Genomic imprinting is a molecular process that distinguishes the parental origins of certain genes, such that one allele is expressed and the opposite allele is silent. Many imprinted genes are expressed in the placenta, where some are known to play critical roles in placental development and metabolism. Most imprinted genes are organized in tightly linked clusters. There are at least 16 imprinted clusters in the mammalian genome, with the imprinting of each governed by a differentially methylated domain (DMD). One parental allele of a DMD becomes methylated during gametogenesis, and this methylation is perpetuated after fertilization by the action of the DNA cytosine methyltransferase 1 (DNMT1) enzyme. At the 8-celt embryonic stage this activity is from the maternal-effect isoform DNMTIo. Mouse embryos genetically engineered to lack oocyte-derived DNMTIo have 50% of the normal level of DMD methylation and are epigenetic mosaics of cells lacking DMD methylation on different imprinted clusters. Imprinting in the placenta is also severely disrupted in the absence of DNMTIo. Thus, the genetic model of DNMTIo deficiency is useful to study the role of genomic imprinting in placental function. The main objective of this research is to determine the global role of imprinting in the metabolic function of the mouse placenta by identifying the functional defects in DNMTIo-deficient placentas. This objective will be pursued in three Aims. A i m i : Define causal relationships between defects in imprinting and fuel metabolism in DNMTIodeficient placentas. The relationship among abnormalities in imprinted genes, morphology and metabolism in E9.5-E17.5 DNMTIo-deficient placentas will be determined using a combination of RNA in situ hybridization, histology and quantitative measurements of imprinted-gene expression and DNA methylation. We expect to find significant changes in the methylation of DMDs from one or more imprinted clusters and correlate these changes with specific morphological and metabolic defects.
Aim 2 : Identify novel pathways relevant to placental metabolism and function. The goal here is to use a variety of experimental techniques, including genome-wide expression studies, to identify established and novel metabolic pathways that are disrupted in DNMTIo-deficient placentas. We expect to find defects in the development of the maternal-fetal interface, lipid metabolism, and glycogen synthesis, breakdown and migration of glycogen producing cells.
Aim 3 : Determine the role of discrete imprinted genes in processing of placental metabolic fuels. Based on a number of criteria, the roles of individual imprinted genes in placental metabolism will be studied in knockout mice and transgenic lines overexpressing a gene to twice its normal level.

Public Health Relevance

An intact function of the human placenta is needed to supply nutrients and oxygen to the developing fetus, and therefore for successful outcomes of pregnancies. This research will investigate the role of a small but critically important set of imprinted genes in the normal formation and function of the placenta.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Program Projects (P01)
Project #
5P01HD069316-02
Application #
8486295
Study Section
Special Emphasis Panel (ZHD1-DSR-Z)
Project Start
Project End
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
2
Fiscal Year
2013
Total Cost
$224,453
Indirect Cost
$1,316
Name
Magee-Women's Research Institute and Foundation
Department
Type
DUNS #
119132785
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Ouyang, Yingshi; Bayer, Avraham; Chu, Tianjiao et al. (2016) Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity. Placenta 47:86-95
Mishima, Takuya; Sadovsky, Elena; Gegick, Margaret E et al. (2016) Determinants of effective lentivirus-driven microRNA expression in vivo. Sci Rep 6:33345
Lee, Sungeun; Pallerla, Srinivas R; Kim, Suyeon et al. (2016) Esrrb-Cre excises loxP-flanked alleles in early four-cell embryos. Genesis 54:53-61
Shaffer, Ben; McGraw, Serge; Xiao, Siyu C et al. (2015) The DNMT1 intrinsically disordered domain regulates genomic methylation during development. Genetics 199:533-41
Himes, K P; Young, A; Koppes, E et al. (2015) Loss of inherited genomic imprints in mice leads to severe disruption in placental lipid metabolism. Placenta 36:389-96
Koppes, Erik; Himes, Katherine P; Chaillet, J Richard (2015) Partial Loss of Genomic Imprinting Reveals Important Roles for Kcnq1 and Peg10 Imprinted Domains in Placental Development. PLoS One 10:e0135202
Larkin, Jacob; Chen, Baosheng; Shi, Xiao-Hua et al. (2014) NDRG1 deficiency attenuates fetal growth and the intrauterine response to hypoxic injury. Endocrinology 155:1099-106
Mohammadyani, Dariush; Tyurin, Vladimir A; O'Brien, Matthew et al. (2014) Molecular speciation and dynamics of oxidized triacylglycerols in lipid droplets: Mass spectrometry and coarse-grained simulations. Free Radic Biol Med 76:53-60
Makkar, A; Mishima, T; Chang, G et al. (2014) Fatty acid binding protein-4 is expressed in the mouse placental labyrinth, yet is dispensable for placental triglyceride accumulation and fetal growth. Placenta 35:802-7
Larkin, J C; Sears, S B; Sadovsky, Y (2014) The influence of ligand-activated LXR on primary human trophoblasts. Placenta 35:919-24