Ethanol-induced disruption of kinase signaling pathways in brain development

乙醇诱导大脑发育中激酶信号通路的破坏

基本信息

批准号：
10366867
负责人：
ERIC Christopher OLSON
金额：
$ 36.68万
依托单位：
UPSTATE MEDICAL UNIVERSITY
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：
2022-09-19 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10366867
关键词：
Actins Acute Adaptor Signaling Protein Alcohols Axon Biochemical Biological Brain Cells Centers for Disease Control and Prevention (U.S.)Child Critical Pathways Defect Dendrites Development Disease Dissection Dose Elements Embryo Enzymes Ethanol Ethanol dependence Etiology Event Fetal Alcohol Exposure Fetal Alcohol Syndrome Future Genetic Genetic Predisposition to Disease Genetic study Goals Golgi Apparatus Growth In Vitro Intellectual functioning disability Knock-out Ligand Binding Ligands Live Birth Location Membrane Methodology Molecular Molecular Target Morphology Multipolar Neuron Mus Nervous system structure Neuronal Differentiation Neurons Phase Phosphorylation Phosphotransferases Positioning Attribute Protein Dephosphorylation Protein Tyrosine Kinase Proteins Receptor Protein-Tyrosine Kinases Reelin Signaling Pathway Research Personnel Role Signal Pathway Signal Transduction Site Structure Synaptic plasticity Syndrome Teratogenic effects Testing Tyrosine Phosphorylation United States alcohol consumption during pregnancy alcohol effect alcohol exposure alcohol research alcohol response axon growth axonal pathfinding cofilin fetal glycosylation in vivo maternal alcohol use mind control novel postnatal prevent receptor receptor binding response src-Family Kinases synaptic function white matter

项目摘要

ABSTRACT Fetal Alcohol Syndrome (FAS) is one of the leading causes of intellectual disability in the United States. The CDC estimates that 0.2-1.5 per 1000 live births are children with FASD, a syndrome characterized by disrupted fetal brain development and postnatal intellectual disability (ID). Disrupted connectivity including altered dendritic structure, axonal pathfinding and white matter tracts are common findings in FAS and are thought to be major contributors to ID. However, the cellular and biological targets of alcohol are diverse and it is not clear whether there are common underlying molecular mechanisms producing these disruptions. Identification of common molecular mechanism(s) would enable a deeper understanding of this disorder, inform studies of genetic susceptibilities and provide molecular targets for neuroprotective strategies. This proposal pursues our finding that acute ethanol (EtOH) exposure disrupts Src kinase activity in embryonic cortical neurons. Src is a critical non-receptor tyrosine kinase that sits at central positions in multiple signaling pathways including the Reelin-Dab1 signaling pathway which controls brain layer formation and dendritogenesis. We found that acute EtOH exposure activates Src and induces phosphorylation of many proteins including Dab1, an essential adaptor protein in the Reelin-signaling pathway. Remarkably, this dramatic increase in phosphorylation is followed by a sustained dephosphorylation response in which the phosphorylation of Reelin effectors including Dab1, Src itself and the actin severing protein n-cofilin return to baseline levels, or below. During the extended dephosphorylation phase, the Reelin-signaling pathway can no longer be activated by in vitro application of its ligand, Reelin. In AIM 1 of this proposal, we will determine whether Reelin-Dab1 silencing occurs in vivo after maternal dosing with EtOH. We will then determine whether genetic deficiency in Src prevents the phosphorylation and dephosphorylation responses. Genetically establishing the critical kinase that initiates the EtOH response in vivo will be essential for future neuroprotective efforts. We and others have shown that Reelin-Dab1 signaling controls Golgi-deployment in the forming dendrite. In AIM 2 we will examine whether Src activation and inactivation disrupts Golgi location and function. Disrupted Golgi function would be expected to impact membrane addition, glycosylation, secretion and appropriate expression of many proteins, with potential long term negative consequences on neuritogenesis and neuronal function. In AIM 3 we will determine whether the EphA3 signaling pathway is similarly disrupted by Src dysregulation. EphA3 is a receptor tyrosine kinase that is required for axonal and white matter tract development. We identified the activation site of EphA3 as a target of EtOH-induced Src dysregulation raising the possibility that EphA3 activation and then silencing may contribute to FASD-related white matter disruptions. Collectively, these studies will determine the contribution of EtOH-dependent Src dysregulation to altered developmental signaling in pathways critical for brain development.

抽象的胎儿酒精综合征（FAS）是美国智力残疾的主要原因之一。这疾病预防控制中心估计，每1000个活产的0.2-1.5是患有FASD的儿童，一种以中断为特征的综合征胎儿脑发育和产后智力障碍（ID）。连通性中断，包括改变树突状结构，轴突探路和白质区是FAS中的常见发现，被认为是成为ID的主要贡献者。但是，酒精的细胞和生物学靶标是多种多样的，尚不清楚是否存在产生这些破坏的共同基本分子机制。识别常见的分子机制将使对这种疾病的了解更深入地了解遗传敏感性并为神经保护策略提供分子靶标。该提议追求我们发现急性乙醇（ETOH）暴露会破坏胚胎皮质神经元中的SRC激酶活性。 SRC是一个关键的非受体酪氨酸激酶，该激酶位于中心位置，以多种信号传导途径在内 Reelin-Dab1信号通路控制脑层的形成和树突生成。我们发现那个急性 ETOH暴露激活SRC并诱导许多蛋白质的磷酸化，包括DAB1，这是必不可少的 reelin信号途径中的衔接蛋白。值得注意的是，这种磷酸化的显着增加是然后是持续的去磷酸化反应，其中reelin效应子的磷酸化包括 DAB1，SRC本身和肌动蛋白切断的蛋白N-核蛋白恢复为基线水平或以下。在扩展期间去磷酸化阶段，不能再通过体外应用其激活reelin信号途径配体，reelin。在本提案的目标1中，我们将确定reelin-dab1沉默是否在体内发生用eTOH产妇给药。然后，我们将确定SRC的遗传缺乏是否阻止磷酸化和去磷酸化反应。基因建立启动的关键激酶 ETOH在体内的反应对于未来的神经保护作用至关重要。我们和其他人表明 reelin-dab1信号传导控制形成的树突中的高尔基人剥离。在AIM 2中，我们将检查SRC是否激活和灭活会破坏高尔基体的位置和功能。高尔基功能的破坏会有望影响膜添加，糖基化，分泌和适当表达许多蛋白质，对神经发生和神经元功能的潜在长期负面后果。在目标3中，我们将确定SRC失调是否类似地破坏EPHA3信号通路。 epha3是一个轴突和白质的发育所需的受体酪氨酸激酶。我们确定了 EPHA3的激活位点是EtOH诱导的SRC失调的靶标，这会引起Epha3的可能性激活然后沉默可能导致与FASD相关的白质中断。总的来说，这些研究将确定ETOH依赖性SRC失调对改变的发育信号传导的贡献在对大脑发育至关重要的途径中。