N-terminal huntingtin and Huntington disease neuropathology

N 末端亨廷顿蛋白和亨廷顿病神经病理学

基本信息

批准号：
10117290
负责人：
Andrew P Escayg
金额：
$ 44.84万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-03-01 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10117290
关键词：
Address Adult Affect Age Amino Acids Animals Antisense Oligonucleotides Brain Brain Diseases Brain region CRISPR/Cas technology Cell Nucleus Cells Conserved Sequence Corpus striatum structure Development Disease Embryo Embryonic Development Event Excision Gene Targeting Genes Genetic Transcription Genets Guide RNA Huntington Disease Huntington gene Huntington protein Impairment Injections Intervention Knock-in Mouse Knockout Mice Lead Length Mediating Methods Motor Mus Mutation N-terminal Nerve Degeneration Neurodegenerative Disorders Neurologic Neurons Nuclear Pathogenesis Pathologic Pathology Patients Phenotype Proteins Proteolysis Small Interfering RNA Testing Therapeutic Therapeutic Effect Toxic effect age related experimental study genome editing inhibitor/antagonist insight mutant neuronal survival neuropathology neurotoxic novel therapeutic intervention offspring polyglutamine preservation protective effect side effect therapeutically effective tool treatment strategy

项目摘要

Huntington's disease (HD) is a devastating neurodegenerative disease caused by expansion of a polyglutamine (polyQ) domain in distinct proteins with different functions. In HD, the polyQ domain is located in the N-terminal region of huntingtin (Htt). This N-terminal region is well conserved in a wide range of species, but polyQ expansion can lead to misfolding and subsequent toxicity of N-terminal fragments of Htt. Since a lack of Htt causes embryonic lethality in mice, Htt is also thought to be essential for animal development and survival. Reducing the expression of mutant Htt is widely accepted as an important strategy for treating HD, so considerable efforts have gone into developing siRNA and anti-sense oligonucleotides to suppress the expression of mutant Htt. These approaches have also raised concerns that markedly suppressing Htt expression could lead to side effects by diminishing the normal function of Htt; however, whether Htt can preserve critical functions without the N-terminal domain that contains the polyQ domain remains unknown. Addressing this issue is important if we are to develop a new strategy to treat HD: if the N-terminal polyQ domain is not required for essential Htt functions and can be removed, complete elimination of the N-terminal region of Htt is now possible since the recent development of the genomic editing tool, CRISPR/Cas9. In this competitive renewal application, we will use CRISPR/Cas9 to investigate the toxicity of N-terminal mutant Htt fragments and therapeutic effects by removing the polyQ-containing N-terminal region. In Aim 1, we will use CRISPR/Cas9 to introduce mutations in the mouse Htt gene in embryos from HD 140Q KI mice to generate truncated mutant Htt genes that express different N-terminal Htt fragments and can be transmitted to offspring via the germline. Using the newly established HD KI mice that express different N-terminal mHtt fragments containing the same polyQ repeat (140Q) at the endogenous level, we will examine the relationship between the length of N-terminal mutant Htt fragments and their nuclear accumulation and toxicity in striatal neurons. In Aim 2, we will use CRISPR/Cas9 to remove the N-terminal polyQ domain as a therapeutic strategy. We will explore whether removing the N-terminal polyQ domain in Htt can eliminate neuropathology without affecting neuronal survival and function in adult mice. These studies will use HD knock-in mice in which mutant Htt is expressed at the same endogenous level as in HD patients. We hope these studies will not only provide new insight into the pathogenesis of N-terminal mutant Htt fragments, but also allow us to develop a novel therapeutic strategy to treat Huntington's disease and other polyQ diseases.

亨廷顿舞蹈症 (HD) 是一种毁灭性的神经退行性疾病，由神经细胞扩张引起具有不同功能的不同蛋白质中的聚谷氨酰胺 (polyQ) 结构域。在 HD 中，polyQ 域位于亨廷顿蛋白 (Htt) 的 N 末端区域。这个 N 末端区域在多种物种中都得到了很好的保存，但 PolyQ 扩展可能导致 Htt N 末端片段的错误折叠和随后的毒性。自从一个缺乏 Htt 会导致小鼠胚胎死亡，Htt 也被认为对于动物发育和生存。减少突变型Htt的表达被广泛认为是治疗HD的重要策略，因此人们付出了巨大的努力来开发 siRNA 和反义寡核苷酸来抑制突变体Htt的表达。这些方法也引起了人们的担忧，即显着抑制 Htt 表达可能会削弱 Htt 的正常功能，从而导致副作用；然而，Htt是否可以在没有包含 PolyQ 结构域的 N 端结构域的情况下保留关键功能仍然未知。如果我们要开发治疗 HD 的新策略，解决这个问题就很重要：如果 N 端 polyQ 结构域对于基本的 Htt 功能来说不是必需的，可以删除，完全消除 N 端自从最近开发了基因组编辑工具 CRISPR/Cas9 以来，Htt 区域现在已成为可能。在这个竞争性更新申请，我们将使用CRISPR/Cas9研究N端突变体Htt的毒性通过去除含有polyQ的N端区域来观察片段和治疗效果。在目标 1 中，我们将使用 CRISPR/Cas9 在 HD 140Q KI 小鼠胚胎中引入小鼠 Htt 基因突变，以产生截短的突变型 Htt 基因，表达不同的 N 端 Htt 片段，并且可以遗传给后代通过种系。使用新建立的表达不同N端mHtt片段的HD KI小鼠在内源水平上包含相同的polyQ重复序列（140Q），我们将检查之间的关系 N 端突变 Htt 片段的长度及其在纹状体神经元中的核积累和毒性。在目标 2，我们将使用 CRISPR/Cas9 去除 N 端 polyQ 结构域作为治疗策略。我们将探讨去除 Htt 中的 N 端 polyQ 结构域是否可以消除神经病理学而不影响成年小鼠的神经元存活和功能。这些研究将使用 HD 敲入小鼠，其中突变型 Htt 是其内源性表达水平与 HD 患者相同。我们希望这些研究不仅能够提供新的深入了解 N 端突变 Htt 片段的发病机制，也使我们能够开发出一种新的治疗亨廷顿病和其他 PolyQ 疾病的治疗策略。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A Huntingtin Knockin Pig Model Recapitulates Features of Selective Neurodegeneration in Huntington's Disease.

亨廷顿基因敲入猪模型概括了亨廷顿病选择性神经变性的特征。

DOI：
发表时间：
2018
期刊：
影响因子：
64.5
作者：
Yan, Sen;Tu, Zhuchi;Liu, Zhaoming;Fan, Nana;Yang, Huiming;Yang, Su;Yang, Weili;Zhao, Yu;Ouyang, Zhen;Lai, Chengdan;Yang, Huaqiang;Li, Li;Liu, Qishuai;Shi, Hui;Xu, Guangqing;Zhao, Heng;Wei, Hongjiang;Pei, Zhong;Li, Shihua;Lai, Liangxue
通讯作者：
Lai, Liangxue

HAP1 Is Required for Endocytosis and Signalling of BDNF and Its Receptors in Neurons.

HAP1 是神经元中 BDNF 及其受体的内吞作用和信号传导所必需的。

DOI：
发表时间：
2018
期刊：
影响因子：
5.1
作者：
Lim, Yoon;Wu, Linda Lin;Chen, Si;Sun, Ying;Vijayaraj, Swarna Lekha;Yang, Miao;Bobrovskaya, Larisa;Keating, Damien;Li, Xiao;Zhou, Xin
通讯作者：
Zhou, Xin

Cerebellum-enriched protein INPP5A contributes to selective neuropathology in mouse model of spinocerebellar ataxias type 17.

小脑富含蛋白 INPP5A 有助于 17 型脊髓小脑共济失调小鼠模型的选择性神经病理学。

DOI：
发表时间：
2020
期刊：
影响因子：
16.6
作者：
Liu, Qiong;Huang, Shanshan;Yin, Peng;Yang, Su;Zhang, Jennifer;Jing, Liang;Cheng, Siying;Tang, Beisha;Li, Xiao;Pan, Yongcheng;Li, Shihua
通讯作者：
Li, Shihua

Synergistic Toxicity of Polyglutamine-Expanded TATA-Binding Protein in Glia and Neuronal Cells: Therapeutic Implications for Spinocerebellar Ataxia 17.

神经胶质细胞和神经元细胞中聚谷氨酰胺扩展的 TATA 结合蛋白的协同毒性：脊髓小脑性共济失调的治疗意义 17。