EPIGENETIC PATTERNS IN PLACENTAL PROGRAMMING OF PREECLAMPSIA Cindy M. Anderson, PhD, WHNP-BC, FAAN Michelle L. Wright, MS, RN Jody L. Ralph, PhD, RN Eric Uthus, PhD Joyce E. Ohm, PhD University of North Dakota College of Nursing School of Medicine and Health Sciences USDA ARS Grand Forks Human Nutrition Research Center

Introduction Preeclampsia o Pregnancy specific condition o De novo onset of hypertension and proteinuria in second half of pregnancy o Initiated in early pregnancy o Characterized by vascular dysfunction and placental insufficiency o Acute perinatal consequences 2

Introduction Preeclampsia o No reliable screening Diagnosis dependent on clinical manifestations of disease o Absence of definitive treatment Resolves after placenta delivery o Future risk for cardiovascular disease o Heritable risk for hypertension in offspring 3

Introduction Epigenetics o Study of heritable changes in gene expression o Modification of DNA without changes in DNA sequence o Above the gene DNA methylation Histone modification Micro RNA regulation 4

Introduction 5

Introduction 6

CpG Island Gene Promoter

CpG Island Gene Promoter S. Shore N. Shore

CpG Island Gene Promoter S. Shelf S. Shore N. Shelf N. Shore

Gene S. Shore S. Shelf N. Shelf N. Shore Transcription Factor Promoter CpG Island

Gene Promoter S. Shore S. Shelf N. Shelf N. Shore Transcription Factor CpG Island

GenePromoter S. Shore S. Shelf N. Shelf N. Shore Gene expression Transcription Factor CpG Island

Gene Promoter S. Shore S. Shelf N. Shelf N. Shore Gene S. Shore S. Shelf N. Shelf N. Shore Gene expression Transcription Factor CpG Island Promoter CpG Island

Gene S. Shore S. Shelf N. Shelf N. Shore Gene S. Shore S. Shelf N. Shelf N. Shore Gene expression Transcription Factor Promoter CpG Island Promoter CpG Island

Gene S. Shore S. Shelf N. Shelf N. Shore Gene S. Shore S. Shelf N. Shelf N. Shore Gene expression Transcription Factor Promoter CpG Island Promoter CpG Island

Gene S. Shore S. Shelf N. Shelf N. Shore Gene S. Shore S. Shelf N. Shelf N. Shore Gene expression Transcription Factor Gene expression repressed Transcription Factor Promoter CpG Island Promoter CpG Island

Gene S. Shore S. Shelf N. Shelf N. Shore Gene S. Shore S. Shelf N. Shelf N. Shore Gene expression Transcription Factor Gene expression repressed Transcription Factor Promoter CpG Island Promoter CpG Island

Gene S. Shore S. Shelf N. Shelf N. Shore Gene S. Shore S. Shelf N. Shelf N. Shore Gene expression Transcription Factor Gene expression repressed Transcription Factor Promoter CpG Island Promoter CpG Island

Central Hypothesis Distinct epigenetic patterns of DNA methylation are associated with heritable risk underlying preeclampsia

Methodology Design o Prospective Subjects o Nulliparous women recruited (n=55) o Maternal peripheral white blood cells (MPBCs) collected in first trimester of pregnancy o Placental tissue collection at delivery o Medical record abstraction for grouping

Methodology Genome-wide DNA methylation in women with normal pregnancy and preeclampsia o Maternal white blood cells o Placental chorionic tissue Method o Infinium Illumina bead array Analysis o Significant differences in methylation of individual CpG dinucleotides Beta score change of 0.2 o Functional classification DAVID v6.7 21

Differential methylation in maternal peripheral blood (delta-beta > 0.2; n=6/group) CpG dinucleotides ControlPE

CpG dinucleotides Mean differential methylation in maternal peripheral blood (delta-beta > 0.2; n=6/group) PreeclampsiaControl

Total Number of Differentially Methylated CpG Dinucleotides 25 Maternal Peripheral Blood

Gene location of significantly methylated CpG dinucleotides associated with preeclampsia 26 Maternal Peripheral Blood

Chromosomal Distribution of Differential Methylation 27 Chromosome

64% (n=216) 36% (n=125) Maternal Peripheral Blood Cells DNA Methylation Biomarkers of Preeclampsia from Mother In First Trimester of Pregnancy

64% 36% Transmission of DNA Methylation Biomarkers of Preeclampsia from Mother to Child during Pregnancy Placenta Maternal Peripheral Blood Cells

64% 36% Transmission of DNA Methylation Biomarkers of Preeclampsia from Mother to Child during Pregnancy Placenta Maternal Peripheral Blood Cells

Disease Categories Associated with Methylation Gain Cancer o Bladder o Breast o Colorectal o Endometrial o Head/neck Cardiovascular o Hypertension o Hypercholesterolemia Metabolic o Lipid Fatty acid cholesterol o Diabetes (type 1 and 2) o Obesity Neurologic o Chemical dependency o Anorexia o Bipolar disorder o Schizophrenia o Alzheimer’s disease Immune o Arthritis o Lupus erythematosus o Multiple sclerosis o Celiac disease o Thyroid autoimmunity

Disease Categories Associated with Methylation Loss Aging Cardiovascular o MI o Hypertension o Stroke o Hypercholesterolemia Metabolic o Lipid Fatty acid cholesterol o Diabetes (type 1 and 2) o Adiposity Neurologic o Chemical dependency o Bipolar disorder o Cognitive function o Asperger syndrome o Schizophrenia o Major depressive disorder o Alzheimer’s disease Immune o Arthritis o Asthma o Antiphospholipid syndrome o Grave’s disease o Hashimoto thyroiditis o Lupus erythematosus o Multiple sclerosis

Conclusions First study to identify common differentially methylated DNA o Early in pregnancies subsequently complicated by preeclampsia o Maternal and fetal origin Insight into mechanistic underpinnings of preeclampsia and familial risk Potential for early screening and targeted treatment 33

Acknowledgements Funding support o Robert Wood Johnson Nurse Faculty Scholar Award UND New Faculty Scholar Award o UND Research Experience for Medical Students (REMS) o Research team Jeanine Senti, MS, CNS 34

Thank you!