Diversity Supplement: Single Cell Analyses of Neuroimmune Dysfunctions in the Thalamocortical Circuit in FTLD

多样性补充：FTLD 丘脑皮质回路神经免疫功能障碍的单细胞分析

• 批准号: 10403045
• 负责人: Eric J Huang
• 金额: $ 9.33万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2022-01-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10403045
• 关键词: Age; Aging; Autophagocytosis; Brain; Brain Pathology; Brain region; C9ORF72; Cell Nucleus; Cells; Clinical Research; Coculture Techniques; Cytoplasm; Data; Data Set; Dementia; Development Plans; Disease; Exhibits; Frontotemporal Dementia; Frontotemporal Lobar Degenerations; Functional disorder; GRN gene; Genes; Genetic; Goals; Grant; Homeostasis; Human; Immune; Immunohistochemistry; In Situ Hybridization; In Vitro; Individual; Knockout Mice; Lead; Limbic System; Link; Longevity; Mediating; Mentors; Microglia; Molecular Profiling; Mus; Mutation; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neuroimmune; Neurons; PGRN gene; Pathogenesis; Pathway interactions; Patients; Phenotype; Physiological; Property; Proteins; RNA-Binding Proteins; Reporting; Research; Scientist; Synapses; Testing; Thalamic structure; TimeLine; Ubiquitin; Validation; Western Blotting; Wild Type Mouse; Work; age related; aging brain; astrogliosis; behavioral phenotyping; career; career development; cerebral atrophy; cohort; early onset; glial activation; graduate student; innovation; insight; loss of function; neuroinflammation; neuron loss; neurotoxicity; null mutation; parent grant; programs; protein TDP-43; protein aggregation; single cell analysis; transcriptome sequencing; transcriptomics

Project Summary Frontotemporal dementia (FTD) is an early onset neurodegenerative disease, and the second most common cause of dementia in patients 60 years or younger. The majority of familial FTD are caused by intronic hexanucleotide (CCCCGG) repeat expansion in chromosome 9 open reading frame 72 (C9orf72) gene and by dominant mutations in the Progranulin (GRN) gene, which account for 25% and 15% of familial FTD cases, respectively. These mutations cause haploinsufficiency in both genes and lead to abnormal protein aggregation involving RNA binding protein TDP-43 in neuronal nuclei and cytoplasm. The goal of the parent grant (R01 AA027074-03) is to test the hypothesis that loss of PGRN disrupts neuroimmune interaction in the thalamo- cortical circuit in Grn-/- mice. The purpose of this Diversity Supplement is to expand the scope of the parent grant and characterize the potential interaction of C9orf72 and progranulin in neurodegeneration. To this end, the proposed trainee, Naznin Jahan – a 4th year graduate student in the BMS Graduate Program at UCSF, has established an aging cohort of C9orf72-/-, Grn-/-, and C9orf72-/-;Grn-/- double KO (DKO) mice. The trainee’s results showed that C9orf72-/-;Grn-/- DKO mice exhibit age-dependent neuroinflammation and neuronal loss that are more pronounced than those seen in C9orf72-/- and Grn-/- mice. These findings support the intriguing hypothesis that simultaneous loss-of-function (LOF) in C9orf72 and GRN genes synergistically disrupts glial homeostasis and promote neuronal degeneration in an age dependent manner. The scope of this Diversity Supplement includes (1) to determine the transcriptomic changes regulated by C9orf72 and Grn in glia-neuron homeostasis, and (2) to expand the transcriptomic data using in situ hybridization (ISH), immunohistochemistry and Western blots. In addition, this Diversity Supplement includes a well-defined 2-year timeline, a detailed Mentoring Plan, and Individual Career Development Plan (ICDP) that will significantly enhance the candidate’s research capabilities and complete her dissertation work on the genetic interactions that cause the age dependent neurodegeneration in mice. Working within the proposed timeline, this supplement will prepare the candidate for her long-term career goal as an academic scientist in the field of neuroimmune interaction and neurodegeneration.

项目摘要 额颞叶痴呆（FTD）是一种早发性神经退行性疾病， 老年痴呆症的病因在60岁或更年轻的患者。大多数家族性FTD是由内含子引起的， 在9号染色体开放阅读框72（C9 orf 72）基因中六核苷酸（CCCCGG）重复扩增， 颗粒蛋白原（GRN）基因的显性突变，分别占家族性FTD病例的25%和15%， 分别这些突变导致两个基因的单倍不足，并导致异常的蛋白质聚集 涉及神经元核和细胞质中的RNA结合蛋白TDP-43。父母补助金的目标（R 01 AA 027074 -03）是为了检验PGRN的缺失破坏丘脑中的神经免疫相互作用的假设， Grn-/-小鼠的皮质回路。本多样性补充的目的是扩大母公司的范围 授予和表征C9 orf 72和颗粒蛋白前体在神经变性中的潜在相互作用。为此目的， 拟议的受训者，纳兹宁贾汉-在BMS研究生课程在加州大学旧金山分校的四年级研究生， 建立了C9 orf 72-/-、Grn-/-和C9 orf 72-/-;Grn-/-双KO（DKO）小鼠的老化队列。学员的成绩 显示C9 orf 72-/-;Grn-/- DKO小鼠表现出年龄依赖性神经炎症和神经元损失， 比在C9 orf 72-/-和Grn-/-小鼠中观察到的更明显。这些发现支持了一个有趣的假设 C9 orf 72和GRN基因的同时功能丧失（LOF）协同破坏胶质细胞的稳态， 并以年龄依赖性方式促进神经元变性。本多元化补充协议的范围 包括（1）确定在胶质-神经元稳态中由C9 orf 72和Grn调节的转录组学变化， （2）利用原位杂交（ISH）、免疫组织化学和Western印迹技术， 污点。此外，该多元化补充材料还包括明确的2年时间轴、详细的指导计划、 计划和个人职业发展计划（ICDP），这将大大提高候选人的研究 能力，并完成她的论文工作的遗传相互作用，导致年龄依赖 小鼠的神经变性。在拟议的时间轴内，本补充材料将帮助候选人做好以下准备： 她作为神经免疫相互作用领域的学术科学家的长期职业目标， 神经变性