Transcriptional regulation, neuronal development, and function of the mushroom body in a Drosophila model of intellectual disability

智力障碍果蝇模型中的转录调节、神经元发育和蘑菇体的功能

• 批准号: 10056231
• 负责人: Hayden Alexander Moses Hatch
• 金额: $ 5.1万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-10-05 至 2022-10-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10056231
• 关键词: Affect; Aggressive behavior; Alleles; Anxiety; Axon; Behavioral; Binding; Biological Assay; Brain; Caregivers; Cell Nucleus; Cells; Cellular Morphology; ChIP-seq; Cognition; Cognitive; Consumption; DNA Binding; DNA Binding Domain; Data; Data Set; Defect; Development; Diagnosis; Disease; Drosophila genus; Epilepsy; Family; Functional disorder; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Transcription; Goals; Growth; Image; Impairment; Inherited; Intellectual functioning disability; Knockout Mice; Lead; Learning; Link; Manuscripts; Mediating; Memory; Memory impairment; Mentorship; Missense Mutation; Modeling; Molecular; Morphogenesis; Morphology; Mosaicism; Mus; Mushroom Bodies; Mutation; Nature; Neuroglia; Neurons; Neuropil; Neurosciences; Orthologous Gene; Output; Pathogenesis; Pathway interactions; Patients; Pattern; Physicians; Population; Process; Proteins; RNA Interference; Reporting; Research; Research Personnel; Resolution; Retrieval; Scientist; Seizures; Social Behavior; Stereotyping; Structural defect; Structure; Syndrome; Techniques; Testing; Therapeutic; Time; Training; Transcription Regulatory Protein; Transcriptional Regulation; Vertebral column; Work; X-linked intellectual disability; base; behavioral phenotyping; cell type; cognitive function; differential expression; early childhood; effective therapy; experience; fly; genetic profiling; genome-wide; histone demethylase; in vivo; innovation; knock-down; long short term memory; mouse model; mutant; neuron development; neuronal guidance; novel; programs; relating to nervous system; skills; social; therapeutic development; therapeutically effective; tool; transcriptome; transcriptome sequencing; treatment strategy

ABSTRACT Intellectual disability (ID) disorders affect 2% of the population and are characterized by an IQ score lower than 70 with deficits in adaptive functioning. Mutations in over 400 genes contribute to the pathogenesis of ID disorders, with patients presenting with learning and memory impairments and often syndromic features such as epilepsy, anxiety, short stature, and aggressive tendencies. Our research focusses on the KDM5 family of transcriptional regulators, mutations in which account for 1-3% of inherited ID ranging from mild to severe. The molecular mechanisms by which KDM5 proteins impact neuronal function remain largely unknown, leaving patients without effective treatment strategies. Thus, the overarching goal of this project will be to understand how mutations in KDM5 contribute to neuronal and transcriptional outputs that influence cognition. Here, we will utilize the genetically tractable Drosophila, which encodes a single ortholog of kdm5, to investigate neuronal morphology, transcriptional outputs, and behavioral phenotypes of flies bearing patient-derived KDM5 missense mutations. We have generated a set of ten fly strains, each of which harbors a conserved mutation in Drosophila kdm5 that is analogous to an ID-associated allele. Preliminary data have demonstrated that RNAi-mediated knockdown of kdm5 results in profound guidance and growth defects of the mushroom body (MB), a paired neuropil-rich structure required for the acquisition, consolidation, and retrieval of long- and short-term memory. Significantly, similar morphological MB defects are observed in flies bearing a mutation analogous to an ID- causing missense mutation in an A/T rich interacting domain (ARID) previously implicated in KDM5 DNA binding. Based on these and other data, our central hypothesis is that KDM5 is essential for MB development, and that conserved ID-associated missense mutations that disrupt KDM5’s transcriptional activity lead to the misexpression of genes required for MB morphology and cognitive function. This hypothesis will be tested in three specific aims. The first will be to quantify the extent of MB defects in all kdm5 mutant strains harboring ID- associated missense mutations at both gross and single-cell resolution. The second aim will define the gene expression defects within MB neurons of kdm5 knockdown and ID mutant strains using combined genome-wide transcriptome (RNA-seq) and binding (ChIP-seq) assays. Our third aim will quantify analogous cognitive and behavioral phenotypes that are classically associated with ID. This work will thus be the first to utilize conserved ID-causing KDM5 missense alleles to link ID-associated behavioral phenotypes with neuronal and transcriptional regulatory programs, opening new avenues for the development of therapeutic strategies. Under the mentorship and guidance of Drs. Julie Secombe and Nicholas Baker, I will be able to accomplish these goals while acquiring new skills in developmental neuroscience and related fields. Additionally, I will gain valuable experience presenting, networking, and preparing manuscripts, skills that are essential as I train to become an independent investigator and physician-scientist in the neurosciences.

摘要 智力残疾(ID)障碍影响2%的人口，其特点是智商低于 70人在适应功能方面存在缺陷。400多个基因突变与ID的发病机制有关 精神障碍，患者表现为学习和记忆障碍，通常是症状特征，如 癫痫、焦虑、矮小和攻击性倾向。我们的研究重点是KDM5家族 转录调节因子，突变占遗传性ID的1-3%，从轻微到严重不等。这个 KDM5蛋白影响神经元功能的分子机制在很大程度上仍不清楚， 没有有效治疗策略的患者。因此，该项目的首要目标将是了解 KDM5的突变如何影响影响认知的神经元和转录输出。在这里，我们将 利用遗传上易驯化的果蝇，它编码一个kdm5的同源基因，来研究神经元 携带患者来源KDM5错义的果蝇的形态、转录输出和行为表型 突变。我们已经产生了一组10个果蝇品系，每个品系都含有果蝇的保守突变 Kdm5，类似于ID相关的等位基因。初步数据表明，RNAi介导的 敲除kdm5导致蘑菇体(MB)的严重引导和生长缺陷，配对的 获取、巩固和提取长期和短期记忆所需的富含神经纤维层的结构。 值得注意的是，在携带与ID类似的突变的果蝇中也观察到了类似的MB形态缺陷。 导致富含A/T相互作用结构域(ARID)的错义突变，该结构域以前与KDM5 DNA结合有关。 基于这些和其他数据，我们的中心假设是，KDM5对于甲基溴的开发是必不可少的，而且 保守的ID相关错义突变破坏KDM5‘S转录活性导致 MB形态和认知功能所需的基因表达错误。这一假设将在#年进行检验。 三个具体目标。第一个是量化所有携带ID-1的kdm5突变株中MB缺陷的程度。 相关的大体和单细胞分辨率的错义突变。第二个目标是定义基因 联合全基因组检测kdm5基因敲除株和ID突变株MB神经元中的表达缺陷 转录组(RNA-seq)和结合(芯片-seq)分析。我们的第三个目标将量化类似的认知和 传统上与ID相关的行为表型。因此，这项工作将是第一个利用保守性的 导致ID的KDM5错义等位基因将ID相关的行为表型与神经元和转录联系起来 监管计划，为治疗策略的发展开辟了新的途径。在导师的指导下 在Julie Secombe博士和Nicholas Baker博士的指导下，我将能够在获得 发展神经科学及相关领域的新技能。此外，我还将获得宝贵的经验 展示、建立关系网和准备手稿，这些技能在我成为独立人士的培训过程中至关重要 神经科学领域的研究员和内科科学家。