Deciphering the roles of COMPASS-related methyltransferases in intellectual and developmental disability

解读 COMPASS 相关甲基转移酶在智力和发育障碍中的作用

• 批准号: 10505308
• 负责人: Ryan Dhindsa
• 金额: $ 6.49万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2023-07-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10505308
• 关键词: Affect; Area; Automobile Driving; Binding; Binding Sites; Brain; CRISPR interference; CRISPR/Cas technology; Calcium; Candidate Disease Gene; Cell Culture Techniques; Chromatin; Collaborations; Communication; Complex; Data; Disabled Persons; Disease; Environment; Etiology; Family; Fellowship; Functional disorder; Future; Gene Expression; Gene Expression Profiling; Genes; Genetic Heterogeneity; Genetic Transcription; Genomics; Goals; Grant; Health Expenditures; Human; Image; Induced pluripotent stem cell derived neurons; Institutes; Intellectual functioning disability; Lead; Location; MLL gene; Manuscripts; Maps; Medicine; Methods; Methylation; Methyltransferase; Molecular; Morphology; Mutation; Neurons; Oral; Outcome; Pathogenicity; Pathway interactions; Patients; Pattern; Preparation; Protein Methyltransferases; Proteins; Research; Research Personnel; Research Technics; Role; Subgroup; Supervision; Syndrome; System; Testing; Therapeutic; Tissue-Specific Gene Expression; Training; Transcriptional Regulation; Work; Writing; base; career; chromatin remodeling; college; design; epigenomics; experimental study; gene discovery; genetic architecture; histone methylation; histone modification; individualized medicine; induced pluripotent stem cell; insight; knock-down; methylation pattern; multi-electrode arrays; multiple omics; neurophysiology; novel; relating to nervous system; single-cell RNA sequencing; stem cells; student mentoring; targeted treatment; therapeutic target; transcriptomics

Project Summary Roughly 1,000 genes have been associated with intellectual and developmental disability (IDD), but targeted treatment options are lacking. Despite this genetic heterogeneity, many implicated genes have similar functions, suggesting they converge on shared mechanisms. There is a critical need to identify potential convergent mechanisms to develop therapeutics for each subclass of genes associated with IDD. This proposal focuses on six related chromatin regulators to determine whether IDD-causing mutations in these genes affect similar molecular pathways. These six SET methyltransferases (SETD1A, SETD1B, KMT2A, KMT2B, KMT2C, and KMT2D) each form a separate COMPASS complex that methylates H3K4. Haploinsufficiency of each gene causes a separate neurodevelopmental syndrome with intellectual disability as a feature. My central hypothesis is that mutations in these COMPASS-related methyltransferases cause intellectual and developmental disability through dysregulating a shared set of genes due to changes in H3K4 methylation patterns. The alternative hypothesis is that IDD results from broad gene expression dysregulation of non-overlapping gene sets. The proposed aims will test both hypotheses and the data I will generate will provide new insight into mechanisms driving IDD irrespective of the outcome. I will use a robust CRISPR interference-based platform in human induced pluripotent stem cells (iPSCs) to knockdown and study these six COMPASS-related genes. In Aim 1, I will elucidate dysregulated molecular pathways due to haploinsufficiency of COMPASS methyltransferases via transcriptomic and epigenomic profiling. These data will allow me to test the hypothesis that loss of these genes leads to dysregulation of similar gene sets and determine whether these similarities can be explained by shared genomic binding sites. In Aim 2, I will functionally characterize morphological and neurophysiological effects of COMPASS methyltransferase haploinsufficiency on human iPSC-derived neurons. These experiments will reveal either common or divergent signatures at the cellular and network levels and will provide insight into the pathophysiology of intellectual disability. This proposal is significant because it will establish a framework to elucidate convergent mechanisms in IDD that could be scaled to a broader set of genes in future studies. My fellowship proposal also outlines a rigorous training plan focused on five specific areas to support my long-term career goal of becoming an independent investigator: (1) identifying interesting research questions and designing rigorous experiments; (2) effective scientific communication in oral presentations, manuscript writing, and grant writing; (3) novel research techniques, including CRISPR methods, stem cell culture, calcium imaging, and chromatin profiling; (4) mentoring students and collaborating with colleagues; and (5) preparation for an independent career. Through direct supervision from Dr. Huda Zoghbi and Dr. Zhandong Liu, I will be immersed in an extremely collaborative and supportive training environment at Baylor College of Medicine.

项目摘要 大约有1,000个基因与智力和发育障碍（IDD）有关，但针对性 缺乏治疗选择。尽管这种遗传异质性，但许多牵连的基因具有相似的功能， 表明它们会融合共同的机制。肯定需要确定潜在的收敛性 为与IDD相关的每个基因子类开发治疗剂的机制。该提议重点 六个相关的染色质调节剂，以确定这些基因中引起IDD的突变是否影响相似 分子途径。这六套甲基转移酶（setD1a，setd1b，kmt2a，kmt2b，kmt2c，and kmt2c和 kmt2d）每个形成一个甲基化H3K4的单独的指南针复合物。每个基因的单倍不足 引起单独的神经发育综合征，其智力障碍作为特征。我的中心假设 是这些与指南针相关的甲基转移酶的突变会导致智力和发育障碍 由于H3K4甲基化模式的变化，通过失调的一组共享基因。替代方案 假设是IDD是由非重叠基因集的广泛基因表达失调引起的。这 拟议的目标将同时检验假设，我将生成的数据将提供有关机制的新见解。 驾驶IDD不管结果如何。 我将在人类诱导的多能干细胞（IPSC）中使用强大的CRISPR干扰平台 敲低并研究了这六个与指南针相关的基因。在AIM 1中，我将阐明分子失调 通过转录组和表观基因组学的指南针甲基转移酶的单倍不足导致的途径 分析。这些数据将使我能够检验以下假设：这些基因的丧失会导致类似的失调 基因集并确定这些相似性是否可以通过共享的基因组结合位点来解释。在AIM 2中， 我将在功能上表征指南针甲基转移酶的形态学和神经生理效应 对人IPSC衍生的神经元的单倍不足。这些实验将揭示常见或发散 在细胞和网络级别的签名，将提供有关智力的病理生理学的见解 残疾。该提议很重要，因为它将建立一个阐明收敛机制的框架 在IDD中，可以在未来的研究中缩放到更广泛的基因。我的奖学金提议还概述了 严格的培训计划着重于五个特定领域，以支持我成为一个成为一个长期职业目标 独立研究者：（1）确定有趣的研究问题并设计严格的实验； （2） 在口头演示，手稿写作和赠款写作中有效的科学沟通； （3）新研究 技术，包括CRISPR方法，干细胞培养，钙成像和染色质分析； （4） 指导学生并与同事合作； （5）为独立职业做准备。通过 Huda Zoghbi博士和Zhandong Liu博士的直接监督，我将沉浸在一个非常合作的中 和贝勒医学院的支持性培训环境。