Prenatal Origins of Neurometabolic Consequences

神经代谢后果的产前起源

• 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
• 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' 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

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 (NDD) 的发病率不断上升。 NDD 的原因包括缺血性 4 种胎盘疾病 (IPD)，伴有胎儿/子宫内生长受限 (FGR/IUGR)、围产期窒息和 5.低血糖。人们已经开始探索异常胎盘健康与 NDD 之间的联系。除了 6 子宫内环境（IUE），基因突变也有助于 NDD 的子集，形成 IUE 基因 7范式。我们重点关注神经元葡萄糖转运蛋白亚型 3（GLUT3；基因：glut3）。 Glut3基因 据报道，8 种突变会导致 NDD/认知障碍。暴露于不良 IUE 可能导致 Glut3 突变 图 9 描绘了对 NDD 内表型的广泛影响。诊断学和早期饮食的发展 非常需要 10 次干预。我们已经证明，IUGR 和缺氧缺血会扰乱大脑的发育3 11 表达扰乱神经行为。我们还创建了小鼠 glut3 缺失，从而减少了跨胎盘 12 葡萄糖转运导致产后 NDD（兴奋性自闭症谱系障碍），其中细胞外小 13 种囊泡 (sEV) 为诊断提供动力，并且正在探索生酮饮食干预。接下来我们就脱离了 14 胎盘 glut3 基因从神经特异性 glut3 突变破译独立神经机制 落后 NDD 15。我们还利用诱导多能干细胞创建了表达 glut3 的人脑类器官 16（iPSC）。评估针对 NDD 的临床前生酮饮食效果将产生新的结果。为了实现这一目标 17 目标，我们将测试以下假设：IUE 和神经 glut3 突变/依赖性通过以下方式导致 NDD 18 扰乱神经发育，但有改善的潜力。目标是：1）a．调查变化 具有神经祖细胞的大脑皮层 (CC) 中每种细胞类型和细胞特异性转录组学的细胞数量为 19 使用巢蛋白驱动的条件无效出生后小鼠观察 20 细胞 (NPC) 中 glut3 的缺失。这将实现 21 x 10X 基因组学单细胞 (sc) RNA 测序和生物信息学分析，然后进行原位杂交 22 (ISH)/免疫组织化学 (IHC) 检测特定关键转录/翻译产物的主要变化 23 种细胞类型。 b.评估产前与产后生酮饮食作为早​​期干预的作用 24 改善NDD。 2) a.探索具有或不具有 MoMCR/IUGR 的 CC 中的神经过程和细胞特征 使用 Emx1 驱动的条件条件，在兴奋性神经元/NPC 中进行 25 个 glut3 的靶向缺失或过度表达 (OE) 26只裸鼠和OE小鼠从胚胎、出生后到成年的生命过程。这将需要去卷积 27 批量 CC RNA-seq 与 ISH/IHC，可非侵入性检测循环中受干扰的转录组/蛋白质 28 sEV。 b.研究对成人脑脊液/血浆代谢组学、神经元功能和神经行为的影响 29个后代。 3) a.从具有 glut3 OE 和/或 glut3 删除的对照 iPSC 中开发皮质类器官，以及 30 通过使用 scRNA-seq 和 ISH/IHC 解卷积类器官 RNA-seq 来检查细胞概况。 b.审问 31 缺氧和低血糖（有/无酮/乳酸）对 iPSC、NPC 和皮质类器官的影响。