Prenatal Origins of Neurometabolic Consequences

神经代谢后果的产前起源

基本信息

批准号：
10477429
负责人：
Sherin U Devaskar
金额：
$ 64.44万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10477429
关键词：
Adult Adult Children Affect Alzheimer&apos s Disease Anxiety Asphyxia Neonatorum Attention deficit hyperactivity disorder Behavioral Bioinformatics Biological Process Brain Brain Hypoxia-Ischemia CRISPR/Cas technology Caloric Restriction Cell Count Cells Cerebral cortex Child Clinical Copy Number Polymorphism Cortical Dysplasia DNA Sequence Alteration Dementia Dependence Detection Development Developmental Delay Disorders Diagnostic Dietary Intervention Early Intervention Embryo Environment Exposure to Failure Fetal Growth Retardation Fetus GLUT-3 protein Gene Deletion Gene Mutation Genes Genetic Transcription Genomics Germ Lines Gliosis Glucose Glucose Transporter Goals Health Human Huntington Disease Hyperactivity Hypoglycemia Hypoxia In Situ Hybridization Incidence Infant Injury Intervention Investigation Ketones Life Cycle Stages Light Link Malignant Neoplasms Meningomyelocele Mus Mutation Nerve Degeneration Neuraxis Neurodevelopmental Disorder Neurons Non-Invasive Cancer Detection Organoids Phenotype Placenta Diseases Plant Roots Plasma Process Protein Isoforms Proteins Reporting Rodent SLC2A1 gene Seizures Senile Plaques Societies Synapses Syndrome Testing Translating Vesicle autism spectrum disorder cell type chromosome 22q deletion syndrome cognitive disability diagnostic value dietary early screening endophenotype excitatory neuron exome extracellular extracellular vesicles fetal gene environment interaction genome sequencing glucose transport induced pluripotent stem cell infant outcome insight intrauterine environment ketogenic diet ketogentic metabolomics migration nerve stem cell nestin protein neurobehavior neurobehavioral neurodevelopment neurogenesis neuromechanism novel offspring overexpression postnatal pre-clinical premature prenatal relating to nervous system single-cell RNA sequencing transcriptome transcriptome sequencing transcriptomics whole genome

项目摘要

1 ABSTRACT 2 3 There is growing incidence of neurodevelopmental disorders (NDD). Causes for NDDs include ischemic 4 placental disorders (IPD) with fetal/intra-uterine growth restriction (FGR/IUGR), perinatal asphyxia, and 5 hypoglycemia. Exploration of connections between aberrant placental health and NDDs has occurred. Besides 6 the intra-uterine environment (IUE), genetic mutations also contribute to a subset of NDDs, forming an IUE-gene 7 paradigm. We have focused on the neuronal glucose transporter isoform 3 (GLUT3; gene: glut3). Glut3 gene 8 mutations are reported with NDDs/cognitive disabilities. Glut3 mutations with exposure to an adverse IUE may 9 portray expansive effects upon NDD endophenotypes. Development of diagnostics and early dietary 10 interventions is much needed. We have shown that IUGR and hypoxia-ischemia perturb developing brain glut3 11 expression perturbing neurobehavior. We also created murine glut3 deletions, that reduced trans-placental 12 glucose transport leading to postnatal NDD (excitatory autism spectrum disorders), where small extracellular 13 vesicles (sEVs) fueling diagnostics, and ketogenic dietary intervention are being explored. We next disengaged 14 placental glut3 gene from neural-specific glut3 mutations towards deciphering independent neural mechanisms 15 behind NDDs. We also created glut3 expressing human brain organoids from induced pluripotent stem cells 16 (iPSCs). Assessing pre-clinical ketogenic dietary effects targeting NDDs, will yield novel results. To achieve this 17 goal, we will test the hypothesis, that IUE and neural glut3 mutations/dependency cause NDDs by 18 perturbing neurodevelopment with a potential for amelioration. The aims are: 1) a. To investigate changes 19 in cell numbers per cell type and cell-specific transcriptomics in cerebral cortices (CC) with neural progenitor 20 cellular (NPC) absence of glut3 by using nestin-driven conditional null postnatal mice. This will be accomplished 21 by 10X genomics single cell (sc) RNA-sequencing and bioinformatic analyses, followed by in-situ hybridization 22 (ISH)/immunohistochemical (IHC) detection of major changes in key transcribed/translated products in specific 23 cell types. b. To assess administration of prenatal versus postnatal ketogenic diet as an early intervention in 24 ameliorating NDD. 2) a. To explore neural processes and cellular profile in CC with or without MoMCR/IUGR in 25 targeted absence or overexpression (OE) of glut3 in excitatory neurons/NPCs using Emx1-driven conditional 26 null and OE mice during the life course from embryonic and postnatal to the adult. This will entail deconvoluting 27 bulk CC RNA-seq with ISH/IHC, with non-invasive detection of perturbed transcriptome/proteins in circulating 28 sEVs. b. To examine the impact on CSF/plasma metabolomics, neuronal function and neurobehavior in the adult 29 offspring. 3) a. To develop cortical organoids from control iPSCs with glut3 OE and/or glut3 deletions, and 30 examine cellular profiles by deconvoluting organoid RNA-seq with scRNA-seq and ISH/IHC. b. To interrogate 31 the effect of hypoxia and hypoglycemia with/without ketones/lactate on iPSCs, NPCs and cortical organoids.

1摘要 2 3神经发育障碍的发生率越来越大（NDD）。 NDD的原因包括缺血 4个具有胎儿/鼻内生长限制（FGR/IUGR），围产期窒息和的胎盘疾病（IPD）和 5低血糖。发生了异常胎盘健康与NDD之间的联系。除了 6在河内环境（IUE），遗传突变也有助于NDD的子集，形成IUE-GENE 7范式。我们专注于神经元葡萄糖转运蛋白同工型3（GLUT3； Gene：Glut3）。 Glut3基因据报道8个突变患有NDD/认知障碍。暴露于不良IU的GLUT3突变可能 9描绘了对NDD内表型的广泛影响。诊断和早期饮食的发展非常需要10条干预措施。我们已经表明，IUGR和缺氧 - 缺血性扰动发育脑glut3 11表达神经行为。我们还创建了Murine Glut3删除，从而减少了跨置液 12导致出生后NDD（兴奋性自闭症谱系障碍）的葡萄糖转运，其中小细胞外正在探索13个囊泡（SEV）加油诊断和生酮饮食干预措施。我们接下来脱离了从神经特异性GLUT3突变到解密独立神经机制的14个胎盘GLUT3基因落后于NDD。我们还创建了从诱导的多能干细胞中表达人脑的glut3 16（IPSC）。评估针对NDD的临床前生酮饮食效应将产生新的结果。实现这一目标 17目标，我们将检验以下假设，即IUE和Neural Glut3突变/依赖性导致NDD 18扰动神经发育具有改善的潜力。目的是：1）a。调查变化 19在带有神经祖细胞的脑皮质（CC）中的细胞类型和细胞特异性转录组学的细胞数中 20个细胞（NPC）缺乏GLUT3，使用巢蛋白驱动的条件无效后小鼠。这将完成 21乘10倍基因组学单细胞（SC）RNA测序和生物信息学分析，然后是原位杂交 22（ISH）/免疫组织化学（IHC）检测主要转录/翻译产品的主要变化 23个细胞类型。 b。评估产前与产后生酮饮食的给药作为早期干预 24改善NDD。 2）a。探索CC中有或没有MOMCR/IUGR的神经过程和细胞剖面使用EMX1驱动的条件性的25个兴奋性神经元/NPC中GLUT3的靶向缺失或过表达（OE）在生命过程中，从胚胎和产后到成人的26个无效小鼠。这将需要切换 27大量CC RNA-Seq，带有ISH/IHC，在循环中无创的转录组/蛋白质侵入性检测 28 sevs。 b。检查成人中对CSF/血浆代谢组学，神经元功能和神经行为的影响 29后代。 3）a。从具有GLUT3 OE和/或GLUT3缺失的对照IPSC中开发皮质器官，以及 30通过用SCRNA-SEQ和ISH/IHC将器官RNA-seq解反应来检查细胞谱。 b。询问 31缺氧和低血糖伴有/不含酮/乳酸对IPSC，NPC和皮质器官的影响。