Project 2: Prenatal Alcohol Exposure (PAE) and Fetal Alcohol Spectrum Disorder (FASD)

项目2：产前酒精暴露（PAE）和胎儿酒精谱系障碍（FASD）

• 批准号: 10540966
• 负责人: Kevin Peter Williams
• 金额: $ 18.65万
• 依托单位: NORTH CAROLINA CENTRAL UNIVERSITY
• 依托单位国家: 美国
• 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/10540966
• 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; Fetal alcohol effects; 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

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。这个目标 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 乙醇暴露。我们的方法利用转基因斑马鱼和小鼠，以及发育中的鳍和 分别为肢体，因为易于检查其分子信号传导途径和脆弱性的靶组织 乙醇致畸现象已有充分记录。本研究提案基于 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 PAE 的基本和新颖的见解 乙醇引起出生缺陷的机制。这一假设将通过实验进行检验，这些实验采用了良好的方法 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-环境相互作用，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。目标2是利用体外和体内高通量筛选技术来鉴定 挽救或加剧 PAE 影响的小分子调节剂。小分子化合物 挽救或加剧乙醇引起的斑马鱼胚胎鳍缺陷和小鼠四肢缺陷将 确定。拟议的研究将由国立政治大学的教师和实习生在 北卡罗来纳大学鲍尔斯酒精研究中心的研究人员。除了扩大我们对乙醇的了解之外 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。