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.

项目摘要 这项研究的目的是针对酒精诱导的肢体/鳍相关的分子通路 缺陷以确定在FASD中起作用的新机制，并以告知临床实践为长期目标， 并最终降低胎儿酒精谱系障碍(FASD)的发生率和严重程度。这一目标 将通过应用使用细胞、斑马鱼和老鼠的两种创新和互补的方法来满足 (I)从基因上敲除和识别挽救或加剧酒精暴露影响的基因的模型， 和(2)高通量筛选重点化学库，以确定调节作用的化合物 暴露在酒精中。我们的方法利用了转基因斑马鱼和小鼠，以及发育中的鳍和 肢体分别作为易于检测的靶组织，其分子信号通路和易感性 乙醇的致畸作用是有据可查的。这项研究建议建立在1)强有力的证据基础上： 在FASD确诊的患者中观察到了颅面畸形和肢体缺陷，2)识别 Sonic Hedgehog(Shh)信号的中断，这是调节胚胎发育的主要形态发生途径 作为产前酒精暴露(PAE)的显著后果的发育，3)遗传减少 Shh信号导致对酒精引起的出生缺陷的易感性增加，以及4)可获得 与我们之前关于PAE对小鼠肢体发育影响的研究的差异表达基因集。 需要检验的假设是，新基因和化合物的鉴定可以挽救 PAE引起的形态和行为缺陷将揭示对 酒精致出生缺陷的机制。这一假设将通过实验进行验证，这些实验采用了良好的 建立了细胞系、斑马鱼和小鼠FASD模型，并将解决以下具体目标：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 我们先前的研究表明，乙醇暴露的小鼠肢芽中表达发生变化的基因 乙醇RNA序列研究。目标2是利用体外和体内的高通量筛选技术来鉴定 小分子调节剂，可挽救或加重PAE的影响。小分子化合物 挽救或加重乙醇诱导的斑马鱼胚胎鳍缺陷和小鼠肢体缺陷 确认身份。建议的研究将由NCCU的教职员工和实习生在 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理解 致畸作用机制以及提供有关遗传敏感性和干预的线索 FASD和潜在的在FASD中起作用的新途径，这项研究项目将促进最新技术的发展 来自少数族裔的初出茅庐的科学家的研究，将为重要的 FASD研究将在未来的赠款申请/研究中解决。