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.

项目概要 大约 1,000 个基因与智力和发育障碍 (IDD) 有关，但有针对性 缺乏治疗选择。尽管存在遗传异质性，但许多相关基因具有相似的功能， 表明它们趋于共享机制。迫切需要识别潜在的收敛性 为与 IDD 相关的每个基因亚类开发治疗方法的机制。该提案的重点是 六个相关的染色质调节因子，以确定这些基因中引起 IDD 的突变是否会影响类似的 分子途径。这六种 SET 甲基转移酶（SETD1A、SETD1B、KMT2A、KMT2B、KMT2C 和 KMT2D) 各自形成一个单独的 COMPASS 复合物，使 H3K4 甲基化。每个基因的单倍体不足 导致以智力障碍为特征的单独的神经发育综合征。我的中心假设 这些 COMPASS 相关甲基转移酶的突变会导致智力和发育障碍 由于 H3K4 甲基化模式的变化而导致一组共享基因失调。另一种选择 假设 IDD 是由非重叠基因组的广泛基因表达失调引起的。这 提出的目标将测试这两个假设，我将生成的数据将为机制提供新的见解 无论结果如何，驾驶 IDD。 我将在人类诱导多能干细胞 (iPSC) 中使用基于 CRISPR 干扰的强大平台 敲除并研究这六个 COMPASS 相关基因。在目标 1 中，我将阐明失调的分子 COMPASS 甲基转移酶单倍体不足导致的转录组和表观基因组通路 分析。这些数据将使我能够检验以下假设：这些基因的丢失会导致类似基因的失调。 基因集并确定这些相似性是否可以通过共享基因组结合位点来解释。在目标 2 中， 我将从功能上表征 COMPASS 甲基转移酶的形态和神经生理学效应 人类 iPSC 衍生神经元的单倍体不足。这些实验将揭示共同点或不同点 细胞和网络层面的签名，并将提供对智力病理生理学的深入了解 残疾。该提案意义重大，因为它将建立一个框架来阐明趋同机制 在 IDD 中，在未来的研究中可以扩展到更广泛的基因组。我的研究金提案还概述了 严格的培训计划侧重于五个特定领域，以支持我成为一名长期职业目标 独立研究者：（1）确定有趣的研究问题并设计严谨的实验； (2) 在口头报告、手稿写作和资助写作中进行有效的科学交流； (3)新颖的研究 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) 为独立职业做好准备。通过 在 Huda Zoghbi 博士和刘占东博士的直接监督下，我将沉浸在高度协作的氛围中 贝勒医学院的支持性培训环境。