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

项目摘要 额颞性痴呆(FTD)是一种早期发病的神经退行性疾病，也是第二常见的 60岁或以下患者患痴呆症的原因。大多数家族性FTD是由内含子引起的 染色体9开放阅读框72(C9orf72)基因中六核苷酸(CCCCGG)重复序列的扩增 原颗粒蛋白(GRN)基因的显性突变，分别占家族性FTD病例的25%和15%， 分别进行了分析。这些突变导致两个基因的单倍性不足，并导致蛋白质异常聚集。 RNA结合蛋白TDP-43位于神经元的胞核和胞浆中。家长资助的目标(R01 AA027074-03)是为了测试PGRN丢失扰乱丘脑-大脑中神经免疫相互作用的假说。 GRN-/-小鼠的皮质环路。本多样性补充文件的目的是扩大母公司的范围 授予并描述C9orf72和原颗粒蛋白在神经退行性变中的潜在相互作用。为此， 拟议的实习生纳兹宁·贾汉是加州大学旧金山分校BMS研究生项目的四年级研究生，他拥有 建立了C9orf72-/-、Grn-/-和C9orf72-/-；Grn-/-Double KO(DKO)小鼠的衰老队列。实习生的成绩 显示C9orf72-/-；Grn-/-DKO小鼠表现出年龄依赖性的神经炎症和神经元丢失 比C9orf72-/-和Grn-/-小鼠更明显。这些发现支持这一耐人寻味的假设 C9orf72和GRN基因同时功能丧失(LOF)协同破坏神经胶质细胞内稳态 并以一种年龄依赖的方式促进神经元退化。本多样性补充文件的范围 包括(1)确定C9orf72和Grn在胶质细胞-神经元稳态中调控的转录变化， (2)利用原位杂交、免疫组织化学和免疫印迹技术对转录数据进行扩展 斑点。此外，这份多样性补充资料还包括一个明确的两年时间表，一个详细的指导 计划和个人职业发展计划(ICDP)，这将显著加强候选人的研究 能力，并完成她关于导致年龄依赖的遗传相互作用的论文工作 小鼠的神经变性。在建议的时间表内，本附录将为候选人做好准备 她的长期职业目标是成为神经免疫相互作用和领域的学术科学家 神经退行性变。

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.