The molecular roles of RFX3 in neurodevelopment and Autism Spectrum disorder

RFX3 在神经发育和自闭症谱系障碍中的分子作用

基本信息

批准号：
10685268
负责人：
Jenny Lai
金额：
$ 5.27万
依托单位：
HARVARD MEDICAL SCHOOL
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：
2022-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10685268
关键词：
Address Affect Attention deficit hyperactivity disorder Autopsy Binding Binding Sites Brain Brain Diseases Cancer cell line Cell Culture Techniques Cell Nucleus Cells ChIP-seq Chromatin Clinical Communication Computer Analysis Data Defect Development Disease model Electrophysiology (science)Equilibrium Excitatory Synapse Genes Genetic Genetic Transcription Genomics Genotype Human Immunohistochemistry Impairment Individual Induced pluripotent stem cell derived neurons Knock-out Libraries Long-Term Depression Long-Term Potentiation Modeling Molecular Mus Mutate Neurobiology Neurodevelopmental Disorder Neuroglia Neuronal Differentiation Neurons Organoids Pathogenesis Pathway interactions Pattern Preparation Process Prosencephalon RFX3 Repression Research Research Proposals Risk Role Signal Transduction Social Interaction Specific qualifier value Statistical Data Interpretation Synapses Synaptic Transmission Synaptic plasticity Testing Time Tissues Training Variant autism spectrum disorder cell type ciliopathy cilium biogenesis cohort conditional knockout de novo mutation differential expression disorder risk excitatory neuron exome sequencing fetal genome sequencing genome-wide genome-wide analysis hippocampal pyramidal neuron human data human disease improved individuals with autism spectrum disorder induced pluripotent stem cell insight loss of function multi-electrode arrays nerve stem cell neural neurobiological mechanism neurodevelopment neuron development new therapeutic target next generation novel postnatal human programs promoter risk variant single-cell RNA sequencing skills synaptic function synaptogenesis transcription factor transcriptome transcriptome sequencing whole genome

项目摘要

Project Abstract Autism Spectrum Disorder (ASD) is the most common neurodevelopmental disorder, yet the neurobiological mechanisms underlying ASD pathogenesis remain largely unknown. Large-scale exome sequencing studies of individuals with ASD have identified over 100 genes significantly associated with ASD risk. Functional characterization of ASD risk genes can provide insight to ASD pathogenesis. We and others have recently identified de novo loss-of-function variants in the transcription factor RFX3 as a relatively common monogenic cause of ASD, implying an important role for RFX3 in human neurodevelopment. We have found evidence that RFX3 may be a critical transcriptional regulator of the development and function of layer II/III neurons: its expression is significantly enriched in cortical layer II/III excitatory neurons, and the RFX3 binding motif is specifically enriched in accessible chromatin regions of the human fetal germinal zone and layer II/III excitatory neurons. In this proposed research, I will address the hypothesis that RFX3 regulates key neurodevelopmental processes in layer II/III excitatory neurons and the expression of other ASD risk genes that affect neuronal formation and function. In Aim 1, I will identify the genes and pathways regulated by RFX3 in human cortical neurons by profiling the genome-wide binding sites of RFX3 and the transcriptional changes induced by loss of RFX3 occupancy in RFX3 haploinsufficient human iPSC-derived neurons. In Aim 2, I will evaluate the effect of RFX3 haploinsufficiency on cortical neuron formation and synaptic function in human iPSC-derived forebrain organoids. I will use single-cell RNA-sequencing to identify changes in cell type composition and infer alterations in developmental trajectories in RFX3 deficient organoids, and multielectrode array to assess synaptic plasticity balance in RFX3 deficient organoids compared to isogenic controls. Taken together, this proposal will yield insight on the transcriptional programs regulated by RFX3 in human neurons, and how RFX3 haploinsufficiency disrupts neuronal development and function. This will allow for improved understanding of ASD neurobiology, and the development of novel targeted therapies for ASD.

项目摘要自闭症谱系障碍 (ASD) 是最常见的神经发育障碍，但 ASD 发病机制的神经生物学机制仍然很大程度上未知。大规模对自闭症患者的外显子组测序研究已显着识别出 100 多个基因与 ASD 风险相关。 ASD 风险基因的功能特征可以提供洞察力自闭症谱系障碍发病机制。我们和其他人最近发现了新的功能丧失变异转录因子 RFX3 是 ASD 相对常见的单基因病因，这意味着 RFX3 在人类神经发育中发挥重要作用。我们发现证据表明 RFX3 可能是 II/III 层神经元发育和功能的关键转录调节因子：表达在皮质 II/III 层兴奋性神经元中显着富集，并且 RFX3 结合基序特别富集于人类胎儿生发区的可接近染色质区域和 II/III 层兴奋性神经元。在这项拟议的研究中，我将提出以下假设： RFX3 调节 II/III 层兴奋性神经元和其他影响神经元形成和功能的 ASD 风险基因的表达。在目标 1 中，我将通过分析人类皮质神经元中 RFX3 调节的基因和通路 RFX3 的全基因组结合位点以及 RFX3 丢失引起的转录变化 RFX3 单倍体中人类 iPSC 衍生神经元的占有率不足。在目标 2 中，我将评估 RFX3单倍体不足对人类皮质神经元形成和突触功能的影响 iPSC 衍生的前脑类器官。我将使用单细胞 RNA 测序来识别细胞的变化类型组成并推断 RFX3 缺陷类器官发育轨迹的变化，和多电极阵列评估 RFX3 缺陷类器官的突触可塑性平衡与同基因对照相比。总而言之，该提案将产生对人类神经元中 RFX3 调节的转录程序，以及 RFX3 单倍体不足如何破坏神经元的发育和功能。这将有助于加深对自闭症谱系障碍 (ASD) 的了解神经生物学，以及 ASD 新型靶向疗法的开发。