Cristiana Libardi Miranda Furtado1, Karina Bezerra Salomão2, Carolina Gennari Verruma2, Sarah Blima Paulino Leite2, Álvaro Fabrício Lopes Rios3, Monika Bialecka4, Ioannis Moustakas5, Hailiang Mei6, Claudia Cristina Paro de Paz7, Geraldo Duarte8, Susana M Chuva de Sousa Lopes4, Ester Silveira Ramos9. 1. Department of Genetics, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil; Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands; Department of Gynecology and Obstetrics, Ribeirão Preto Medical School, University of Sao Paulo, Sao Paulo, Brazil. 2. Department of Genetics, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil. 3. Biotechnology Laboratory, Center of Bioscience and Biotechnology, State University of North Fluminense Darcy Ribeiro, Campos dos Goitacazes, Rio de Janeiro, Brazil. 4. Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands. 5. Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands; Sequencing Analysis Support Core, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, the Netherlands. 6. Sequencing Analysis Support Core, Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, the Netherlands. 7. Department of Genetics, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil; Instituto de Zootecnia, Centro APTA de Bovinos de Corte, São Paulo, Brazil. 8. Department of Gynecology and Obstetrics, Ribeirão Preto Medical School, University of Sao Paulo, Sao Paulo, Brazil. 9. Department of Genetics, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil. Electronic address: esramos@usp.br.
Abstract
OBJECTIVE: To investigate the levels of DNA methylation in the KvDMR1 (KvLQT1 differentially methylated region 1) in embryonic and extra-embryonic tissues. DESIGN: Cross-sectional study. SETTING: University medical center and clinical hospital. PATIENT(S): Embryonic and/or extraembryonic tissues (umbilical cord, chorionic villus, chorion, decidua, and/or amnion) collected from 27 first-trimester pregnancies (up to 12 weeks of gestation, single embryos) from elective abortions, extravillous trophoblasts (EVTs) from the top of individual chorionic villi, and chorionic villi from 10 normal full-term placentas collected after birth. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): DNA methylation of the KvDMR1 region evaluated using quantitative analysis of DNA methylation followed by real-time polymerase chain reaction (qAMP) and bisulfite sequencing (bis-seq) analysis. RESULT(S): The results showed variability in KvDMR1 DNA methylation in different tissues from the same pregnancy. The average of DNA methylation was not different between the embryo, umbilical cord, amnion, and chorionic villi, despite the relatively low level of methylation observed in the amnion (33.50% ± 14.48%). Chorionic villi from term placentas showed a normal methylation pattern at KvDMR1 (42.60% ± 6.08%). The normal methylation pattern at KvDMR1 in chorionic villi (as well as in EVTs) from first-trimester placentas was confirmed by bis-seq. CONCLUSION(S): Our results highlight an existing heterogeneity in DNA methylation of the KvDMR1 region during first trimester and a consistent hypomethylation in the amnion in this period of gestation.
OBJECTIVE: To investigate the levels of DNA methylation in the KvDMR1 (KvLQT1 differentially methylated region 1) in embryonic and extra-embryonic tissues. DESIGN: Cross-sectional study. SETTING: University medical center and clinical hospital. PATIENT(S): Embryonic and/or extraembryonic tissues (umbilical cord, chorionic villus, chorion, decidua, and/or amnion) collected from 27 first-trimester pregnancies (up to 12 weeks of gestation, single embryos) from elective abortions, extravillous trophoblasts (EVTs) from the top of individual chorionic villi, and chorionic villi from 10 normal full-term placentas collected after birth. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): DNA methylation of the KvDMR1 region evaluated using quantitative analysis of DNA methylation followed by real-time polymerase chain reaction (qAMP) and bisulfite sequencing (bis-seq) analysis. RESULT(S): The results showed variability in KvDMR1 DNA methylation in different tissues from the same pregnancy. The average of DNA methylation was not different between the embryo, umbilical cord, amnion, and chorionic villi, despite the relatively low level of methylation observed in the amnion (33.50% ± 14.48%). Chorionic villi from term placentas showed a normal methylation pattern at KvDMR1 (42.60% ± 6.08%). The normal methylation pattern at KvDMR1 in chorionic villi (as well as in EVTs) from first-trimester placentas was confirmed by bis-seq. CONCLUSION(S): Our results highlight an existing heterogeneity in DNA methylation of the KvDMR1 region during first trimester and a consistent hypomethylation in the amnion in this period of gestation.
Authors: Herman Depypere; Yanrong Su; Nhi Dang; Bruce Poppe; Frank Stanczyk; Jaak Janssens; Jose Russo Journal: Eur J Cancer Prev Date: 2021-05-01 Impact factor: 2.164