Mohammad Faisal J Khan1,2, Julian Little3, Valentina Aleotti1, Peter A Mossey4, Régine P M Steegers-Theunissen5,6, Luca Autelitano7, Maria C Meazzini7, Amin Ravaei1, Michele Rubini1. 1. Department of Biomedical and Specialty Surgical Sciences, Section of Medical Biochemistry, Molecular Biology and Genetics, University of Ferrara, Ferrara, Italy. 2. Epidemiology Research Group, Department of Public Health and Primary Care, Faculty of Medicine, KU Leuven - University, Leuven, Belgium. 3. School of Epidemiology and Public Health, University of Ottawa, Ottawa, Ontario, Canada. 4. Craniofacial Development at the World Health Organization-collaborating Centre for Oral and Craniofacial Research, Dental Hospital and School, University of Dundee, Dundee, UK. 5. Department of Obstetrics and Gynaecology, University Medical Center, Rotterdam, The Netherlands. 6. Division Neonatology Erasmus MC, Department of Pediatrics, University Medical Center, Rotterdam, The Netherlands. 7. Smile House, Regional Centre for Orofacial Clefts and Craniofacial Anomalies, Department of Cranio-Maxillo-Facial Surgery, San Paolo Hospital, University of Milan, Milan, Italy.
Abstract
OBJECTIVE: To investigate the influence of MTHFR c.677C>T genotype on LINE-1 methylation in lateral and medial tissues from cleft lip (CL). METHODS: Forty-five consecutive non-syndromic cleft lip with or without cleft palate (nsCL/P) cases were included in the study. Genomic DNA was extracted from tissues at both sides of cleft lip, and LINE-1 methylation was detected by bisulfite conversion and pyrosequencing. MTHFR c.677C>T genotyping was carried out using the TaqMan genotyping assay. RESULTS: LINE-1 methylation level was significantly higher on medial side of cleft lip compared with lateral side (p = 0.001). This difference was not significantly influenced by the case's sex or cleft type. However, MTHFR c.677C>T genotyping revealed that the difference in LINE-1 methylation across cleft lip was restricted to carriers of C allele of MTHFR c.677C>T and was not apparent in TT homozygous cases (p = 0.027). CONCLUSION: This integrated analysis supports the previous finding of differences in DNA methylation across the two sides of cleft lip and further suggests a possible role of MTHFR c.677C>T genotype in establishing this difference.
OBJECTIVE: To investigate the influence of MTHFR c.677C>T genotype on LINE-1 methylation in lateral and medial tissues from cleft lip (CL). METHODS: Forty-five consecutive non-syndromic cleft lip with or without cleft palate (nsCL/P) cases were included in the study. Genomic DNA was extracted from tissues at both sides of cleft lip, and LINE-1 methylation was detected by bisulfite conversion and pyrosequencing. MTHFR c.677C>T genotyping was carried out using the TaqMan genotyping assay. RESULTS: LINE-1 methylation level was significantly higher on medial side of cleft lip compared with lateral side (p = 0.001). This difference was not significantly influenced by the case's sex or cleft type. However, MTHFR c.677C>T genotyping revealed that the difference in LINE-1 methylation across cleft lip was restricted to carriers of C allele of MTHFR c.677C>T and was not apparent in TT homozygous cases (p = 0.027). CONCLUSION: This integrated analysis supports the previous finding of differences in DNA methylation across the two sides of cleft lip and further suggests a possible role of MTHFR c.677C>T genotype in establishing this difference.
Authors: Carlos Salamanca; Patricio González-Hormazábal; Andrea S Recabarren; Pamela A Recabarren; Roberto Pantoja; Noemi Leiva; Rosa Pardo; José Suazo Journal: Pediatr Res Date: 2020-06-03 Impact factor: 3.756