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Identifying Molecular Signatures of Genomic Imprinting Errors

识别基因组印记错误的分子特征

基本信息

批准号：
10445832
负责人：
Mellissa Rae Wigle Mann
金额：
$ 41.94万
依托单位：
MAGEE-WOMEN'S RES INST AND FOUNDATION
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10445832
关键词：
Address Affect Assisted Reproductive Technology Biological Biological Markers Biopsy Cells Chemicals Child Clinic Couples DNA Methylation Data Defect Development Disease Embryo Embryonic Development Epigenetic Process Fetal Growth Retardation Foundations Frequencies General Population Genes Genetic Genomic Imprinting Germ Cells Human Impairment Importins Incidence Infertility Knowledge Lead Low Birth Weight Infant Maintenance Mammals Mediating Medical Methods Methylation Modality Molecular Profiling Mus Mutagenesis North America Nuclear Import Oocytes Oogenesis Outcome Pathway interactions Pharmacology Population Prader-Willi Syndrome Pre-implantation Embryo Development Predisposition Premature Birth Procedures Proteins Proxy Publishing Regulation Reporter Reporting Research Proposals Risk Screening procedure Silver Silver-Russell syndrome Superovulation TRIM Gene Testing Time Transcript Unsafe Sex bisulfite blastocyst clinical practice embryo cell embryo culture imprint infertility treatment nucleocytoplasmic transport predictive marker predictive test preimplantation protein degradation single-cell RNA sequencing time use transcriptome sequencing trophoblast trying to conceive whole genome

项目摘要

Abstract Alarmingly, ~50 million couples worldwide are unable to conceive after 5 years of unprotected sex. Medically assisted reproductive technologies (ARTs) constitute important treatment modalities for infertile couples trying to conceive. While generally considered safe, ARTs are associated with higher incidences of preterm birth, intrauterine growth restriction, and low birth weight. Moreover, there are more Beckwith-Wiedemann, Silver- Russell, Angelman and Prader-Willi Syndrome children in the ART population harboring imprinted methylation errors compared to those in the general population. Since ART usage coincides with important stages of imprinted DNA methylation programming during gamete and preimplantation development, it is possible that ARTs lead to imprinting errors. Studies in fertile mice support this. Superovulation during oogenesis and/or embryo culture during preimplantation, two nearly universal ART procedures, lead to significant numbers of embryos with imprinted methylation errors. To date, little is known about the regulation of imprinted methylation maintenance during preimplantation development, and moreover, how ARTs lead to disruption in this maintenance. This proposal will address these knowledge gaps. In Aim 1, we will test the hypothesize that embryos are predisposed to imprinted methylation errors because ARTs disrupt crucial maternal-effect transcripts in oocytes that are required in embryos to maintain imprinted methylation. Candidate maternal- effect transcripts will be identified using single cell RNA-seq on control and superovulated oocyte, 1-cell and 2- cell embryos, representing the period of oocyte-to-embryo transition. Candidate function will then be determined though protein degradation methods. Next, we will determine whether altered maternal-effect transcripts in polar bodies, as a proxy for oocytes, correlates with imprinted methylation errors in resulting embryos, ultimately leading to identification of transcript biomarkers that are predictive of imprinted methylation errors. Aim 2 will test the importance of nuclear import in imprint maintenance using pharmacological and protein degradation methods, and whether it is disrupted by ARTs. In Aim 3, we hypothesize that the 4 to 8-cell stages represent a window of susceptibility when fast developing ART embryos acquire DNA methylation errors, particularly in outer, trophoblast cells. We will determine when embryo development is altered by ARTs using time-lapse analysis, and then compare DNA methylation perturbation in slow and fast developing embryos using whole genome bisulfite mutagenesis. This will distill a molecular signature of DNA methylation errors. Next, we will investigate this molecular signature during early development to determine when DNA methylation errors arise and in which lineages of ART embryos, as well as in cells induced to trophoblast fate. Finally, we will test the predictive power of developmental and morphokinetic parameters as a noninvasive procedure for embryos with a molecular signature of imprinted methylation errors. Results from this proposal will provide the biological basis for improvements to fertility treatments aimed at identifying at-risk embryos.

摘要