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 Sotos综合征，其特征是童年过度生长，全球发育迟缓，智力残疾和行为缺陷是由核受体结合SET结构域蛋白1（NSD1）的单倍不足驱动的编码组蛋白H3赖氨酸36（H3K36）甲基转移酶。 NSD1专门催化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对于维持通过募集DNA甲基转移酶3A（DNMT3A）的DNAME，NSD1损耗导致H3K36ME2减少拮抗剂标记组蛋白H3赖氨酸27三甲基化（H3K27me3）的增益，NSD1可能通过促进RNAPII向伸长能力的状态过渡来调节基因表达。细胞表型研究进一步确定了H3K36ME2在细胞塑性和癌症中的重要作用转移。这些机制与Sotos综合征患者的神经发育和功能障碍如何相关，但是，尚待阐明。实际上，NSD1突变和H3K36ME失调的影响是据报道是高度依赖的，导致对基本参数（例如单元）的相反影响跨不同细胞类型的增殖，对人脑中的NSD1功能和功能障碍鲜为人知与SOTOS综合征相关的细胞类型。我们的实验室最近确定了特定而显着的减少与多倍体对照相比我们一直在研究H3K36ME2 / H3K27ME3染色质结合之间的关系，唐氏综合症中的dname和转录本表达。这些数据使我们考虑了有趣的可能性唐氏综合症和索托斯综合征可以共享常见的分子扰动常见的神经发育表型。但是，缺乏研究NSD1功能的工具和数据集在与Sotos综合征相关的环境中（即人类神经发育），已经出现了重大的障碍进步。因此，在AIM I中，我们建议生成一组新型的NSD1的等源性人IPSC模型单倍不足。 NSD1上H3K36ME2 / H3K27ME3染色质结合模式的失调然后，将通过剪切和标记数据集评估人神经元中的单倍不足。严格的该提案的执行将为研究NSD1创造新的灵活和可持续资源单倍不足并测试NSD1单倍不足在生理上相关的分子影响人脑细胞类型。该建议适合R03机制，因为结果将发展出新颖基于人类细胞的模型并生成离散的试验数据，以支持未来的R01提交。