Deciphering brain mosaicism in drug-resistant epilepsy at cellular resolution

在细胞分辨率下解读耐药性癫痫中的大脑镶嵌现象

基本信息

批准号：
10563335
负责人：
Tracy Ann Bedrosian
金额：
$ 48.6万
依托单位：
RESEARCH INST NATIONWIDE CHILDREN'S HOSP
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：
2023-03-01 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10563335
关键词：
Advanced Development Affect Astrocytes Automobile Driving Brain Cell Lineage Cell Nucleus Cell Separation Cells Child Clinical Cognitive Cortical Malformation Data Detection Development Diagnosis Diagnostic Disease Excision Fluorescence Gene Expression Gene Expression Profile Genetic Genetic Predisposition to Disease Genetic Transcription Genotype Goals Hyperplasia Intractable Epilepsy Knockout Mice Knowledge Mission Molecular Mosaicism Neurons Oligodendroglia Operative Surgical Procedures Patients Phenotype Precision therapeutics Publishing Recurrence Research Resected Resolution Seizures Somatic Mutation Sorting Techniques Testing Therapeutic Tissues Transgenic Mice Translating United States National Institutes of Health Variant behavioral outcome brain cell brain tissue causal variant cell type exome sequencing gene discovery genetic testing genetic variant improved individual patient innovation malformation malformation in cortical development member mosaic mouse model novel novel strategies personalized medicine profiles in patients repository targeted treatment therapeutically effective transcriptomics treatment strategy variant detection

项目摘要

Project Summary PROJECT SUMMARY Although somatic mosaicism is a known cause of malformations of cortical development (MCD), it is still difficult to detect sparse somatic variants in patient tissue and study their molecular and functional effects. The long- term goal of this project is to unravel the cellular and molecular complexity of MCD and translate this knowledge into improved diagnostic and treatment opportunities for patients. The overall objectives in this application are to (i) improve somatic variant detection, (ii) identify transcriptional correlates of somatic variants, and (iii) test functional effects of somatic variants in relevant cell types. The central hypothesis is that somatic variants are enriched in particular cell lineages in MCD and that cell identity is an important determinant of a variant’s molecular and functional effects. The rationale for this project is that understanding somatic variation and its consequences at cell-type resolution will ultimately improve gene discovery for MCD and help advance the field toward targeted therapies for drug-resistant epilepsy associated with MCD. The central hypothesis will be tested by pursuing three specific aims: (1) Improve somatic variant detection in patients with focal cortical malformations, (2) Evaluate cell-type-specific transcriptional effects of somatic mutations, and (3) Functionally validate the cell-type-specific contribution of somatic mutations to disease features. Under the first aim, relevant cell types will be enriched from affected patient tissue and sequenced as a strategy to improve somatic variant detection. For the second aim, integrated single-nuclei genotyping and transcriptomic analyses of patient tissue will be used to evaluate gene expression signatures of somatic mosaicism. The third aim will leverage a novel transgenic mouse model of MCD to drive somatic variation in particular cell types and determine how cell identity contributes to specific components of disease. The research proposed in this application is innovative because it takes a new approach to gene discovery in MCD, leverages leading-edge techniques such as integrated single- nuclei genotyping/transcriptomics, and takes advantage of a unique mouse model generated by the applicant’s lab. The proposed research is significant because it is expected to yield new causal somatic variants for MCD and new knowledge of their molecular and functional effects in appropriate cellular contexts. Ultimately, such knowledge has the potential to provide new opportunities to develop or apply targeted treatments.

项目概要项目概要尽管体细胞嵌合是皮质发育畸形（MCD）的已知原因，但仍然很难检测患者组织中的稀疏体细胞变异并研究其分子和功能影响。该项目的长期目标是阐明 MCD 的细胞和分子复杂性并转化这些知识该应用程序的总体目标是改善患者的诊断和治疗机会。 (i) 改进体细胞变异检测，(ii) 识别体细胞变异的转录相关性，以及 (iii) 测试体细胞变异对相关细胞类型的功能影响的中心假设是体细胞变异是富集于 MCD 中的特定细胞谱系，并且细胞身份是变体的重要决定因素该项目的基本原理是了解体细胞变异及其影响。细胞类型分辨率的结果将最终改善 MCD 的基因发现并有助于推动该领域的发展与 MCD 相关的耐药性癫痫的靶向治疗将得到检验。通过追求三个具体目标：（1）改善局灶性皮质患者的体细胞变异检测畸形，(2) 评估体细胞突变的细胞类型特异性转录效应，以及 (3) 功能验证体细胞突变对疾病特征的细胞类型特异性贡献。将从受影响的患者组织中丰富细胞类型并进行测序，作为改善体细胞变异的策略第二个目标是对患者组织进行综合单核基因分型和转录组分析。将用于评估体细胞嵌合体的基因表达特征。 MCD 转基因小鼠模型可驱动特定细胞类型的体细胞变异并确定细胞身份该申请中提出的研究具有创新性，因为。它采用了 MCD 中基因发现的新方法，利用了诸如集成单细胞核基因分型/转录组学，并利用申请人的实验室生成的独特小鼠模型实验室提出的研究意义重大，因为它有望产生 MCD 的新的因果体细胞变异。最终，它们在适当的细胞环境中的分子和功能效应的新知识。知识有可能为开发或应用针对性治疗提供新的机会。