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Establishing foundational tools and datasets for investigation of NSD1 gene function in neural development

建立用于研究神经发育中 NSD1 基因功能的基础工具和数据集

基本信息

批准号：
10711291
负责人：
Lindy Elise Barrett
金额：
$ 15.8万
依托单位：
BROAD INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-15 至 2024-08-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10711291
关键词：
Acceleration Behavioral Binding CRISPR/Cas technology Cancer cell line Cell Line Cell Proliferation Cells Childhood Chromatin DNA Maintenance DNA Methylation DNA Modification Methylases Data Data Set Developmental Delay Disorders Disease Down Syndrome Euchromatin Functional disorder Future Gene Expression Genes Genetic Transcription Growth Growth Disorders Histone H3 Human Induced pluripotent stem cell derived neurons Intellectual functioning disability Intercistronic Region Investigation Laboratories Lysine Malignant Neoplasms Methylation Methyltransferase Modeling Molecular Mus Muscle hypotonia Mutation Neoplasm Metastasis Neurobiology Neurodevelopmental Disorder Neuronal Dysfunction Neurons Nuclear Receptors Patients Pattern Phenotype Physiological Play Preparation Reporting Research Resources Role SET Domain Sotos syndrome Tertiary Protein Structure Testing Time Transcript autosome brain cell cell type flexibility gene function genetic manipulation histone methyltransferase induced pluripotent stem cell insight neurodevelopment novel pharmacologic receptor binding recruit stem cell model sustainable resource tool

项目摘要

SUMMARY Sotos syndrome, characterized by childhood overgrowth, global developmental delay, intellectual disability and behavioral deficits is driven by haploinsufficiency of Nuclear Receptor Binding SET Domain Protein 1 (NSD1) which encodes a histone H3 lysine 36 (H3K36) methyltransferase. NSD1 specifically catalyzes H3K36 dimethylation (H3K36me2), which has important roles in DNA methylation (DNAme) and transcription. Previous studies using mouse or cancer cell lines have shown that H3K36me2 is essential for maintenance of DNAme via recruitment of DNA Methyltransferase 3A (DNMT3A), that NSD1 loss leads to reduced H3K36me2 and gain of the antagonistic mark histone H3 lysine 27 trimethylation (H3K27me3), and that NSD1 may regulate gene expression by facilitating the transition of RNAPII to an elongation-competent state. Cellular phenotyping studies have further identified important roles for H3K36me2 in cellular plasticity and cancer metastasis. How these mechanisms relate to neural development and dysfunction in Sotos syndrome patients, however, remains to be elucidated. Indeed, the effects of NSD1 mutation and H3K36me dysregulation are reported to be highly context dependent, resulting in opposing impacts on basic parameters such as cell proliferation across different cell types, with little known about NSD1 function and dysfunction in human brain cell types relevant for Sotos syndrome. Our laboratory recently identified a specific and significant decrease in H3K36me2 across Down syndrome (driven by trisomy 21) patient cell lines compared to euploid controls which we have been investigating by probing the relationship between H3K36me2 / H3K27me3 chromatin binding, DNAme and transcript expression in Down syndrome. These data led us to consider the intriguing possibility that Down syndrome and Sotos syndrome could share common molecular perturbations contributing to common neurodevelopmental phenotypes. However, the dearth of tools and datasets to study NSD1 function in a context relevant for Sotos syndrome (i.e., human neural development), has presented a significant barrier to progress. In Aim I, we therefore propose to generate a set of novel isogenic human iPSC models of NSD1 haploinsufficiency. Dysregulation of H3K36me2 / H3K27me3 chromatin binding patterns upon NSD1 haploinsufficiency in human neurons will then be assessed through pilot CUT&Tag datasets. Rigorous execution of this proposal will create new flexible and sustainable resources for studying NSD1 haploinsufficiency and test the molecular impacts of NSD1 haploinsufficiency in a physiologically relevant human brain cell type. This proposal is appropriate for the R03 mechanism, as the results will develop novel human cell-based models and generate discrete pilot data to support a future R01 submission.

概括索托斯综合征，其特征是儿童期过度生长、整体发育迟缓、智力障碍和行为缺陷是由核受体结合 SET 结构域蛋白 1 (NSD1) 的单倍体不足引起的它编码组蛋白 H3 赖氨酸 36 (H3K36) 甲基转移酶。 NSD1 特异性催化 H3K36 二甲基化 (H3K36me2)，在 DNA 甲基化 (DNAme) 和转录中具有重要作用。先前使用小鼠或癌细胞系的研究表明，H3K36me2 对于维持 DNAme 通过招募 DNA 甲基转移酶 3A (DNMT3A)，NSD1 丢失导致 H3K36me2 减少和拮抗标记组蛋白 H3 赖氨酸 27 三甲基化 (H3K27me3) 的获得，并且 NSD1 可能通过促进 RNAPII 向延伸状态的转变来调节基因表达。蜂窝网络表型研究进一步确定了 H3K36me2 在细胞可塑性和癌症中的重要作用转移。这些机制与索托斯综合征患者的神经发育和功能障碍有何关系，然而，仍有待阐明。事实上，NSD1 突变和 H3K36me 失调的影响是据报道高度依赖于环境，导致对细胞等基本参数产生相反的影响跨不同细胞类型的增殖，但对人脑中 NSD1 的功能和功能障碍知之甚少与索托斯综合征相关的细胞类型。我们的实验室最近发现了特定且显着的下降唐氏综合症（由 21 三体驱动）患者细胞系中的 H3K36me2 与整倍体对照相比，我们一直在通过探究 H3K36me2 / H3K27me3 染色质结合之间的关系进行研究，唐氏综合症中的 DNAme 和转录本表达。这些数据让我们考虑了一个有趣的可能性唐氏综合症和索托斯综合症可能有共同的分子扰动，导致常见的神经发育表型。然而，缺乏研究 NSD1 功能的工具和数据集在与索托斯综合症（即人类神经发育）相关的背景下，已经提出了一个重大障碍进步。因此，在目标 I 中，我们建议生成一组新的 NSD1 等基因人类 iPSC 模型单倍体不足。 NSD1 上 H3K36me2 / H3K27me3 染色质结合模式的失调然后，将通过试点 CUT&Tag 数据集评估人类神经元的单倍体不足。严格的该提案的执行将为研究 NSD1 创建新的灵活且可持续的资源单倍体不足并测试 NSD1 单倍体不足在生理相关的分子影响人类脑细胞类型。该提议适用于 R03 机制，因为结果将发展出新颖的基于人类细胞的模型并生成离散的试点数据以支持未来的 R01 提交。