MOLECULAR MECHANISMS UNDERLYING INTELLECTUAL DISABILITY CAUSED BY MUTATIONS IN THE CHROMATIN MODIFIER KDM5C

染色质修饰符 KDM5C 突变导致智力障碍的分子机制

• 批准号: 10455680
• 负责人: Julie Secombe
• 金额: $ 20.65万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-23 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10455680
• 关键词: Address; Affect; Alleles; Animal Model; Attenuated; Biological Assay; Biological Models; Brain; CRISPR/Cas technology; Caregivers; Cell Line; Cell model; Cells; Cerebrum; ChIP-seq; Chromatin; Chromatin Remodeling Factor; Clinical; Code; Cognition; Cognitive; Cognitive deficits; Complement; Core Facility; Defect; Development; Developmental Disabilities; Diagnosis; Disease; Disease model; Drosophila genus; Drosophila melanogaster; Epigenetic Process; Etiology; Fragile X Syndrome; Gene Expression; Gene Mutation; Gene Proteins; Genes; Genetic; Genetic Transcription; Genomics; Goals; Histone H3; Homeostasis; Homologous Gene; House mice; Human; In Vitro; Individual; Induced pluripotent stem cell derived neurons; Inherited; Intellectual and Developmental Disabilities Research Centers; Intellectual functioning disability; Interdisciplinary Study; Jordan; Knockout Mice; Knowledge; Lead; Link; Lysine; Massive Parallel Sequencing; Mediating; Methylation; Modeling; Molecular; Morphology; Mus; Mutation; Neurodevelopmental Disorder; Neurologic; Neurologic Deficit; Neurons; Organoids; Pathologic; Patients; Pharmacology; Phenotype; Play; Process; Proteins; Publications; Publishing; Quality of life; Regulation; Research; Research Project Grants; Ribosomal Proteins; Role; Source; Syndrome; System; TNFRSF5 gene; Techniques; Testing; Transcript; Translations; Work; analytical tool; base; behavioral phenotyping; chromatin immunoprecipitation; chromatin modification; clinical database; clinically relevant; cohort; comorbidity; database of Genotypes and Phenotypes; early childhood; fetal; fly; grasp; histone demethylase; improved; in vitro testing; in vivo; induced pluripotent stem cell; innovation; insight; loss of function mutation; mouse model; multiple omics; mutation correction; neuronal circuitry; novel therapeutics; programs; recruit; ribosome profiling; single-cell RNA sequencing; tool; transcriptomics

PROJECT SUMMARY/ABSTRACT – RESEARCH PROJECT Mutations in the gene encoding the transcriptional regulator lysine demethylase 5C (KDM5C) are found in patients with intellectual disability (ID). While the direct link between loss of function mutations in KDM5C and ID is clear, how KDM5C functions to mediate critical neuronal processes, and therefore the consequence of mutations for mechanisms of IDD, remains unknown. The goal of this proposal is to understand the relationship between KDM5C-regulated gene expression programs and the occurrence of ID and additional comorbid features that are observed in patients. We will achieve this by bringing together a multi-disciplinary research team with expertise in complementary analytical tools and model systems to test two hypotheses. Aim 1 tests the hypothesis that the use of human iPSC-derived in vitro cell models of KDM5C-induced ID will result in the identification of clinically relevant gene expression changes and neuronal functional deficits. One key model system we will use is iPSC-derived cerebral organoids, which recapitulate structural and molecular aspects of fetal brain development and are a critical research tool used to define the underlying cause(s) of neurodevelopmental disorders. Indeed, molecular and cellular studies using in vitro organoid systems allow us to carry out studies in a human cell context that would simply not be possible in vivo. iPSCs and organoids will be generated from two sources: (1) Cells from patients with KDM5C-induced ID from a newly recruited cohort of individuals from which we are generating a genotype-phenotype database; (2) CRISPR-Cas9-mediated gene editing to generate a KDM5C null allele and a published ID allele (KDM5CA388P) that lacks histone demethylase activity using existing iPSC lines generated from typically developing controls. We will use this system to combine morphological, functional and multi-OMICS approaches to define the impact of patient- associated mutations in KDM5C. Aim 2 tests the hypothesis that the regulation of translation efficiency in neurons by the fly homolog of KDM5C is conserved in mammalian systems and that this function is important for cognition. Here we take advantage of fly and mouse animal model systems, both as discovery tools and to test hypotheses regarding possible contributors to the cognitive effects of mutations in KDM5C. Because other inherited forms of ID have altered translation and correcting this deficit has shown promise in mouse models of other ID disorders, we will test whether altered translation similarly plays a key role in a mouse model of KDM5C-induced ID. This work is significant because we will define the etiological links between mutations in human KDM5C and ID. The proposed studies are technologically innovative in the use of complementary model systems and state-of-the-art genomics techniques such as single cell transcriptomics (scRNA-seq). It is also conceptually innovative in proposing a role for translation in KDM5C-induced ID.

项目摘要/摘要 - 研究项目 编码转录调节因子赖氨酸脱甲基酶 5C (KDM5C) 的基因突变存在于 智力障碍（ID）患者。虽然 KDM5C 功能丧失突变与 ID 很清楚，KDM5C 如何发挥作用来介导关键的神经元过程，以及因此产生的结果 IDD 机制的突变仍然未知。该提案的目标是了解 KDM5C调控基因表达程序与ID及其他疾病发生的关系 在患者中观察到的共病特征。我们将通过汇集多学科的力量来实现这一目标 研究团队拥有互补分析工具和模型系统的专业知识来检验两个假设。 目标 1 检验以下假设：使用人类 iPSC 衍生的 KDM5C 诱导 ID 体外细胞模型将 导致临床相关基因表达变化和神经元功能缺陷的识别。一 我们将使用的关键模型系统是 iPSC 衍生的大脑类器官，它概括了结构和分子 胎儿大脑发育的各个方面，是用于确定胎儿大脑发育的根本原因的重要研究工具 神经发育障碍。事实上，使用体外类器官系统的分子和细胞研究使我们能够 在人体细胞环境中进行在体内根本不可能实现的研究。 iPSC 和类器官将 由两个来源生成：(1) 来自新招募队列的 KDM5C 诱导 ID 患者的细胞 我们从中生成基因型-表型数据库的个体； (2) CRISPR-Cas9介导 基因编辑生成 KDM5C 无效等位基因和已发表的缺少组蛋白的 ID 等位基因 (KDM5CA388P) 使用从典型开发对照产生的现有 iPSC 系进行去甲基酶活性。我们将使用这个 系统结合形态学、功能和多组学方法来定义患者的影响 KDM5C 的相关突变。目标 2 检验以下假设：翻译效率的调节 KDM5C 的果蝇同源神经元在哺乳动物系统中是保守的，并且该功能很重要 为了认知。在这里，我们利用苍蝇和小鼠动物模型系统，既作为发现工具，又 测试有关 KDM5C 突变认知影响的可能因素的假设。因为其他 ID 的遗传形式改变了翻译，纠正这种缺陷已在小鼠模型中显示出希望。 其他 ID 障碍，我们将测试改变的翻译是否同样在小鼠模型中发挥关键作用 KDM5C 诱导的 ID。 这项工作意义重大，因为我们将定义人类 KDM5C 突变之间的病因学联系 和身份证。拟议的研究在使用互补模型系统方面具有技术创新性 最先进的基因组学技术，例如单细胞转录组学 (scRNA-seq)。概念上也是如此 创新性地提出了翻译在 KDM5C 诱导的 ID 中的作用。