Risk Genes and Environmental Interactions in Neural Tube Defects

神经管缺陷的风险基因和环境相互作用

基本信息

批准号：
9357632
负责人：
MARGARET ELIZABETH ROSS
金额：
$ 134.64万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-23 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9357632
关键词：
3-Dimensional Affect Anencephaly and spina bifida X linked Arsenic Biological Birth CRISPR/Cas technology Cell Culture Techniques Cell Polarity Cells Child Complex Congenital Abnormality Cyst Data Data Set Defect Development Embryo Embryonic Development Ensure Environment Environmental Exposure Environmental Risk Factor Equilibrium Family Folic Acid Frequencies Funding Generations Genes Genetic Genetic Polymorphism Genetic study Goals Grant Human Human Genetics Hydrogen Peroxide In Vitro Individual Light Measures Mediating Metabolic Pathway Metabolism Methods Mitochondria Modeling Molecular Mus Mutant Strains Mice Mutation NOS3 gene Neural Tube Closure Neural Tube Defects Neural tube Neuroepithelial Neuroepithelial Cells Nitrates Nitric Oxide Synthase Nutritional status Outcome Oxidation-Reduction Oxidative Stress Parents Pathway interactions Patients Peroxonitrite Pharmaceutical Preparations Phosphorylation Population Prevention Prevention strategy Proteins Proteomics Regimen Risk Risk Assessment Role Serine Severities Shotguns Signal Pathway Source Spinal Dysraphism Stem cells Stress Structural defect Structure Superoxides Supplementation Technology Teratogens Testing Translations Untranslated RNA Variant base case control cell injury cohort embryo culture fetal folic acid supplementation gene environment interaction genetic risk factor genetic variant genome editing high throughput screening human embryonic stem cell human genomics human stem cells human subject humanized mouse improved metabolomics mouse model nerve stem cell neural model nitration nitrosative stress novel novel strategies prevent programs rare variant relating to nervous system risk variant screening small molecule stable isotope stem cell differentiation trafficking whole genome

项目摘要

RISK GENES AND ENVIRONMENT INTERACTIONS IN NTDS Neural tube defects (NTDs) arise from a complex interplay of multiple genes and environmental exposures. In human populations, folic acid (FA) supplementation can prevent up to 70% of NTD occurrences- -including anencephaly and spina bifida—by as yet unknown mechanism(s). Nevertheless, FA fails to benefit at least a third of families and recent data suggest that in some specific genetic contexts, FA may be deleterious to the developing embryo. Clearly, families would be far better served if their individual risks could be accurately assessed, including identification of which aspect of the FA metabolic pathway--or which supplement involving another pathway entirely--would provide the most benefit to them, so that NTD prevention strategies could be optimized according to individual genetic risk factors. This program aims to improve NTD risk assessment and prevention by integrating advanced human genomics with biological paradigms in humans and mice for identifying key gene-environment interactions. Project 1 (Ross PI with Finnell & Gross) has accumulated 200 whole genome sequences (WGS) from cases and 200 controls and has identified rare nonsense, frameshift and non-coding variants associated with spina bifida. In the renewal, we will employ a powerful high throughput method using molecular inversion probes (MIPs) to resequence a replication cohort of over 2,000 NTD cases. Cutting edge CRISPR-Cas9 dependent genome editing in hESCs and mice will probe the functional impact of identified variants on neuroepithelial cell polarity, proliferation, and the generation of reactive oxidative/nitrosative species (RONS). Project 2 (Gross PI with Ross & Finnell) will test the hypothesis that a major role for folate protection against NTD is to suppress the generation of RONS. They will employ a novel untargeted stable isotope method to trace folate-mediated 1-C trafficking in NTD-susceptible mouse models. In addition, they will employ a novel redoxome platform to quantify oxidatively-modified small molecules in NTD prone mice. With Projects 1&3, they will examine the impact of identified NTD associated human variants on cellular redox status and 1- C trafficking and the extent to which supplementation with small molecules can modulate these actions. Project 3 (Finnell PI with Gross & Ross) will examine the interaction of genetic variants and RONS to disrupt signaling pathways and cause cell damage during NT closure. They will test the ability of a human NTD-associated variant in NO synthase, NOS3, to increase ROS peroxynitrite in cells due to the phosphorylation of NOS3 on Ser633. It will test whether mitochondria are a major source of RONS during neurulation. Together, Projects 1, 2, & 3 will help define interactions of maternal/embryonic genetics, nutritional status and 1-C metabolism with NTD risk, using extensive human genomics, proteomics/metabolomics, and CRISPR-Cas9-dependent genome editing in hESCs, patient stem cells (iPSCs) and mice.

NTDS 的风险基因和环境相互作用神经管缺陷 (NTD) 是由多个基因和环境的复杂相互作用引起的在人群中，补充叶酸 (FA) 可以预防高达 70% 的 NTD 发生 - - 包括无脑畸形和脊柱裂——通过尚不清楚的机制然而，FA 未能受益。至少三分之一的家庭和最近的数据表明，在某些特定的遗传背景下，FA 可能是显然，如果家庭的个人风险能够得到改善，那么他们会得到更好的服务。进行准确评估，包括确定 FA 代谢途径的哪个方面，或者哪个方面完全涉及另一种途径的补充剂将为他们提供最大的利益，因此 NTD 可以根据个体遗传危险因素优化预防策略。该计划旨在通过整合先进的人类技术来改进 NTD 风险评估和预防基因组学与人类和小鼠的生物范例，用于识别关键的基因-环境相互作用。项目 1（Ross PI 与 Finnell & Gross）已积累了 200 个全基因组序列 (WGS) 病例和 200 个对照，并已识别出与在更新中，我们将采用一种强大的高通量方法，即分子反转。探针 (MIP) 对 2,000 多个 NTD 病例的复制队列进行重新测序。 hESC 和小鼠中的依赖性基因组编辑将探讨已识别变异对神经上皮细胞极性、增殖和活性氧化/亚硝基物质 (RONS) 的产生。项目 2（Ross & Finnell 的总 PI）将检验叶酸保护的主要作用这一假设对抗 NTD 的目的是抑制 RONS 的产生，他们将采用一种新型的非靶向稳定同位素。此外，他们还将采用在 NTD 易感小鼠模型中追踪叶酸介导的 1-C 贩运的方法。一种新颖的氧化还原体平台，用于量化 NTD 易感小鼠中的氧化修饰小分子。 1&3，他们将检查已识别的 NTD 相关人类变异对细胞氧化还原状态的影响和 1- C 运输以及补充小分子可以调节这些作用的程度。项目 3（Finnell PI 与 Gross & Ross）将检查遗传变异和 RONS 的相互作用，以他们将测试人类的能力。 NO 合酶 NOS3 中与 NTD 相关的变体，由于 NOS3 在 Ser633 上的磷酸化将测试线粒体是否是 RONS 的主要来源。项目 1、2 和 3 将共同帮助定义母体/胚胎遗传学、营养的相互作用。利用广泛的人类基因组学、蛋白质组学/代谢组学和 hESC、患者干细胞 (iPSC) 和小鼠中的 CRISPR-Cas9 依赖性基因组编辑。