IMPACT OF LOSS OF THE AUTISM RISK GENE CHD8 ON EXPRESSION, EPIGENETICS AND CELLULAR FUNCTIONING OF THE DOPAMINERGIC SYSTEM

自闭症风险基因 CHD8 缺失对多巴胺能系统表达、表观遗传学和细胞功能的影响

• 批准号: 10660565
• 负责人: Rebecca Ann Muhle
• 金额: $ 69.33万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-17 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660565
• 关键词: Affect; Animal Model; Atlases; Attenuated; Basal Ganglia; Behavior; Behavioral; Binding; Biological; Biological Assay; Biology; Biotinylation; Brain; Brain Diseases; Cell physiology; Cells; Cellular biology; ChIP-seq; Child; Chromatin; Chronic; Collaborations; Complex; Corpus striatum structure; Cre driver; Critical Pathways; DNA; DNA Sequence Alteration; DNA-Binding Proteins; Data; Data Set; Defect; Development; Doctor of Medicine; Doctor of Philosophy; Dopamine; Dopamine Agonists; Dopamine Receptor; Electrophysiology (science); Epigenetic Process; Fellowship; Female; Fiber; Foundations; Funding; Future; Gene Expression; Genes; Genetic Risk; Genetic Transcription; Genomic Segment; Genomics; Goals; Human; Investigation; Knowledge; Label; Lifestyle-related condition; LoxP-flanked allele; Maps; Mentorship; Microdialysis; Midbrain structure; Modeling; Molecular; Molecular Abnormality; Molecular Genetics; Morbidity - disease rate; Mus; Mutation; National Institute of Mental Health; Neurobiology; Neurodevelopmental Disorder; Neurons; Nuclear; Nuclear RNA; Outcome; Pathway interactions; Pharmaceutical Preparations; Photometry; Play; Population; Psychiatrist; Public Health; Regulation; Research; Research Personnel; Resolution; Risk; Risk Factors; Role; Societies; Specific qualifier value; Stimulus; Structure; Symptoms; Synapses; System; Transgenes; Transposase; Universities; autism spectrum disorder; behavioral impairment; cell type; chromatin immunoprecipitation; chromatin remodeling; dopamine system; dopamine transporter; dopaminergic neuron; epigenetic profiling; extracellular; functional genomics; helicase; in vivo; insight; male; mortality; mouse model; multiple omics; neurobehavioral disorder; neurotransmission; novel; novel therapeutics; optogenetics; postsynaptic; presynaptic; promoter; repetitive behavior; response; risk variant; sex; social; stereotypy; tool; trait; transcriptome sequencing; transcriptomic profiling; transcriptomics; translational study; transmission process

PROJECT SUMMARY Dr. Rebecca Muhle is a practicing child psychiatrist and an Early Stage Investigator with a strong track record of developing molecular genetic tools to elucidate the mechanism of neurodevelopmental disorders. Her long- term goal is to identify cell types, circuits and regulatory networks affected by loss of Chromodomain Helicase DNA-binding Protein 8 (CHD8), a high confidence autism spectrum disorder (ASD) risk gene. Dr. Muhle initiated her studies of CHD8 during her fellowship with James Noonan, Ph.D. with co-mentorship by Matthew State, M.D., Ph.D. She successfully obtained research funding from the National Institute for Mental Health and the Simons Foundation for Autism Research to determine the impact of CHD8 and its loss on regulatory networks of the developing cortex, and she showed that CHD8 in developing cortex regulates other ASD risk genes. She is extending this research at Columbia University in collaboration with Jeremy Veenstra-VanderWeele, M.D. and Muhammed Osman Chohan, M.D., experts in translational animal models of neurobehavioral disorders and the study of monoaminergic neurotransmission. Together, they have found reduced stereotypy and attenuated hyperlocomotor activity in male, but not female Chd8+/- mice after inhibition of the dopamine transporter, with an intact response to dopamine receptor agonism, suggesting that CHD8 plays a critical role in the development of the dopaminergic circuit between the midbrain and striatum. The overall objective of this R01 proposal is to elucidate the mechanisms whereby CHD8 genetic mutations affect cells and pathways critical for core symptoms of ASD and other neurodevelopmental disorders, by integrating state-of-the-art single nuclear transcriptomics and cell type specific functional genomics with electrophysiologic and behavioral probing of animal models with a loss of CHD8 expression. The following three independent specific aims will address the central hypothesis that CHD8 regulates genes that are critical for the function of the basal ganglia and dopaminergic system, and that loss of CHD8 will dysregulate these genes and other ASD risk genes. This project will 1) identify the cellular, transcriptional, and epigenetic changes in the developing Chd8+/- mouse ventral midbrain and striatum using single nuclear RNA sequencing (snRNAseq) and chromatin accessibility (snATACseq), 2) elucidate how CHD8 loss causes a defect in the molecular transmission of the DA stimulus in the basal ganglia using fiber photometry and in vivo extracellular recordings using constitutive and conditional CHD8 haploinsufficiency mouse models, and 3) localize CHD8 binding activity in specific cell types of the dopaminergic system using chromatin immunoprecipitation and sequencing of a conditionally biotinylated Chd8-tag transgene. By studying CHD8, a key regulatory molecule that targets other ASD risk genes, Dr. Muhle will close a critical knowledge gap in the role of CHD8 in the circuitry implicated in ASD-related behaviors.

项目摘要 丽贝卡·穆尔（Rebecca Muhle）博士是一位执业的儿童精神病医生，也是一名具有良好往绩记录的早期舞台调查员 开发分子遗传工具来阐明神经发育障碍的机制。她的长期 术语目标是识别受染色体构酶损失影响的细胞类型，电路和调节网络 DNA结合蛋白8（CHD8），一种高置信自闭症谱系障碍（ASD）风险基因。 Muhle博士发起了 她在与James Noonan博士学位的研究期间对CHD8的研究。在马修州立大学的统治下， 医学博士她成功获得了美国国家心理健康研究所的研究资金和 Simons自闭症研究基金会确定CHD8及其对监管网络的损失的影响 在发育中的皮层中，她表明CHD8在开发皮层中调节其他ASD风险基因。 她正在哥伦比亚大学与Jeremy Veenstra-Vanderweele合作扩展这项研究， M.D.和Muhammed Osman Chohan，M.D。，神经行为疾病转化动物模型的专家 以及单胺能神经传递的研究。他们一起发现了刻板印象和 抑制多巴胺后，男性而非雌性CHD8 +/-小鼠的衰减超子运动活性 转运蛋白，对多巴胺受体激动剂的完整反应，表明CHD8在 中脑和纹状体之间多巴胺能回路的发展。此R01的总体目标 建议是阐明CHD8基因突变影响细胞和途径至关重要的机制 通过整合最新的单核，ASD和其他神经发育障碍的核心症状 转录组学和细胞类型特异性功能基因组学，具有电生理和行为探测 动物模型失去CHD8表达。 以下三个独立的特定目标将解决CHD8调节基因的中心假设 这对于基底神经节和多巴胺能系统的功能至关重要，而CHD8的丧失将 这些基因和其他ASD风险基因失调。该项目将1）确定细胞，转录和 使用单个核RNA的发育中的CHD8 +/-小鼠腹中脑和纹状体的表观遗传变化 测序（SNRNASEQ）和染色质可及性（SNATACSEQ），2）阐明CHD8损失如何导致缺陷 在使用纤维光度法和体内的基底神经节中DA刺激的分子传播中 使用本构和有条件的CHD8单倍宽度鼠标模型的细胞外记录，3） 使用染色质将CHD8结合活性定位在多巴胺能系统的特定细胞类型中 有条件生物素化的CHD8-TAG转基因的免疫沉淀和测序。通过研究CHD8，一个 靶向其他ASD风险基因的关键调节分子，Muhle博士将缩小有关临界知识差距 CHD8在与ASD相关行为有关的电路中的作用。