Prenatal Alcohol Exposure and Fetal Alcohol Spectrum Disorder

产前酒精暴露和胎儿酒精谱系障碍

基本信息

批准号：
10541715
负责人：
Kevin Peter Williams
金额：
$ 2.74万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
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：
2022-09-20 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10541715
关键词：
Acetylcysteine Address Agonist Alcohols Animal Model Applications Grants Attenuated Behavioral Behavioral Model Biological Biological Markers Biological Models Candidate Disease Gene Cell Line Cell model Cells Chemicals Chick Cilia Clustered Regularly Interspaced Short Palindromic Repeats Cognitive deficits Congenital Abnormality Craniofacial Abnormalities Data Defect Development Digit structure Disease model Drug Screening Embryo Embryonic Development Ethanol Faculty Fetal Alcohol Exposure Fetal Alcohol Spectrum Disorder Fetal Development Fishes Fostering Foundations Future Genes Genetic Genetically Engineered Mouse Goals Growth Heterozygote Human In Vitro Incidence Intervention Knock-out Knockout Mice Libraries Limb Bud Limb Development Limb structure Literature Mentorship Mission Modeling Molecular Molecular Target Morphology Mus National Institute on Alcohol Abuse and Alcoholism Pathway interactions Pectoral Pharmacology Public Health Research Research Personnel Research Project Grants Research Proposals Resistance Role SHH gene Scientist Series Severities Signal Pathway Signal Transduction Small Interfering RNA Sonic Hedgehog Pathway Stains Technology Teratogenic effects Teratogens Testing Tissues Transgenic Organisms Tretinoin Underrepresented Minority Underrepresented Populations United States National Institutes of Health Visual Zebrafish alcohol effect alcohol exposure alcohol research alcohol sensitivity base beta Actin brain abnormalities clinical practice differential expression established cell line experimental study gene environment interaction high throughput screening in vitro Model in vivo in vivo Model innovation insight mouse genetics mouse model novel postnatal screening skeletal small hairpin RNA small molecule small molecule libraries smoothened signaling pathway teratogenesis transcriptome sequencing

项目摘要

Project Summary The objective of the proposed research is to target molecular pathways involved in alcohol-induced limb/fin defects to identify novel mechanisms with a role in FASD with the long-term goals of informing clinical practice, and ultimately reducing the incidence and severity of Fetal Alcohol Spectrum Disorders (FASD). This objective will be met by applying two innovative and complementary approaches that use cell, zebrafish and mouse models to (i) genetically knockout and identify genes that rescue or exacerbate the effects of ethanol exposure, and (ii) high throughput screening of focused chemical libraries to identify compounds that modulate the effects of ethanol exposure. Our approach utilizes genetically modified zebrafish and mice, and the developing fin and limb respectively, as readily examined target tissues whose molecular signaling pathways and vulnerability to ethanol teratogenesis are well documented. This research proposal is founded on 1) strong evidence that craniofacial abnormalities and limb defects have been observed in those identified with FASD, 2) recognition that disruption of sonic hedgehog (Shh) signaling, a major morphogenic pathway regulating embryonic development as a significant consequence of prenatal ethanol exposure (PAE), 3) that genetically reduced Shh signaling results in increased vulnerability to ethanol-induced birth defects, and 4) the availability of differentially expressed gene sets from our previous study on the effects of PAE on mouse limb development. The hypothesis to be tested is that the identification of novel genes and compounds that can rescue morphological and behavioral deficits elicited by PAE will reveal fundamental and novel insights into the mechanism of ethanol-induced birth defects. This hypothesis will be tested with experiments that employ well- established cell line, zebrafish, and mouse FASD models and will address the following Specific Aims: Aim 1 will use these in vivo and in vitro models to assess gene-environment interactions, particularly the candidate genes whose expression was shown to be altered in ethanol-exposed mouse limb buds by our previous ethanol RNA Seq study. Aim 2 is to utilize high-throughput screening technologies in vitro and in vivo to identify small molecule modulators that rescue or exacerbate the effects of PAE. Small molecule compounds that rescue or exacerbate ethanol-induced fin defects in zebrafish embryos and limbs defects in mice will be identified. The proposed studies will be conducted by NCCU faculty and trainees under the mentorship of UNC’s Bowles Center for Alcohol Studies researchers. In addition to expanding our understanding of ethanol’s teratogenic mechanism of action as well as providing clues regarding genetic sensitivities and interventions for FASD and potentially novel pathways with a role in FASD, this research project will foster state of the art research by budding scientists from underrepresented minorities and will provide the foundation for important FASD studies to be addressed in future grant applications/research.

项目概要拟议研究的目的是针对涉及酒精诱发肢体/鳍的分子途径缺陷以确定在 FASD 中起作用的新机制，其长期目标是为临床实践提供信息，最终降低胎儿酒精谱系障碍 (FASD) 的发生率和严重程度。将通过应用两种利用细胞、斑马鱼和小鼠的创新和互补方法来实现模型（i）基因敲除并识别可挽救或加剧乙醇暴露影响的基因， (ii) 对重点化学库进行高通量筛选，以识别调节效应的化合物我们的方法利用转基因斑马鱼和小鼠，以及正在发育的鳍和分别针对肢体，如容易检查的目标组织，其分子信号传导途径和脆弱性乙醇致畸的证据有据可查，该研究提案基于 1) 强有力的证据表明。在 FASD 患者中观察到颅面异常和肢体缺陷，2) 识别声音刺猬（Shh）信号的破坏，这是调节胚胎的主要形态发生途径发育是产前乙醇暴露 (PAE) 的一个重要后果，3) 会在遗传上降低 Shh 信号传导导致乙醇引起的出生缺陷的脆弱性增加，4) 我们之前关于 PAE 对小鼠肢体发育影响的研究中差异表达的基因集。要测试的假设是，识别可以拯救的新基因和化合物 PAE 引起的形态和行为缺陷将揭示对该假设将通过采用良好方法的实验进行检验。建立了细胞系、斑马鱼和小鼠 FASD 模型，并将实现以下具体目标：目标 1 将使用这些体内和体外模型来评估基因与环境的相互作用，特别是候选者我们之前的研究显示，暴露于乙醇的小鼠肢芽中表达发生改变的基因乙醇 RNA 测序研究的目标 2 是利用体外和体内高通量筛选技术来鉴定。小分子调节剂可挽救或加剧 PAE 的影响。拯救或恶化乙醇引起的斑马鱼胚胎鳍缺陷和小鼠四肢缺陷将是所提议的研究将由国立政治大学的教师和实习生在指导下进行。北卡罗来纳大学鲍尔斯酒精研究中心的研究人员不仅扩大了我们对乙醇的了解。致畸作用机制，并提供有关遗传敏感性和干预措施的线索 FASD 以及在 FASD 中发挥作用的潜在新颖途径，该研究项目将促进最先进的技术来自代表性不足的少数群体的崭露头角的科学家的研究将为重要的 FASD 研究将在未来的资助申请/研究中得到解决。