Role of TRIM71 in neural stem cell biology and congenital hydrocephalus

TRIM71 在神经干细胞生物学和先天性脑积水中的作用

• 批准号: 10313612
• 负责人: Duy Phan
• 金额: $ 3.09万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10313612
• 关键词: Academic skills; Affect; Animals; Biological; Biological Process; Biology; Birth; Brain; Brain Diseases; Caenorhabditis elegans; Cell Culture Techniques; Cell physiology; Cells; Cerebral Ventricles; Cerebrospinal Fluid; Cerebrospinal fluid shunts procedure; Childhood; Clinical; Colony-Forming Units Assay; Communicating Hydrocephalus; Complement; Congenital Hydrocephalus; Coupled; Defect; Development; Developmental Brain Malformation; Diagnostic; Educational workshop; Exhibits; Failure; Fluid Balance; Fluids and Secretions; Functional disorder; Future; Genes; Genetic; Goals; Histology; Homologous Gene; Human; Hydrocephalus; Impairment; In Vitro; Intracranial Pressure; Knowledge; Laboratories; Lead; Learning; Live Birth; Magnetic Resonance Imaging; Measurement; Measures; Medicine; MicroRNAs; Modeling; Molecular; Molecular Genetics; Morbidity - disease rate; Morphogenesis; Mus; Mutant Strains Mice; Mutate; Mutation; Nature; Neonatal; Neuroanatomy; Neurodevelopmental Disorder; Neuroepithelial Cells; Neurosciences; Neurosurgeon; Operative Surgical Procedures; Outcome; Pathogenesis; Pathway interactions; Patients; Phylogeny; Physicians; Physiological; Physiology; Preventive measure; RNA-Binding Proteins; Regulation; Research; Role; Scheme; Scientist; Series; Stem cell pluripotency; Techniques; Testing; Therapeutic; Training; Work; absorption; base; career; cerebrospinal fluid flow; clinical decision-making; de novo mutation; epidermal stem cell; exome sequencing; histological studies; improved; in vitro Assay; insight; mortality; mouse model; multidisciplinary; mutant; mutant mouse model; neocortical; nerve stem cell; nestin protein; neurogenesis; neurogenomics; neuroimaging; novel; novel marker; pre-clinical; precision medicine; prenatal; prognostic; stem cell biology; stem cell differentiation; stem cell population; stem cell proliferation; targeted treatment; transcriptome; ventricular system

PROJECT SUMMARY Congenital hydrocephalus (CH) is the most common developmental malformation of the brain affecting 1/1000 births. CH has been classically attributed to failed cerebrospinal fluid (CSF) homeostasis and therefore treated by surgical CSF diversion, with high morbidity and failure rates. The persistence of post- surgical ventriculomegaly in many patients, often with poor neurodevelopmental outcomes, raises questions about our current paradigms of CH and its treatment. Significant gaps in our understanding of the molecular pathogenesis of CH impede the development of preventive measures, targeted therapies, and improved prognostication. By whole exome sequencing, the Kahle lab has identified heterozygous de novo mutations in the TRIM71/lin-41 as the most common genetic cause of human sporadic CH (Jin et al. 2020, Nature Medicine). TRIM71 encodes an RNA-binding protein first discovered in C. elegans as a heterochronic gene that regulates developmental timing of epidermal stem cells. Despite its remarkable evolutionary conservation across phylogeny and robust expression in prenatal neural stem cells (NSCs), the roles of TRIM71 in mammalian brain development and CH are essentially unknown. In preliminary work, I have shown that mouse lines with the murine homolog of the human TRIM71 CH mutation R608H (Trim71R595H/+) or with conditional NSC-specific Trim71 deletion (Nestin-Trim71fl/fl) both recapitulate the severe neonatal-onset communicating hydrocephalus of human patients harboring TRIM71 mutations. The objective of my proposal is to further characterize these models to increase knowledge of human brain development and CH pathogenesis. I hypothesize TRIM71 mutant ventriculomegaly results not from primary cerebrospinal fluid (CSF) over-accumulation, but rather from impaired neurogenesis related to reduced NSC proliferation and precocious NSC differentiation. Aim 1 will characterize the neuroanatomy and CSF physiology of TRIM71-mutant hydrocephalus using brain magnetic resonance imaging (MRI) and direct measurements of CSF hydrodynamics. Aim 2 will elucidate the cellular mechanisms of TRIM71-mutant hydrocephalus using immunofluorescent studies in Trim71 CH mutant mice complemented by in vitro assays using primary NSC cultures. The demonstration that dysregulated neurogenesis rather than primary CSF over-accumulation underlies some CH cases could have paradigm-changing implications that could lead to improved diagnostic, prognostic, and therapeutic strategies, including the prediction of which CH patients may or may not benefit from neurosurgical CSF shunting. These studies will be conducted as part of our long-term goal to develop precision medicine therapies for CH.

项目摘要 先天性脑积水（CH）是影响大脑的最常见发育畸形 1/1000出生。 CH经典归因于脑脊液失败（CSF）稳态和 因此，通过手术CSF转移治疗，发病率和失败率很高。后期的持久性 许多患者的手术心室肿大，通常神经发育结果差，会引起问题 关于我们当前的CH范式及其治疗。我们对分子的理解差距很大 CH的发病机理阻碍了预防措施的发展，有针对性的疗法并改善了 预测。通过整个外显子组测序，卡尔实验室已经确定了杂合的新作用 TRIM71/LIN-41中的突变是人类零星CH的最常见遗传原因（Jin等。 2020年，自然医学）。 TRIM71编码首先在秀丽隐杆线虫中发现的RNA结合蛋白 调节表皮干细胞发育时机的杂化基因。尽管它出色 在产前神经干细胞（NSC）中跨系统发育和鲁棒表达的进化保守性， TRIM71在哺乳动物脑发育和CH中的作用本质上是未知的。在初步工作中，我 已经表明了与人Trim71 CH突变的鼠同源物R608H的小鼠线 （TRIM71R595H/+）或有条件的NSC特异性TRIM71删除（Nestin-Trim71fl/fl）都概括 携带TRIM71的人类患者的严重新生儿发作的脑积水 突变。我的建议的目的是进一步表征这些模型，以增加对 人脑发育和CH发病机理。 i假设TRIM71突变体室性肿瘤结果不是 来自原发性脑脊液（CSF）过度累积，而是来自与之相关的神经发生受损 NSC增殖和早熟的NSC分化减少。 AIM 1将表征神经解剖学和 使用大脑磁共振成像（MRI）和直接 CSF流体力学的测量。 AIM 2将阐明Trim71突变物的细胞机制 在TRIM71 CH突变小鼠中使用免疫荧光研究的脑积水，并补充体外测定 使用原发性NSC培养物。表明神经发生，而不是主要CSF的证明 过度蓄能的基础一些CH案件可能会改变范式的含义，这可能会导致 改进的诊断，预后和治疗策略，包括CH患者的预测 可能会或可能不会从神经外科CSF分流中受益。这些研究将作为我们的一部分进行 为CH开发精确医学疗法的长期目标。