Identification and molecular characterization of somatic mutations in MCD

MCD 体细胞突变的鉴定和分子特征

基本信息

批准号：
10666977
负责人：
Peter B Crino
金额：
$ 72.1万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-30 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10666977
关键词：
AKT Signaling Pathway Address Adult Affect Award Biological Markers Biological Models Brain Candidate Disease Gene Cell Proliferation Cerebral cortex Child Clustered Regularly Interspaced Short Palindromic Repeats Collection Copy Number Polymorphism Cortical Dysplasia Cortical Malformation DNA Sequence Alteration DNA sequencing Data Development Disease Early Diagnosis Early treatment Electroporation Embryo Enrollment Epilepsy Excision FRAP1 gene Fertilization Fetal Development Foundations Funding Future Genes Genetic Genetic Risk Genetic study Germ Cells Goals Grant Guide RNA Human Hyperplasia In Vitro Individual Induced pluripotent stem cell derived neurons Inherited Intellectual functioning disability Intractable Epilepsy Knock-out Lead Medical Microgyria Modeling Molecular Morphology Mus Mutation Neocortex Neurons PIK3CG gene Partial Epilepsies Pathogenicity Pathway interactions Patients Phenotype Process Prognosis Protein Subunits Ras Signaling Pathway Research Resected Risk Role Seizures Series Short Tandem Repeat Signal Transduction Signaling Protein Site Somatic Mutation Specimen Structural defect Tissues Translating Variant Work base brain malformation brain tissue cell motility clinical phenotype de novo mutation disease mechanisms study disease-causing mutation exome exome sequencing gene discovery genetic architecture genome sequencing hemimegalencephaly improved in utero in vitro Model in vivo in vivo Model knockout gene loss of function malformation in cortical development migration mind control neocortical nerve stem cell neuroblastoma cell novel prevent public health relevance screening software development stem cells targeted sequencing whole genome zygote

项目摘要

PROJECT SUMMARY Genetic mutations causing disease may be inherited, newly acquired in parental gametes and present in the zygote, or acquired at some point in development after fertilization. The burden and localization of a post- zygotically acquired mutation depends on when the mutation arises. Malformations of cortical development (MCD) are a group of disorders characterized by a range of morphological and structural abnormalities of the cerebral cortex reflecting errors in embryonic cortical development. MCD are associated with refractory epilepsy as well as intellectual disability and may require the surgical removal of the affected tissue for seizure control. There is increasing recognition that post-zygotically acquired somatic mutations occurring in neuroglial progenitor cells can result in a cortical brain malformation. In the previous funding period of this grant, we made significant progress identifying and molecularly characterizing a series of pathogenic post-zygotically acquired somatic variants in the resected brain tissue of individuals with an epilepsy-associated cortical brain malformation. Most notably we identified and functionally characterized the first gene associated with focal cortical dysplasia type I, SLC35A2, and made significant advancements in the understanding of the somatic genetic landscape across MCD. The overarching objective for the next funding cycle is to continue to identify somatic variants across MCD and to functionally characterize the effects of novel variants associated with different types of MCD on cortical development. In Aim 1, we will continue to collect resected brain tissue specimens from individuals with MCD for high-depth exome or targeted gene sequencing. The goal of Aim 1 is to identify novel genes involved in MCD and to ascertain the subset of exome-negative cases for use in Aim 2. In Aim 2, we will use highly sensitive duplex sequencing to detect very low-level somatic variants and PCR-free whole-genome sequencing to detect somatic short tandem repeat variants and intermediately sized somatic copy number variants in exome-negative MCD cases. The goal of Aim 2 is to determine the contribution of these classes of somatic variants that are routinely missed due to the limitations of standard short-read exome sequencing in the overall somatic genetic risk of MCD. Finally, Aim 3 will evaluate the functional consequences of knocking out newly identified MCD genes harboring somatic loss-of-function variants on neuronal morphology, neuronal migration, and ultimately cerebral cortical development in the developing mouse brain. These studies will: (i) continue our in-depth assessment of the role of somatic mutations across MCD subtypes, (ii) identify novel genes/pathways involved in cortical development, (iii) use complementary in vitro, ex vivo, and in vivo models to understand the role of novel genes implicated cortical development, and (iv) establish biomarkers and platforms that can be used in the future for screening of compounds to possibly prevent or improve the prognosis of MCD.

项目摘要可能遗传引起疾病的基因突变，在父母配子中新获得，并存在于合子，或在受精后的某个开发点获得。职位的负担和本地化 - 合子获得的突变取决于突变何时出现。皮质发育的畸形（MCD）是一组疾病，其特征是一系列形态和结构异常大脑皮层反映了胚胎皮质发育中的误差。 MCD与难治性有关癫痫和智力障碍，可能需要手术去除受影响的组织进行癫痫发作控制。人们越来越认识到，神经乳腺癌发生后获得后获得的体细胞突变祖细胞会导致皮质脑畸形。在这笔赠款的前几个资金期间，我们取得了重大进展，以识别和分子表征一系列致病性的。在具有癫痫相关的皮质大脑的个体切除的脑组织中获得的体细胞变异畸形。最值得注意的是，我们确定并在功能上表征了与焦点相关的第一个基因皮质发育不良I型，SLC35A2，并在理解躯体方面取得了重大进步 MCD的遗传景观。下一个资金周期的总体目标是继续确定 MCD跨MCD的体细胞变体，并在功能上表征与之相关的新型变体的影响在皮质发育方面的不同类型的MCD。在AIM 1中，我们将继续收集切除的脑组织来自具有MCD的个体的高深度外显子或靶向基因测序的标本。目标1的目标是确定与MCD有关的新基因，并确定用于AIM 2的外显阴性病例的子集。在AIM 2中，我们将使用高度敏感的双链测序来检测非常低水平的体细胞变体和无PCR 全基因组测序以检测躯体短串联重复变体和中等大小的体细胞外显性MCD案例中的拷贝数变体。目标2的目标是确定由于标准短读外显子的局限在MCD的总体体细胞遗传风险中进行测序。最后，AIM 3将评估功能后果敲除新鉴定的MCD基因，这些基因具有躯体丧失功能障碍变体的神经元变体形态学，神经元迁移以及最终在发育中的小鼠大脑中的脑皮质发育。这些研究将：（i）继续对MCD跨MCD的体细胞突变作用的深入评估亚型，（ii）确定与皮质发育有关的新基因/途径，（iii）使用互补体外，离体和体内模型了解新型基因的作用涉及皮质发育，并且（iv）建立生物标志物和平台将来可以用于筛选化合物以可能预防或改善MCD的预后。