Placental identified NHIP regulating neuronal oxidative stress in autism

胎盘发现 NHIP 调节自闭症神经元氧化应激

• 批准号: 10717990
• 负责人: Janine M LaSalle
• 金额: $ 66.61万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10717990
• 关键词: 22q13; Adult; Affinity; Amino Acids; Animal Model; Antioxidants; Autism Diagnosis; Autopsy; Back; Binding; Binding Proteins; Biological Assay; Birth; Brain; CRISPR/Cas technology; Cell Hypoxia; Cell Line; Cell Nucleus; Cell physiology; Cells; Chromatin; Complex; Consumption; DNA Methylation; DNA-Binding Proteins; Data; Disease; Distal; Early Intervention; Embryo; Encapsulated; Energy consumption; Engineering; Enhancers; Environmental Risk Factor; Epigenetic Process; Etiology; Folic Acid Deficiency; Future; Genes; Genetic; Genetic Risk; Genetic Transcription; Genomics; Genotype; Goals; Human; Hypoxia; In Vitro; Maps; Messenger RNA; Metabolism; Methyl-CpG-Binding Protein 2; Methylation; Mitochondria; Modeling; Molecular; Multiprotein Complexes; Mus; Names; Neuronal Differentiation; Neuronal Hypoxia; Neurons; Oxidative Stress; Oxidative Stress Induction; Oxidative Stress Pathway; Oxygen; Oxygen Consumption; Pathway interactions; Peptides; Phenotype; Placenta; Placenta Diseases; Pre-Clinical Model; Pregnancy; Primates; Property; Proteins; Proteomics; Publishing; RNA Processing; RNA Splicing; Regulation; Reporter; Research; Respiration; Response Elements; Rett Syndrome; Sampling; Structure; Synapses; Testing; Therapeutic; Therapeutic Intervention; Transactivation; Transcript; Transcription Coactivator; Transcriptional Activation Domain; Translating; Translations; Valproic Acid; autism spectrum disorder; chromatin immunoprecipitation; cohort; comparison control; disorder risk; epigenome; epigenome-wide association studies; fetal; genetic risk factor; genomic locus; high risk; human fetal brain; human model; in utero; in vivo; in vivo Model; inhibitor; lipid nanoparticle; maternal obesity; mouse model; nanoparticle delivery; nerve stem cell; neurogenesis; neuroprotection; novel; overexpression; polygenic risk score; promoter; prospective; protein complex; protein protein interaction; resilience; response; risk variant; small molecule therapeutics; traffic-related air pollution; transcription factor

The human fetal brain consumes up to 60% of the body’s oxygen and energy consumption, despite making up ~13% of body mass. When the demand for oxygen in the placenta and developing brain exceeds its supply, hypoxia is induced, followed by changes to mitochondrial respiration, protein translation, and oxidative stress. Oxidative stress and epigenetic mechanisms within the placental-brain axis act at the interface of genetic and environmental risk factors in autism spectrum disorders. Using placental samples from a prospective high-risk cohort, we recently identified and named a novel gene NHIP (neuronal hypoxia inducible, placenta associated) and demonstrated its epigenetic, genetic, and transcriptional association with autism. NHIP is transiently expressed in response to hypoxia and neuronal differentiation, two examples of elevated oxidative stress. NHIP encodes a previously undiscovered micropeptide that localizes to the nucleus and is predicted to be neuroprotective, based on the lower expression of NHIP in placenta and brain samples from autism compared to control. The predicted structure of the NHIP peptide is an amphipathic helix that has similarity to a 9aaTAD motif found in transcriptional activation domains of many DNA binding proteins. We propose to test the hypothesis that NHIP acts as a competitive inhibitor of multi-protein complexes, thereby protecting developing and differentiating neurons following transient waves of hypoxia. Because NHIP is an “undiscovered protein” whose function had not been described before our recent study, this proposal will focus on the major research questions that are critical for determining the therapeutic relevance of NHIP. Specifically, what is the function of NHIP in neurons and brain, how is it regulated in response to hypoxia, and is it protective of neuronal oxidative stress? We propose three specific aims using well-characterized in vitro and in vivo models, including an inducible human neuronal cell line (LUHMES) engineered for NHIP transcript or peptide loss, human brain extracts with known NHIP genotype and expression levels, and mouse brain following NHIP peptide administration and/or hypoxia. Aim 1 will determine the molecular mechanisms of NHIP function and examine both protein-specific and global cellular impacts of NHIP loss. Aim 2 will determine how NHIP is transcriptionally responsive to hypoxia-induced oxidative stress by identifying the transcription factors and their genetic and epigenetic requirements for binding to the NHIP promoter and enhancer. Aim 3 will determine if exogenously delivered NHIP/NHIP protects neurons and embryonic neural precursor cells from hypoxia-induced oxidative stress. Together, the results from these proposed studies will provide the first functional characterization of NHIP, an understudied micropeptide that is associated with resilience to autism spectrum disorders. The potential impact of these results will be a potential therapeutic small molecule that could be used in early intervention therapy for autism and other neurodevelopmental or hypoxia-related disorders.

人胎儿的大脑消耗了全身高达60%的氧气和能量消耗， 尽管约占体重的 13%。当胎盘对氧气的需求量 发育中的大脑超出其供应，引起缺氧，随后线粒体发生变化 呼吸、蛋白质翻译和氧化应激。氧化应激和表观遗传机制 在胎盘-大脑轴内，在遗传和环境风险因素的界面上发挥作用 自闭症谱系障碍。使用来自前瞻性高风险队列的胎盘样本，我们 最近发现并命名了一个新基因NHIP（神经元缺氧诱导，胎盘 相关）并证明了其与自闭症的表观遗传、遗传和转录关联。 NHIP 因缺氧和神经元分化而短暂表达，两个例子 氧化应激升高。 NHIP 编码一种以前未被发现的微肽，该微肽定位于 基于 NHIP 在细胞核中的较低表达，预计具有神经保护作用 自闭症患者的胎盘和大脑样本与对照组的比较。 NHIP 的预测结构 肽是一种两亲性螺旋，与转录中发现的 9aaTAD 基序相似 许多 DNA 结合蛋白的激活域。我们建议检验 NHIP 的假设 作为多蛋白复合物的竞争性抑制剂，从而保护发育和 短暂缺氧波后分化神经元。因为 NHIP 是一个“未被发现的 在我们最近的研究之前，其功能尚未被描述过，该提案将重点关注 对确定治疗相关性至关重要的主要研究问题 国家健康保险计划。具体来说，NHIP在神经元和大脑中的功能是什么，它是如何被调节的？ 对缺氧的反应，它是否能保护神经元氧化应激？我们提出三项具体建议 目标是使用特征良好的体外和体内模型，包括可诱导的人类神经元 细胞系 (LUHMES) 专为 NHIP 转录物或肽丢失而设计，人脑提取物 已知 NHIP 基因型和表达水平，以及小鼠大脑追踪 NHIP 肽 给药和/或缺氧。目标 1 将确定 NHIP 功能的分子机制 并检查 NHIP 损失对蛋白质特异性和整体细胞的影响。目标 2 将决定 NHIP 如何通过识别对缺氧诱导的氧化应激进行转录反应 转录因子及其与 NHIP 结合的遗传和表观遗传要求 启动子和增强子。目标 3 将确定外源传递的 NHIP/NHIP 是否具有保护作用 神经元和胚胎神经前体细胞免受缺氧诱导的氧化应激。一起， 这些拟议研究的结果将提供 NHIP 的第一个功能特征， 一种尚未被研究的微肽，与自闭症谱系障碍的恢复能力有关。这 这些结果的潜在影响将是一种可以使用的潜在治疗小分子 用于自闭症和其他神经发育或缺氧相关的早期干预治疗 失调。