Genetic and Cellular Mechanisms of Temporal Lobe Epilepsy

颞叶癫痫的遗传和细胞机制

• 批准号: 10661839
• 负责人: Sattar Khoshkhoo
• 金额: $ 24.08万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661839
• 关键词: Address; Advisory Committees; Affect; Autopsy; BRAF gene; Biomedical Research; Blood Vessels; Boston; Brain; Cell physiology; Cells; Chronic; Complement; Conceptions; Cortical Dysplasia; DNA; DNA Sequence Alteration; DNA analysis; Data; Development; Diagnostic; Disease; Environment; Epilepsy; Epileptogenesis; Event; Excision; FRAP1 gene; Fostering; Freezing; Ganglioglioma; Gene Expression; Genes; Genetic; Genetic Research; Genetic Transcription; Genomics; Goals; Hippocampus; Hospitals; Human; Human Genetics; In Situ; Individual; Institution; Lesion; Link; MAP Kinase Gene; MAPK Signaling Pathway Pathway; Magnetic Resonance Imaging; Mediating; Medical; Mentors; Molecular; Morbidity - disease rate; Mosaicism; Mutation; N-Methyl-D-Aspartate Receptors; Neurologic; Neurology; Neurons; Operative Surgical Procedures; Partial Epilepsies; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pediatric Hospitals; Persons; Physicians; Play; Positioning Attribute; Program Development; Publishing; RNA; RNA analysis; Ras/Raf; Refractory; Reporting; Research; Research Personnel; Research Training; Resected; Resistance; Resources; Rodent; Role; Scientist; Sclerosis; Seizures; Signal Transduction; Somatic Mutation; Specificity; Techniques; Temporal Lobe; Temporal Lobe Epilepsy; Tissues; Training; Trauma; United States National Institutes of Health; Variant; Woman; brain tissue; career; career development; cell type; clinical training; comparative; deep sequencing; diagnostic strategy; experience; frontal lobe; gain of function; gene panel; genetic variant; medical schools; mind control; mouse model; mutant; nervous system disorder; neuropsychiatric disorder; neurotrophic factor; next generation sequencing; novel; overexpression; professor; programs; single nucleus RNA-sequencing; skills; transcriptome; transcriptome sequencing; tumor

PROJECT SUMMARY / ABSTRACT This NIH K08 proposal, describes a five-year career development program in epilepsy genetics research. Through this program, Dr. Khoshkhoo will receive training in human genetics, the experimental and analytic aspects of next generation sequencing, and single cell genomics. This new skillset will complement Dr. Khoshkhoo’s prior research and clinical training, and ideally position him to transition to an independent investigator position studying the functional and molecular mechanisms of genetic variants in epilepsy. The institutional resources available through Brigham and Women’s Hospital (BWH), Boston Children’s Hospital (BCH), and Harvard Medical School (HMS) are world class and they provide the ideal environment to foster the career developmental of young physician-scientists. Dr. Khoshkhoo’s mentor for this proposal, Dr. Christopher Walsh (a Professor of Neurology at HMS and HHMI Investigator at BCH), is a leader in genetics and genomics of human neurologic diseases. Dr. Walsh has a long track record for mentoring other trainees to successful careers in biomedical research. In addition, Dr. Khoshkhoo has assembled a group of collaborators with complementary expertise, and an Advisory Committee with extensive experience in mentoring physician- scientists to develop independent research programs. The primary scientific objective of this proposal is to identify the role of pathogenic post-zygotic (somatic) mutations (variants) in temporal lobe epilepsy (TLE), and to characterize the cell-type specific and transcriptional mechanisms through which these variants contribute to the development of epilepsy. Dr. Khoshkhoo provides pilot data indicating that a subset of sporadic TLE cases harbor likely pathogenic somatic variants, which supports his central hypothesis that genetic and transcriptional dysregulation caused by somatic variants plays a key role in TLE pathogenesis. This proposal will systemically examine surgical TLE resections for the presence of these somatic variants and investigate their cellular and transcriptional mechanisms. To achieve these objectives, a combination of ultra-deep gene panel sequencing, Parallel RNA and DNA analysis after Deep sequencing (PRDD-seq), and single nucleus RNA sequencing (snRNA-seq) will be employed. These state of the art strategies will aim to: (1) identify pathogenic somatic variants in surgically resected hippocampal tissue from TLE patients and post-mortem neurotypical individuals; (2) determine the cell-type(s) of mutant cells in TLE cases with known pathogenic somatic variants; and (3) examine the downstream gene expression changes caused by these variants in TLE. These studies will not only help establish somatic variants as a novel mechanism for TLE pathogenesis, but also investigate their cellular and molecular mechanisms in situ. Overall, this proposal introduces a new conceptual and experimental framework for studying TLE and the findings may have immediate diagnostic and treatment implications.

项目摘要 /摘要 该NIH K08提案描述了癫痫遗传学研究的五年职业发展计划。 通过该计划，Khoshkhoo博士将接受人类遗传学的培训，实验和分析 下一代测序和单细胞基因组学的各个方面。这个新技能将完成博士。 Khoshkhoo的先前研究和临床培训，理想情况下，他将其定位为独立 研究者的位置研究了癫痫中遗传变异的功能和分子机制。 波士顿儿童的Brigham and妇女医院（BWH）可获得的机构资源 医院（BCH）和哈佛医学院（HMS）是世界一流，它们为 促进年轻医师科学家的职业发展。 Khoshkhoo博士对该建议的精神，博士 克里斯托弗·沃尔什（HMS的神经病学教授和BCH的HHMI研究者）是遗传学的领导者 和人类神经疾病的基因组学。沃尔什博士对其他学员的心理记录很长 生物医学研究的成功职业。此外，Khoshkhoo博士已经组建了一群合作者 具有完善的专业知识和一个在心理医师方面拥有丰富经验的咨询委员会 - 科学家制定独立的研究计划。 该提案的主要科学目标是确定致病后杂志（体细胞）的作用 临时叶癫痫（TLE）中的突变（变体），并表征细胞类型的特异性和 这些变体有助于癫痫发展的转录机制。博士 Khoshkhoo提供了试验数据，表明一部分零星的案例可能具有致病性的体细胞 变体支持他的中心假设，即遗传和转录失调由 体细胞变体在TLE发病机理中起关键作用。该建议将系统地检查手术 这些体细胞变异的分辨率并研究了它们的细胞和转录 机制。 为了实现这些目标，超深基因面板测序，平行RNA和DNA的结合 深度测序后（PRDD-SEQ）和单核RNA测序（SNRNA-SEQ）分析将是 雇用。这些艺术策略的目的是：（1）通过外科手术确定致病的体细胞变异 从TLE患者和验尸神经型个体中切除的海马组织； （2）确定 突变细胞的细胞类型在具有已知病原性体细胞变异的TLE病例中； （3）检查 这些变体在TLE中引起的下游基因表达变化。这些研究不仅会有所帮助 建立躯体变体作为TLE发病机理的一种新型机制，但也研究其细胞和 原位分子机制。总体而言，该建议引入了一个新的概念和实验框架 用于研究TLE和发现可能具有直接的诊断和治疗意义。