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.

项目概要/摘要 NIH K08 提案描述了癫痫遗传学研究的五年职业发展计划。 通过这个项目，Khoshkhoo 博士将接受人类遗传学、实验和分析方面的培训 下一代测序和单细胞基因组学方面的新技能将补充 Dr. Khoshkhoo 之前的研究和临床培训，使他能够理想地过渡到独立 研究人员研究癫痫遗传变异的功能和分子机制。 布莱根妇女医院 (BWH)、波士顿儿童医院提供的机构资源 医院 (BCH) 和哈佛医学院 (HMS) 是世界一流的，它们提供了理想的环境 促进年轻医师科学家的职业发展。Khoshkhoo 博士是该提案的导师。 Christopher Walsh（HMS 神经病学教授和 BCH HHMI 研究员）是遗传学领域的领军人物 沃尔什博士在指导其他学员进行人类神经系统疾病和基因组学方面有着长期的记录。 此外，Khoshkhoo 博士还组建了一批合作者。 具有互补的专业知识，以及在指导医生方面具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性合子后（体细胞）的作用 颞叶癫痫 (TLE) 的突变（变异），并表征细胞类型特异性和 这些变异导致癫痫发展的转录机制。 Khoshkhoo 提供的试点数据表明，散发性 TLE 病例的一个子集可能含有致病性躯体细胞 变异，这支持了他的中心假设，即遗传和转录失调是由 体细胞变异在 TLE 发病机制中发挥着关键作用。该提案将系统地检查外科 TLE。 切除这些体细胞变异的存在并研究它们的细胞和转录 机制。 为了实现这些目标，结合了超深度基因组测序、平行 RNA 和 DNA 深度测序（PRDD-seq）和单核RNA测序（snRNA-seq）后的分析将 采用这些最先进的策略旨在：（1）在手术中识别致病性体细胞变异。 从 TLE 患者和死后神经典型个体切除海马组织；(2) 确定 具有已知致病性体细胞变异的 TLE 病例中突变细胞的细胞类型；以及 (3) 检查 这些变异在 TLE 中引起下游基因表达变化，这些研究不仅会有所帮助。 建立体细胞变异作为 TLE 发病机制的新机制，同时也研究它们的细胞和 总体而言，该提案引入了一个新的概念和实验框架。 研究 TLE 的结果可能具有直接的诊断和治疗意义。