and PTPRN2 genes in chorionic villi in miscarriageНарушения метилирования генов PRDM16 и PTPRN2в ворсинах хориона при невынашивании беременностиhttps://orcid.org/0000-0002-5301-070X8087-5222VasilyevStanislav A.ВасильевС. А.stanislav.vasilyev@medgenetics.ruhttps://orcid.org/0000-0001-5797-00143582-1273VasilyevaOksana Yu.ВасильеваО. Ю.stanislav.vasilyev@medgenetics.ruOppong-PeprahBismarkОппонг-ПепрахБ.stanislav.vasilyev@medgenetics.ruhttps://orcid.org/0000-0002-5315-49143631-0953DemenevaVictoria V.ДеменеваВ. В.stanislav.vasilyev@medgenetics.ruhttps://orcid.org/0000-0001-9474-93353235-1754ZuevAndrey S.ЗуевА. С.stanislav.vasilyev@medgenetics.ruhttps://orcid.org/0000-0003-3875-39328788-4112SazhenovaElena A.СаженоваЕ. А.stanislav.vasilyev@medgenetics.ruhttps://orcid.org/0000-0002-4230-68558941-1605NikitinaTatiana V.НикитинаТ. В.stanislav.vasilyev@medgenetics.ruhttps://orcid.org/0000-0001-6427-32767837-4073TolmachevaEkaterina N.ТолмачеваЕ. Н.stanislav.vasilyev@medgenetics.ruTomsk National Research Medical CenterТомский национальный исследовательский медицинский центрNational Research Tomsk State UniversityНациональный исследовательский Томский государственный университет15122023274PHYSIOLOGY. EXPERIMENTAL PHYSIOLOGYФИЗИОЛОГИЯ. ЭКСПЕРИМЕНТАЛЬНАЯ ФИЗИОЛОГИЯ51553122122023Copyright ©; 2023, Vasilyev S.A., Vasilyeva O.Y., Oppong-Peprah B., Demeneva V.V., Zuev A.S., Sazhenova E.A., Nikitina T.V., Tolmacheva E.N.Copyright ©; 2023, Васильев С.А., Васильева О.Ю., Оппонг-Пепрах Б., Деменева В.В., Зуев А.С., Саженова Е.А., Никитина Т.В., Толмачева Е.Н.2023Vasilyev S.A., Vasilyeva O.Y., Oppong-Peprah B., Demeneva V.V., Zuev A.S., Sazhenova E.A., Nikitina T.V., Tolmacheva E.N.Васильев С.А., Васильева О.Ю., Оппонг-Пепрах Б., Деменева В.В., Зуев А.С., Саженова Е.А., Никитина Т.В., Толмачева Е.Н.https://creativecommons.org/licenses/by-nc/4.0https://journals.rudn.ru/medicine/article/view/37176

Relevance. Abnormal epigenetic regulation of genes responsible for the development of the embryo and placenta is associated with many pregnancy pathologies. Aim. The aim of this work was to analyze the prevalence of abnormal methylation of the PRDM16 and PTPRN2 genes in chorionic villi of spontaneous abortions with normal karyotype and with the most frequent aneuploidies (trisomy 16 and monosomy X). Materials and Methods. The methylation profile was evaluated using targeted bisulfite massive parallel sequencing in chorionic villi of induced abortions (n = 10), spontaneous abortions with normal karyotype (n = 39), trisomy 16 (n = 17) and monosomy X (n = 20) and peripheral blood lymphocytes of healthy volunteers (n = 6). Results and Discussion. In analyzed genes, differential methylation of individual CpG sites was found in chorionic villi of spontaneous abortions. Despite the absence of significant differences between the groups in the average level of methylation in analyzed gene regions, abnormal methylation of the PRDM16 and PTPRN2 genes were detected for 33 % and 5 % of spontaneous abortions, respectively, indicating a high incidence of epigenetic abnormalities in these genes in the chorionic villi of spontaneous abortions. The level of methylation of the PRDM16 gene significantly correlated with the level of methylation of the retrotransposon LINE-1, which indicates the generalized nature of methylation disorders in spontaneous abortions. Finally, the level of methylation of the PTPRN2 gene depended on the age of mothers of spontaneous abortions with monosomy X, which raises the question of the influence of maternal factors on the methylation profile in this group of spontaneous abortions. Conclusion. The results indicate that epigenetic disorders of the PRDM16 gene may be associated with spontaneous termination of pregnancy in the first trimester.

Актуальность. Нарушения эпигенетической регуляции генов, ответственных за развитие эмбриона и плаценты, ассоциированы со многими патологиями беременности. Цель. Целью настоящей работы стал анализ распространенности нарушений метилирования генов PRDM16 и PTPRN2 в ворсинах хориона спонтанных абортусов с нормальным кариотипом и с наиболее частыми анеуплоидиями (трисомия 16 и моносомия X). Материалы и методы. Оценка профиля метилирования была проведена с помощью таргетного бисульфитного массового параллельного секвенирования в ворсинах хориона медицинских абортусов (n = 10), спонтанных абортусов с нормальным кариотипом (n = 39), трисомией 16 (n = 17) и моносомией X (n = 20) и лимфоцитов периферической крови здоровых добровольцев (n = 6). Результаты и обсуждение. Было обнаружено дифференциальное метилирование отдельных CpG-сайтов в изученных генах в ворсинах хориона спонтанных абортусов. Несмотря на отсутствие значимых отличий между группами по среднему уровню метилирования в изученных регионах генов, отклонения уровня метилирования генов PRDM16 и PTPRN2 были выявлены для 33 % и 5 % спонтанных абортусов, соответственно, что указывает на высокую частоту распространения эпигенетических аномалий по этим генам в ворсинах хориона спонтанных абортусов. Уровень метилирования гена PRDM16 значимо коррелировал с уровнем метилирования ретротранспозона LINE-1, что указывает на генерализованный характер нарушений метилирования у спонтанных абортусов. Наконец, уровень метилирования гена PTPRN2 зависел от возраста матерей спонтанных абортусов с моносомией X, что поднимает вопрос о влиянии материнских факторов на профиль метилирования в этой группе спонтанных абортусов. Выводы . Полученные результаты указывают, что эпигенетические нарушения гена PRDM16 могут быть связаны со спонтанным прерыванием беременности в первом триместре.

PRDM16PTPRN2DNA methylationchorionic villimiscarriageaneuploidybisulfite sequencingspontaneous abortionsPRDM16PTPRN2метилирование ДНКворсины хорионаневынашивание беременностианеуплоидиябисульфитное секвенированиеспонтанные абортусыThe study was supported by the grant of the Russian Science Foundation No. 23–15–00341.Исследование выполнено при поддержке гранта Российского научного фонда № 23–15–00341.Edmonds DK, Lindsay KS, Miller JF, Williamson E, Wood PJ. Early embryonic mortality in women. Fertil Steril. 1982;38(4): 447-53.Ford HB, Schust DJ. Recurrent pregnancy loss: etiology, diagnosis, and therapy. Rev Obstet Gynecol. 2009;2(2):76-83.Eiben B, Bartels I, Bahr-Porsch S, Borgmann S, Gatz G, Gellert G, Goebel R, Hammans W, Hentemann M, Osmers R. Cytogenetic analysis of 750 spontaneous abortions with the direct-preparation method of chorionic villi and its implications for studying genetic causes of pregnancy wastage. Am J Hum Genet. 1990;47(4):656-63.Menasha J, Levy B, Hirschhorn K, Kardon NB. Incidence and spectrum of chromosome abnormalities in spontaneous abortions: new insights from a 12-year study. Genet Med. 2005;7(4): 251-63. doi: 00125817-200504000-00005Baranov VS, Kuznetsova TV. Cytogenetics of human embryonic development. 2007. Saint-Petersburs. Russia. 639 p. (In Russian).Баранов ВС, Кузнецова ТВ, Цитогенетика эмбрионального развития человека. 2007. СПб Изд-во Н-Л. 639 с.Vlahos A, Mansell T, Saffery R, Novakovic B. Human placental methylome in the interplay of adverse placental health, environmental exposure, and pregnancy outcome. PLoS Genet. 2019;15(8): e1008236. doi: 10.1371/journal.pgen.1008236Tolmacheva EN, Vasilyev SA, Nikitina TV, Lytkina ES, Sazhenova EA, Zhigalina DI, Vasilyeva OY, Markov AV, Demeneva VV, Tashireva LA, Kashevarova AA, Lebedev IN. Identification of differentially methylated genes in first-trimester placentas with trisomy 16. Sci Rep. 2022;12(1):1166. doi: 10.1038/s41598-021-04107-9Vasilyev SA, Tolmacheva EN, Vasilyeva OY, Markov AV, Zhigalina DI, Zatula LA, Lee VA, Serdyukova ES, Sazhenova EA, Nikitina TV, Kashevarova AA, Lebedev IN. LINE-1 retrotransposon methylation in chorionic villi of first trimester miscarriages with aneuploidy. J Assist Reprod Genet. 2021;38(1):139-149. doi: 10.1007/s10815-020-02003-1Lebedev IN, Ostroverkhova NV, Nikitina TV, Sukhanova NN, Nazarenko SA. Features of chromosomal abnormalities in spontaneous abortion cell culture failures detected by interphase FISH analysis. Eur J Hum Genet. 2004;12(7):513-20. doi: 10.1038/sj.ejhg.5201178Vasilyev SA, Timoshevsky VA, Lebedev IN. Cytogenetic mechanisms of aneuploidy in somatic cells of chemonuclear industry professionals with incorporated plutonium-239. Russian Journal of Genetics. 2010;46(11):1381-1385. doi: 10.1134/s1022795410110141Vasilyev SA, Tolmacheva EN, Kashevarova AA, Sazhenova EA, Lebedev IN. Methylation status of LINE-1 retrotransposon in chromosomal mosaicism during the early stages of human embryonic development. Molecular Biology (Moscow). 2015;49(1):144-152. doi: 10.1134/S0026893314060193. (In Russian).Васильев С.А., Толмачева Е.Н., Кашеварова А.А., Саженова Е.А., Лебедев И.Н. Статус метилирования ретротранспозона LINE-1 при хромосомном мозаицизме на ранних стадиях эмбрионального развития человека // Молекулярная биология. 2015. Т. 49. № 1. С. 165-174. doi: 10.7868/S0026898414060196V.Nishikata I, Sasaki H, Iga M, Tateno Y, Imayoshi S, Asou N, Nakamura T, Morishita K. A novel EVI1 gene family, MEL1, lacking a PR domain (MEL1S) is expressed mainly in t(1;3)(p36; q21)-positive AML and blocks G-CSF-induced myeloid differentiation. Blood. 2003;102(9):3323-32. doi: 10.1182/blood-2002-12-3944Mochizuki N, Shimizu S, Nagasawa T, Tanaka H, Taniwaki M, Yokota J, Morishita K. A novel gene, MEL1, mapped to 1p36.3 is highly homologous to the MDS1/EVI1 gene and is transcriptionally activated in t(1;3)(p36; q21)-positive leukemia cells. Blood. 2000;96(9):3209-14.Bjork B, Vieira A, Faust S, Camper S, Murray J, Beier D. Phenotypic, genetic, and developmental characterization of CPO1, a recessive ENU-induced mouse model of cleft palate. Mouse Molecular Genetics. Cold Spring Harbor Press, Woodbury, NY. 2006. 149 p.Kajimura S, Seale P, Tomaru T, Erdjument-Bromage H, Cooper MP, Ruas JL, Chin S, Tempst P, Lazar MA, Spiegelman BM. Regulation of the brown and white fat gene programs through a PRDM16/CtBP transcriptional complex. Genes Dev. 2008;22(10):1397-409. doi: 10.1101/gad.1666108Horn KH, Warner DR, Pisano M, Greene RM. PRDM16 expression in the developing mouse embryo. Acta Histochem. 2011;113(2):150-5. doi: 10.1016/j.acthis.2009.09.006Chen P, Piaggi P, Traurig M, Bogardus C, Knowler WC, Baier LJ, Hanson RL. Differential methylation of genes in individuals exposed to maternal diabetes in utero. Diabetologia. 2017; 60(4): 645-655. doi: 10.1007/s00125-016-4203-1Taschereau A, Thibeault K, Allard C, Juvinao-Quintero D, Perron P, Lutz SM, Bouchard L, Hivert MF. Maternal glycemia in pregnancy is longitudinally associated with blood DNAm variation at the FSD1L gene from birth to 5 years of age. Clin Epigenetics. 2023;15(1):107. doi: 10.1186/s13148-023-01524-7Cote S, Gagne-Ouellet V, Guay SP, Allard C, Houde AA, Perron P, Baillargeon JP, Gaudet D, Guerin R, Brisson D, Hivert MF, Bouchard L. PPARGC1alpha gene DNA methylation variations in human placenta mediate the link between maternal hyperglycemia and leptin levels in newborns. Clin Epigenetics. 2016;8:72. doi: 10.1186/s13148-016-0239-9Lim IY, Lin X, Teh AL, Wu Y, Chen L, He M, Chan SY, MacIsaac JL, Chan JKY, Tan KH, Chong MFF, Kobor MS, Godfrey KM, Meaney MJ, Lee YS, Eriksson JG, Gluckman PD, Chong YS, Karnani N. Dichotomy in the Impact of Elevated Maternal Glucose Levels on Neonatal Epigenome. J Clin Endocrinol Metab. 2022;107(3): e1277-e1292. doi: 10.1210/clinem/dgab710Wang WJ, Huang R, Zheng T, Du Q, Yang MN, Xu YJ, Liu X, Tao MY, He H, Fang F, Li F, Fan JG, Zhang J, Briollais L, Ouyang F, Luo ZC. Genome-Wide Placental Gene Methylations in Gestational Diabetes Mellitus, Fetal Growth and Metabolic Health Biomarkers in Cord Blood. Front Endocrinol (Lausanne). 2022;13:875180. doi: 10.3389/fendo.2022.875180Sugiyama T, Benitez CM, Ghodasara A, Liu L, McLean GW, Lee J, Blauwkamp TA, Nusse R, Wright CV, Gu G, Kim SK. Reconstituting pancreas development from purified progenitor cells reveals genes essential for islet differentiation. Proc Natl Acad Sci USA. 2013;110(31):12691-6. doi: 10.1073/pnas.1304507110Sasaki A, Murphy KE, Briollais L, McGowan PO, Matthews SG. DNA methylation profiles in the blood of newborn term infants born to mothers with obesity. PLoS One. 2022;17(5): e0267946. doi: 10.1371/journal.pone.0267946Seale P, Conroe HM, Estall J, Kajimura S, Frontini A, Ishibashi J, Cohen P, Cinti S, Spiegelman BM. Prdm16 determines the thermogenic program of subcutaneous white adipose tissue in mice. J Clin Invest. 2011;121(1):96-105. doi: 10.1172/JCI44271Benton MC, Johnstone A, Eccles D, Harmon B, Hayes MT, Lea RA, Griffiths L, Hoffman EP, Stubbs RS, Macartney-Coxson D. An analysis of DNA methylation in human adipose tissue reveals differential modification of obesity genes before and after gastric bypass and weight loss. Genome Biol. 2015;16(1):8. doi: 10.1186/s13059-014-0569-xSerrano A, Asnani-Kishnani M, Couturier C, Astier J, Palou A, Landrier JF, Ribot J, Bonet ML. DNA Methylation Changes are Associated with the Programming of White Adipose Tissue Browning Features by Resveratrol and Nicotinamide Riboside Neonatal Supplementations in Mice. Nutrients. 2020; 12(2). doi: 10.3390/nu12020461Willi-Monnerat S, Migliavacca E, Surdez D, Delorenzi M, Luthi-Carter R, Terskikh AV. Comprehensive spatiotemporal transcriptomic analyses of the ganglionic eminences demonstrate the uniqueness of its caudal subdivision. Mol Cell Neurosci. 2008;37(4):845-56. doi: 10.1016/j.mcn.2008.01.009Corley MJ, Vargas-Maya N, Pang APS, Lum-Jones A, Li D, Khadka V, Sultana R, Blanchard DC, Maunakea AK. Epigenetic Delay in the Neurodevelopmental Trajectory of DNA Methylation States in Autism Spectrum Disorders. Front Genet. 2019;10:907. doi: 10.3389/fgene.2019.00907Shull LC, Sen R, Menzel J, Goyama S, Kurokawa M, Artinger KB. The conserved and divergent roles of Prdm3 and Prdm16 in zebrafish and mouse craniofacial development. Dev Biol. 2020;461(2):132-144. doi: 10.1016/j.ydbio.2020.02.006Luedi PP, Dietrich FS, Weidman JR, Bosko JM, Jirtle RL, Hartemink AJ. Computational and experimental identification of novel human imprinted genes. Genome Res. 2007;17(12):1723-30. doi: 10.1101/gr.6584707Pi L, Zhang Z, Gu Y, Wang X, Wang J, Xu J, Liu J, Zhang X, Du J. DNA methylation profiling in recurrent miscarriage. Peer J. 2020;8: e8196. doi: 10.7717/peerj.8196Nishimura H, Ikawa Y, Kajikawa E, Shimizu-Mizuno N, Hiver S, Tabata-Okamoto N, Mori M, Kitajima T, Hayashi T, Yoshimura M, Umeda M, Nikaido I, Kurokawa M, Watanabe T, Hamada H. Maternal epigenetic factors in embryonic and postnatal development. Genes Cells. 2023;28(6):422-432. doi: 10.1111/gtc.13024Saenz-de-Juano MD, Ivanova E, Billooye K, Herta AC, Smitz J, Kelsey G, Anckaert E. Genome-wide assessment of DNA methylation in mouse oocytes reveals effects associated with in vitro growth, superovulation, and sexual maturity. Clin Epigenetics. 2019;11(1):197. doi: 10.1186/s13148-019-0794-yQin L, Luo X-Q, Wei J-X, Wei Y-X, Wang J-L, Zhang L-L, Huang H-B, Wei X-C. Genome-wide DNA methylation analysis of oligospermia and asthenozoospermia in a Chinese population. Int J Clin Exp Med. 2019;12(4):3168-3184.Pacheco SE, Houseman EA, Christensen BC, Marsit CJ, Kelsey KT, Sigman M, Boekelheide K. Integrative DNA methylation and gene expression analyses identify DNA packaging and epigenetic regulatory genes associated with low motility sperm. PLoS One. 2011;6(6): e20280. doi: 10.1371/journal.pone.0020280Roberts AL, Gladish N, Gatev E, Jones MJ, Chen Y, MacIsaac JL, Tworoger SS, Austin SB, Tanrikut C, Chavarro JE, Baccarelli AA, Kobor MS. Exposure to childhood abuse is associated with human sperm DNA methylation. Transl Psychiatry. 2018;8(1):194. doi: 10.1038/s41398-018-0252-1Murphy SK, Itchon-Ramos N, Visco Z, Huang Z, Grenier C, Schrott R, Acharya K, Boudreau MH, Price TM, Raburn DJ, Corcoran DL, Lucas JE, Mitchell JT, McClernon FJ, Cauley M, Hall BJ, Levin ED, Kollins SH. Cannabinoid exposure and altered DNA methylation in rat and human sperm. Epigenetics. 2018;13(12):1208- 1221. doi: 10.1080/15592294.2018.1554521Stimpfel M, Vrtacnik-Bokal E. Minor DNA methylation changes are observed in spermatozoa prepared using different protocols. Andrology. 2020; 8(5): 1312-1323. doi: 10.1111/andr.12832Gulino GM, Bruno F, Sturiale V, Brancato D, Ragusa D, Tosi S, Saccone S, Federico C. From FISH to Hi-C: The Chromatin Architecture of the Chromosomal Region 7q36.3, Frequently Rearranged in Leukemic Cells, Is Evolutionary Conserved. Int J Mol Sci. 2021;22(5). doi: 10.3390/ijms22052338Lan MS, Wasserfall C, Maclaren NK, Notkins AL. IA-2, a transmembrane protein of the protein tyrosine phosphatase family, is a major autoantigen in insulin-dependent diabetes mellitus. Proc Natl Acad Sci U S A. 1996;93(13):6367-70. doi: 10.1073/pnas.93.13.6367Lu J, Li Q, Xie H, Chen ZJ, Borovitskaya AE, Maclaren NK, Notkins AL, Lan MS. Identification of a second transmembrane protein tyrosine phosphatase, IA-2beta, as an autoantigen in insulin-dependent diabetes mellitus: precursor of the 37-kDa tryptic fragment. Proc Natl Acad Sci U S A. 1996;93(6):2307-11. doi: 10.1073/pnas.93.6.2307Wasmeier C, Hutton JC. Molecular cloning of phogrin, a protein-tyrosine phosphatase homologue localized to insulin secretory granule membranes. J Biol Chem. 1996;271(30):18161-70. doi: 10.1074/jbc.271.30.18161Dragic D, Ennour-Idrissi K, Michaud A, Chang SL, Durocher F, Diorio C. Association Between BMI and DNA Methylation in Blood or Normal Adult Breast Tissue: A Systematic Review. Anticancer Res. 2020;40(4):1797-1808. doi: 10.21873/anticanres.14134Ouni M, Saussenthaler S, Eichelmann F, Jahnert M, Stadion M, Wittenbecher C, Ronn T, Zellner L, Gottmann P, Ling C, Schulze MB, Schurmann A. Epigenetic Changes in Islets of Langerhans Preceding the Onset of Diabetes. Diabetes. 2020;69(11):2503-2517. doi: 10.2337/db20-0204Weng X, Liu F, Zhang H, Kan M, Wang T, Dong M, Liu Y. Genome-wide DNA methylation profiling in infants born to gestational diabetes mellitus. Diabetes Res Clin Pract. 2018;142:10-18. doi: 10.1016/j.diabres.2018.03.016Awamleh Z, Butcher DT, Hanley A, Retnakaran R, Haertle L, Haaf T, Hamilton J, Weksberg R. Exposure to Gestational Diabetes Mellitus (GDM) alters DNA methylation in placenta and fetal cord blood. Diabetes Res Clin Pract. 2021; 174: 108690. doi: 10.1016/j.diabres.2021.108690Yang IV, Zhang W, Davidson EJ, Fingerlin TE, Kechris K, Dabelea D. Epigenetic marks of in utero exposure to gestational diabetes and childhood adiposity outcomes: the EPOCH study. Diabet Med. 2018;35(5):612-620. doi: 10.1111/dme.13604Wang W, Yao W, Tan Q, Li S, Duan H, Tian X, Xu C, Zhang D. Identification of key DNA methylation changes on fasting plasma glucose: a genome-wide DNA methylation analysis in Chinese monozygotic twins. Diabetol Metab Syndr. 2023;15(1):159. doi: 10.1186/s13098-023-01136-4Li S, Wang W, Zhang D, Li W, Lund J, Kruse T, Mengel-From J, Christensen K, Tan Q. Differential regulation of the DNA methylome in adults born during the Great Chinese Famine in 1959-1961. Genomics. 2021;113(6):3907-3918. doi: 10.1016/j.ygeno.2021.09.018Lee S. The association of genetically controlled CpG methylation (cg158269415) of protein tyrosine phosphatase, receptor type N2 (PTPRN2) with childhood obesity. Sci Rep. 2019;9(1):4855. doi: 10.1038/s41598-019-40486-wRonn T, Perfilyev A, Jonsson J, Eriksson KF, Jorgensen SW, Brons C, Gillberg L, Vaag A, Stener-Victorin E, Ling C. Circulating triglycerides are associated with human adipose tissue DNA methylation of genes linked to metabolic disease. Hum Mol Genet. 2023;32(11):1875-1887. doi: 10.1093/hmg/ddad024Melton PE, Burton MA, Lillycrop KA, Godfrey KM, Rauschert S, Anderson D, Burdge GC, Mori TA, Beilin LJ, Ayonrinde OT, Craig JM, Olynyk JK, Holbrook JD, Pennell CE, Oddy WH, Moses EK, Adams LA, Huang RC. Differential DNA methylation of steatosis and non-alcoholic fatty liver disease in adolescence. Hepatol Int. 2023;17(3):584-594. doi: 10.1007/s12072-022-10469-7Yan Q, Forno E, Cardenas A, Qi C, Han YY, Acosta-Perez E, Kim S, Zhang R, Boutaoui N, Canino G, Vonk JM, Xu CJ, Chen W, Marsland A, Oken E, Gold DR, Koppelman GH, Celedon JC. Exposure to violence, chronic stress, nasal DNA methylation, and atopic asthma in children. Pediatr Pulmonol. 2021;56(7):1896-1905. doi: 10.1002/ ppul.25372Krishna RG, Vishnu Bhat B, Bobby Z, Papa D, Badhe B, Kalidoss VK, Karli S. Identification of differentially methylated candidate genes and their biological significance in IUGR neonates by methylation EPIC array. J Matern Fetal Neonatal Med. 2022; 35(3): 525-533. doi: 10.1080/14767058.2020.1727881Williams L, Seki Y, Delahaye F, Cheng A, Fuloria M, Hughes Einstein F, Charron MJ. DNA hypermethylation of CD3(+) T cells from cord blood of infants exposed to intrauterine growth restriction. Diabetologia. 2016;59(8):1714-23. doi: 10.1007/s00125-016-3983-7Garcia E, Wiemels J, Marconett C, Corona K, Howe C, Foley H, Lerner D, Lurvey N, Farzan S, Bastain T, Breton C, Mohazzab- Hosseinian S. Effect of Parental Adverse Childhood Experiences on Intergenerational DNA Methylation Signatures. Res Sq. 2023. doi: 10.21203/rs.3.rs-2977515/v1Sarkar S, Sujit KM, Singh V, Pandey R, Trivedi S, Singh K, Gupta G, Rajender S. Array-based DNA methylation profiling reveals peripheral blood differential methylation in male infertility. Fertil Steril. 2019;112(1):61-72 e1. doi: 10.1016/j.fertnstert.2019.03.020Agha G, Mendelson MM, Ward-Caviness CK, Joehanes R, Huan T, Gondalia R, Salfati E, Brody JA, Fiorito G, Bressler J, Chen BH, Ligthart S, Guarrera S, Colicino E, Just AC, Wahl S, Gieger C, Vandiver AR, Tanaka T, Hernandez DG, Pilling LC, Singleton AB, Sacerdote C, Krogh V, Panico S, Tumino R, Li Y, Zhang G, Stewart JD, Floyd JS, Wiggins KL, Rotter JI, Multhaup M, Bakulski K, Horvath S, Tsao PS, Absher DM, Vokonas P, Hirschhorn J, Fallin MD, Liu C, Bandinelli S, Boerwinkle E, Dehghan A, Schwartz JD, Psaty BM, Feinberg AP, Hou L, Ferrucci L, Sotoodehnia N, Matullo G, Peters A, Fornage M, Assimes TL, Whitsel EA, Levy D, Baccarelli AA. Blood Leukocyte DNA Methylation Predicts Risk of Future Myocardial Infarction and Coronary Heart Disease. Circulation. 2019;140(8):645- 657. doi: 10.1161/CIRCULATIONAHA.118.039357Krolevets M, Cate VT, Prochaska JH, Schulz A, Rapp S, Tenzer S, Andrade-Navarro MA, Horvath S, Niehrs C, Wild PS. DNA methylation and cardiovascular disease in humans: a systematic review and database of known CpG methylation sites. Clin Epigenetics. 2023;15(1):56. doi: 10.1186/s13148-023-01468-yvan Andel MM, Groenink M, van den Berg MP, Timmermans J, Scholte A, Mulder BJM, Zwinderman AH, de Waard V. Genome-wide methylation patterns in Marfan syndrome. Clin Epigenetics. 2021;13(1):217. doi: 10.1186/s13148-021-01204-4Chuang YH, Lu AT, Paul KC, Folle AD, Bronstein JM, Bordelon Y, Horvath S, Ritz B. Longitudinal Epigenome-Wide Methylation Study of Cognitive Decline and Motor Progression in Parkinson’s Disease. J Parkinsons Dis. 2019;9(2):389-400. doi: 10.3233/JPD-181549Grunblatt E, Mandel S, Jacob-Hirsch J, Zeligson S, Amariglo N, Rechavi G, Li J, Ravid R, Roggendorf W, Riederer P, Youdim MB. Gene expression profiling of parkinsonian substantia nigra pars compacta; alterations in ubiquitin-proteasome, heat shock protein, iron and oxidative stress regulated proteins, cell adhesion/cellular matrix and vesicle trafficking genes. J Neural Transm (Vienna). 2004;111(12):1543-73. doi: 10.1007/s00702-004-0212-1Sandor C, Robertson P, Lang C, Heger A, Booth H, Vowles J, Witty L, Bowden R, Hu M, Cowley SA, Wade-Martins R, Webber C. Transcriptomic profiling of purified patient-derived dopamine neurons identifies convergent perturbations and therapeutics for Parkinson’s disease. Hum Mol Genet. 2017;26(3):552-566. doi: 10.1093/hmg/ddw412Kochmanski J, Kuhn NC, Bernstein AI. Parkinson’s disease-associated, sex-specific changes in DNA methylation at PARK7 (DJ-1), SLC17A6 (VGLUT2), PTPRN2 (IA-2beta), and NR4A2 (NURR1) in cortical neurons. NPJ Parkinsons Dis. 2022;8(1):120. doi: 10.1038/s41531-022-00355-2Wang W, Li W, Wu Y, Tian X, Duan H, Li S, Tan Q, Zhang D. Genome-wide DNA methylation and gene expression analyses in monozygotic twins identify potential biomarkers of depression. Transl Psychiatry. 2021;11(1):416. doi: 10.1038/s41398-021-01536-yLiang F, Lv K, Wang Y, Yuan Y, Lu L, Feng Q, Jing X, Wang H, Liu C, Rayner S, Ling S, Chen H, Wan Y, Zhou W, He L, Wu B, Qu L, Chen S, Xiong J, Li Y. Personalized Epigenome Remodeling Under Biochemical and Psychological Changes During Long-Term Isolation Environment. Front Physiol. 2019;10:932. doi: 10.3389/fphys.2019.00932den Dekker HT, Burrows K, Felix JF, Salas LA, Nedeljkovic I, Yao J, Rifas-Shiman SL, Ruiz-Arenas C, Amin N, Bustamante M, DeMeo DL, Henderson AJ, Howe CG, Hivert MF, Ikram MA, de Jongste JC, Lahousse L, Mandaviya PR, van Meurs JB, Pinart M, Sharp GC, Stolk L, Uitterlinden AG, Anto JM, Litonjua AA, Breton CV, Brusselle GG, Sunyer J, Smith GD, Relton CL, Jaddoe VWV, Duijts L. Newborn DNA-methylation, childhood lung function, and the risks of asthma and COPD across the life course. Eur Respir J. 2019;53(4). doi: 10.1183/13993003.01795-2018Chen YC, Tsai YH, Wang CC, Liu SF, Chen TW, Fang WF, Lee CP, Hsu PY, Chao TY, Wu CC, Wei YF, Chang HC, Tsen CC Chang YP, Lin MC. Epigenome-wide association study on asthma and chronic obstructive pulmonary disease overlap reveals aberrant DNA methylations related to clinical phenotypes. Sci Rep. 2021;11(1):5022. doi: 10.1038/s41598-021-83185-1Izquierdo AG, Boughanem H, Diaz-Lagares A, Arranz-Salas I, Esteller M, Tinahones FJ, Casanueva FF, Macias-Gonzalez M, Crujeiras AB. DNA methylome in visceral adipose tissue can discriminate patients with and without colorectal cancer. Epigenetics. 2022;17(6):665-676. doi: 10.1080/15592294.2021.1950991