선천성 알콜노출에 의한 후세대 영향 및 기전연구(이봄이)

알코올의 태아에 미치는 영향에 관한 알코올의 태아에 미치는 영향에 관한 후성유전학적 기전 연구후성유전학적 기전 연구

유전학연구실 유전학연구실 이봄이이봄이

Cheil General Hospital & Women’s Healthcare CenterCheil General Hospital & Women’s Healthcare CenterLaboratory of Medical GeneticsLaboratory of Medical Genetics


• Studies on changes in gene expression that are

NOTNOT caused by changes in the sequence of DNA

• The mitotically and meiotically heritable control of

gene expression not related to DNA sequence

during development

Beyond the DNA Sequence!Beyond the DNA Sequence!

EpigenomeEpigenome


Heritable Heritable silencingsilencing

Interaction of heritable silencingInteraction of heritable silencing

•Small RNA mediated ControlSmall RNA mediated Control

•Post-translation modificationPost-translation modification •CpG islandCpG island

•Chromatin condensationChromatin condensation

Gene “switched OFF”Gene “switched OFF”• Silent (condensed) chromatin

• Methylated cytosines (red circles)

• Deacetylated histones

Gene “switched ON”Gene “switched ON”• Active (open) chromatin

• unmethylated cytosines (white circles)

• Acetylated histones

• The most heavily studied mode of epigenetic regulation

• By one of a variety of DNA methyltransferases (DNMTs) responsible for adding a methyl group to cytosine residues in cytosine/guanine- rich regions of DNA (called ‘CpG islands’)

CpG islandsCpG islands

• In and near approx. 40% of promoters of mammalian genes.

• About 70% of human promoters have a high CpG content.

• The “p” in CpG - phosphodiester bond between the cytosine and the guanine.


• Classic Teratogen affecting to reduce growth factor levels

- genetic, biochemical, cellular, and morphological

• Interferes

- DNA methylation

- histone modification

- noncoding RNA

• Dosage and timing – inducing target organ developmental

defect

During PregnancyDuring Pregnancy


• differences in timing and dosage underline this variability

• preconception, preimplantation, and gastrulation

• In Rat, placental mRNAs is altered after moderate drinking

during pregnancy

• Exposure to alcohol in the embryonic mouse can lead to

structural and neurophysiologic changes.

• Preconceptional effects mediated by paternal and maternal

alcohol consumption

Studies in the Animal ModelsStudies in the Animal Models


• General assessment about fetal development

• Limited human tissue specimen

• Partially researched on tissue specific epigenetic study

- Genome-wide methylation study via microarray

• Rarely challenged to elucidate the relation of epigenetic

changes within a family

Studies In Human Studies In Human

임신중 알코올 노출 임신부 및 태아 (n=100) vs

대조군 ( 임신중 알코올 비노출 임신부 및 태아 ) (n=100)

태아 알코올 증후군 및 알코올 대사 관련 대상유전자 선별

대상유전자 프로모터 부위 메틸화 양상 분석모체 : 노출 시기별 채혈

태아 : 출생 시 태반과 제대혈

태아 발달 평가임신중 태아 및 출생 후 발달 사항

자연유산 , 기형 , 조산 및 저체중증

기본정보수집 임신 초 , 중 , 말기 알코올 노출 기간 ,

빈도 , 음주량 , 기타 (흡연 , 약물 )

생체바이오마커 측정법 이용임신중 알코올 노출 시기별 모체의 알코올 노출력 평가

생체바이오마커 측정법 이용 출생 시 태변에서 FAEEs 측정

임신전 알코올 노출 배우자 및 태아 (n=100)vs

대조군 ( 임신전 알코올 비노출 배우자 및 태아 ) (n=100)

태아 알코올 증후군 및 알코올 대사 관련 대상유전자 선별

대상유전자 프로모터 부위 메틸화 양상 분석부체 : 노출 정도별 채혈

태아 : 출생 시 태반과 제대혈

태아 발달 평가임신중 태아 및 출생 후 발달 사항

자연유산 , 기형 , 조산 및 저체중증

기본정보수집 알코올 노출 기간 , 임신전 금주기간

빈도 , 음주량 , 기타 (흡연 , 약물 )

부체의 정액 검사정자수형태운동성

• Methylation pattern analysis - A genome-wide methylation analysis - micro array - Bisulfited genomic sequencing - Methylation-sensitive restriction enzyme - Gel electrophoresis analysis / fragment analysis

• Target Gene expression level analysis : - Quantitative Real-time PCR

• Candidate Target Gene : - Alcohol metabolism related genes ; ALDH2 (aldehyde dehydrogenase 2 family) , ADH1B (alcohol dehydrogenase 1B

(class I)

- Imprinting genes, Developmental related genes etc. ; H19 etc. - ADHD (attention-deficit hyperactivity disorder) related genes ; dopamine transporter gene, norepinephrine transporter gene

- DNA Methylation related regulating genes ; DNMT (DNA methyltransferase), MECP2 (methyl CpG-binding protein 2)

- Plasma homocysteine ; HERP (Homocysteine-induced ER protein)

methylationunmethylation

Restrictionenzyme

선천성 알콜노출에 의한 후세대 영향 및 기전연구(이봄이)

Health & Medicine