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：
10535366
负责人：
Jenny Lai
金额：
$ 3.9万
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/10535366
关键词：
Address Affect Attentional deficit 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 Hyperactivity 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 Research Research Proposals Risk Role Signal Transduction Social Interaction Statistical Data Interpretation Synapses Synaptic Transmission Synaptic plasticity Testing Time Tissues Training Variant autism spectrum disorder cell type ciliopathy cohort conditional knockout de novo mutation differential expression disorder risk excitatory neuron exome sequencing fetal genome sequencing genome-wide 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 neurobiological mechanism neurodevelopment neuron development new therapeutic target next generation novel postnatal human programs promoter relating to nervous system 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发病机理上的神经生物学机制在很大程度上未知。大规模 ASD个体的外显子组测序研究已显着鉴定出100多种基因与ASD风险相关。 ASD风险基因的功能表征可以为您提供见识 ASD发病机理。我们和其他人最近确定了从头丧失的功能丧失变体转录因子RFX3是ASD的相对常见的单基因原因，这意味着 RFX3在人神经发育中的重要作用。我们发现RFX3可能成为II/III层神经元发展和功能的关键转录调节剂：它表达在皮质II/III层兴奋性神经元和RFX3结合中显着富集基序特别丰富了人类胎儿生发区的可访问染色质区域和II/III层兴奋性神经元。在这项拟议的研究中，我将解决以下假设 RFX3调节II/III层兴奋性神经元中的关键神经发育过程和影响神经元形成和功能的其他ASD风险基因的表达。在AIM 1中，我会通过分析人类皮质神经元中RFX3调节的基因和途径 RFX3的全基因组结合位点以及损失RFX3引起的转录变化 RFX3中的占用率是人IPSC衍生的神经元。在AIM 2中，我将评估 RFX3单倍宽度对人类皮质神经元形成和突触功能的影响 IPSC衍生的前脑器官。我将使用单细胞RNA-setering来识别细胞的变化类型组成和推断RFX3缺乏器官中发育轨迹的改变，和多电极阵列，以评估RFX3缺乏类的类器官的突触可塑性平衡与等源对照相比。综上所述，该提议将对由RFX3在人类神经元中调节的转录程序，以及RFX3单倍不足效率破坏神经元的发育和功能。这将使人们对ASD的了解神经生物学以及新型ASD的靶向疗法的发展。