An optogenetic approach to establish the interplay between stress granules and TDP-43 proteinopathy in ALS/FTD

一种光遗传学方法来确定 ALS/FTD 中应激颗粒与 TDP-43 蛋白病之间的相互作用

• 批准号: 10370344
• 负责人: Christopher James Donnelly
• 金额: $ 37.22万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-15 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370344
• 关键词: ALS patients; Address; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Autopsy; Biological Models; Brain; Cell Nucleus; Cell physiology; Cells; Cellular Stress; Chronic; Chronic stress; Cytoplasm; Cytoplasmic Inclusion; DNA Sequence Alteration; DNA-Binding Proteins; Development; Diagnosis; Disease; Exhibits; Family; Family history of; Frontotemporal Dementia; Functional disorder; Genes; Genetic; Heterogeneity; Homeostasis; Human; Impairment; Interneurons; Light; Link; Literature; Mediating; Methods; Modeling; Motor Neurons; Mutation; Nervous system structure; Neurodegenerative Disorders; Neurons; Nuclear RNA; Onset of illness; Organelles; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Patients; Play; Process; Protein Biosynthesis; Protein Conformation; Proteins; RNA Processing; RNA interference screen; Recording of previous events; Regulation; Seeds; Spinal Cord; System; Tissues; Toxic effect; chronic traumatic encephalopathy; cytotoxic; experimental study; extracellular; frontotemporal lobar dementia-amyotrophic lateral sclerosis; functional loss; genome-wide; induced pluripotent stem cell; insight; motor neuron degeneration; mutant; nervous system disorder; neuropathology; neurotoxic; neurotoxicity; novel; optogenetics; overexpression; prevent; protein TDP-43; sporadic amyotrophic lateral sclerosis; stress granule; stressor

Amyotrophic lateral sclerosis (ALS) is a progressive, fatal neurodegenerative disorder marked by loss of motor neurons of the spinal cord and cortex. ALS is characterized by significant heterogeneity in disease onset and patient presentation. Over the last twenty years, mutations in over 35 different genes have been uncovered as causal in the development of familial forms of ALS (fALS); however, fALS only accounts for roughly 10% of all ALS cases. The remaining 90% of patients suffer from sporadic ALS (sALS), with no family history of disease and unknown causes of pathogenesis. Regardless of all this genetic and pathogenic complexity, remarkably nearly every single ALS patient (both fALS and sALS) share a common neuropathology in the form of aberrant cytoplasmic inclusions of a protein called TAR DNA-binding protein of 43 kDa (TDP-43) found in degenerating regions of the nervous system. Insight drawn from studies of fALS-linked mutations have implicated stress granule dysfunction in disease development, but the exact mechanisms by which the stress granules may directly regulate TDP-43 aggregation remains unclear. By utilizing a novel optogenetic approach to induce either TDP-43 inclusions or functional stress granules, we can manipulate these processes at a previously unattainable level of control. These experiments will explore the contribution of stress granule dysfunction to TDP-43 inclusion neurotoxicity. We hypothesize that perturbations in stress granule dynamics contribute to neurotoxic TDP-43 aggregation in ALS. We will first investigate the contribution of stress granules to light-induced TDP-43 inclusions. We will then determine if chronic or persistent stress granule formation initiates TDP-43 proteinopathy. Finally, we will perform a genome-wide RNAi screen to identify novel pathways that protect against TDP-43 inclusion toxicity and assess whether this protection is dependent on stress granule regulation.

肌萎缩侧索硬化症 (ALS) 是一种进行性、致命性的神经退行性疾病，其特征是 由于脊髓和皮质运动神经元的丧失。 ALS 的特点是显着 疾病发作和患者表现的异质性。过去二十年来，突变 超过 35 个不同的基因已被发现与家族形式的发展有关 肌萎缩侧索硬化症（fALS）；然而，fALS 仅占所有 ALS 病例的大约 10%。其余 90%的患者患有散发性ALS（sALS），无家族病史且未知 发病原因。尽管存在所有这些遗传和致病的复杂性，但值得注意的是 几乎每一位 ALS 患者（fALS 和 sALS）都有共同的神经病理学特征 43 kDa 的 TAR DNA 结合蛋白的异常细胞质内含物形式 (TDP-43) 存在于神经系统的退化区域。从研究中得出的见解 fALS 相关突变与疾病发展中的应激颗粒功能障碍有关，但是 应激颗粒直接调节 TDP-43 聚集的确切机制 仍不清楚。通过利用新型光遗传学方法诱导 TDP-43 内含物或 功能性应激颗粒，我们可以以以前无法实现的方式操纵这些过程 控制水平。这些实验将探讨应激颗粒功能障碍对 TDP-43包合物的神经毒性。我们假设应力颗粒动力学的扰动 有助于 ALS 中神经毒性 TDP-43 的聚集。我们首先要调查的贡献 应力颗粒对光诱导的 TDP-43 夹杂物的影响。然后我们将确定是否是慢性或 持续的应激颗粒形成引发 TDP-43 蛋白病。最后，我们将执行一个 全基因组 RNAi 筛选，以确定防止 TDP-43 包含的新途径 毒性并评估这种保护是否依赖于应激颗粒调节。

项目成果

Optogenetic TDP-43 nucleation induces persistent insoluble species and progressive motor dysfunction in vivo.

光遗传学 TDP-43 成核可诱导持久的不溶性物质和进行性体内运动功能障碍。

• 发表时间: 2020
• 影响因子: 6.1
• 作者: Otte, Charlton G;Fortuna, Tyler R;Mann, Jacob R;Gleixner, Amanda M;Ramesh, Nandini;Pyles, Noah J;Pandey, Udai B;Donnelly, Christopher J
• 通讯作者: Donnelly, Christopher J

Disrupting pathologic phase transitions in neurodegeneration.

破坏神经变性的病理相变。

• 发表时间: 2023-07-03
• 作者: Hurtle, Bryan T;Xie, Longxin;Donnelly, Christopher J
• 通讯作者: Donnelly, Christopher J

Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy.

FG-核孔蛋白招募核输入受体来挽救 TDP-43 蛋白病的特征。

• 发表时间: 2022-12-08
• 影响因子: 15.1
• 作者: Khalil, Bilal;Chhangani, Deepak;Wren, Melissa C;Smith, Courtney L;Lee, Jannifer H;Li, Xingli;Puttinger, Christian;Tsai, Chih;Fortin, Gael;Morderer, Dmytro;Gao, Junli;Liu, Feilin;Lim, Chun Kim;Chen, Jingjiao;Chou, Ching;Croft, Cara
• 通讯作者: Croft, Cara

NUP62 localizes to ALS/FTLD pathological assemblies and contributes to TDP-43 insolubility.

NUP62 定位于 ALS/FTLD 病理组装并导致 TDP-43 不溶。

• 发表时间: 2022-06-13
• 影响因子: 16.6
• 作者: Gleixner, Amanda M;Verdone, Brandie Morris;Otte, Charlton G;Anderson, Eric N;Ramesh, Nandini;Shapiro, Olivia R;Gale, Jenna R;Mauna, Jocelyn C;Mann, Jacob R;Copley, Katie E;Daley, Elizabeth L;Ortega, Juan A;Cicardi, Maria Elena;Kiskinis, Evang
• 通讯作者: Kiskinis, Evang

DDX17 is involved in DNA damage repair and modifies FUS toxicity in an RGG-domain dependent manner.

DDX17 参与 DNA 损伤修复，并以 RGG 结构域依赖性方式改变 FUS 毒性。

• 发表时间: 2021
• 影响因子: 12.7
• 作者: Fortuna, Tyler R;Kour, Sukhleen;Anderson, Eric N;Ward, Caroline;Rajasundaram, Dhivyaa;Donnelly, Christopher J;Hermann, Andreas;Wyne, Hala;Shewmaker, Frank;Pandey, Udai Bhan
• 通讯作者: Pandey, Udai Bhan