TDP-43 acetylation, phase separation, aggregation, and clearance by antibody-mediated degradation

TDP-43 乙酰化、相分离、聚集和抗体介导的降解清除

• 批准号: 10184466
• 负责人: GEOFFREY A CHANG
• 金额: $ 75.92万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10184466
• 关键词: ALS patients; ATP phosphohydrolase; Acetylation; Acetyltransferase; Aging; Alzheimer' s Disease; Alzheimer' s disease patient; Amyotrophic Lateral Sclerosis; Antibodies; Automobile Driving; Autopsy; Binding; Biological; Biology; Brain; Cell model; Complex; Cytoplasm; Deacetylase; Deacetylation; Dementia; Disease; EP300 gene; Elderly; Engineering; Enzymes; Excision; Exhibits; Frontotemporal Dementia; Genes; Heat-Shock Proteins 70; Hippocampus (Brain); Histone Deacetylase; Human; Link; Liquid substance; Mediating; Methods; Modification; Molecular Chaperones; Motor Neurons; Mutation; Names; Neuraxis; Neurodegenerative Disorders; Neurons; Nuclear; Outcome; Pathologic; Pathology; Patients; Periodicity; Phase; Phase Transition; Property; RNA Binding; RNA Processing; RNA Splicing; RNA-Binding Proteins; Regulation; Reporting; Rodent Model; Role; Seeds; Spinal Cord; Stress; Syndrome; TDP-43 aggregation; Therapeutic; Transcription Initiation; age related; aged; induced pluripotent stem cell; inhibitor/antagonist; insight; limbic-predominant age-related TDP-43 encephalopathy; mouse model; multicatalytic endopeptidase complex; mutant; nanobodies; neuron loss; neurotoxicity; prevent; protein TDP-43; stress granule; transcriptome; ubiquitin-protein ligase

Cytoplasmic aggregation of TDP-43 has been reported in almost every age-dependent neurodegenerative disease, including in >40% of frontal temporal dementia (FTD), in the hippocampal neurons of Alzheimer's disease (AD) patients, in >90% of ALS, and in ~100% of a recently recognized AD-like dementia in the oldest of the elderly, an AD-like syndrome identified in 2019 and named Limbic-predominant Age-related TDP-43 Encephalopathy (LATE). We have demonstrated that TDP-43 phase separation and aggregation can drive neuronal death independent of RNA binding, stress granule formation, and TDP-43 association with stress granules. We have subsequently identified that acetylation of TDP-43 (which abolishes its RNA binding) drives its separation into liquid spherical annular bodies. These nuclear annuli have liquid annular shells enriched in TDP-43 and liquid centers highly enriched in HSP70 family molecular chaperones. Use of inhibitors of known deacetylases or the proteasome (to mimic the known age-dependent declines in deacetylase and proteasome activities) provokes cytoplasmic TDP-43 aggregation. We propose to determine the biological and pathological role(s) of acetylated TDP-43 and how HSP70 chaperone activity regulates nuclear TDP-43 function and its aggregation in the cytoplasm. We will determine the regulation and biological consequences of acetylated TDP-43 in neurons by identifying the key regulatory enzymes (acetyltranferases and deacetylases) of acetylated TDP-43 and alter TDP-43’s function in RNA splicing and its subcellular localization/aggregation. To understand how HSP70 family molecular chaperones regulates phase transition of TDP-43, we will use Hsc70 (encoded by the HSPA8 gene and the most abundant HSP70 in neurons) and determine how Hsc70 interacts with TDP-43. We will also determine if enhancing the activity of HSP70 (such as HSPA8, which is highly expressed in neurons) ameliorates TDP-43 pathology. We will also develop a potential therapeutic approach for TDP-43 proteinopathies in which rapid proteasome-mediated degradation of aggregated TDP-43 is achieved through an engineered E3 ubiquitin ligase linked to a synthetically evolved nanobody (a single chain antibody derived from an antibody heavy chain) recognizing either acetylated or phosphorylated TDP-43. Outcomes of these efforts will provide key insights for understanding basic aspects of TDP-43 biology and pathobiology in common dementia, and for developing a new concept of therapy that specifically targets TDP- 43 pathology that could potentially benefit aged patients with TDP-43-related dementia.

在几乎所有依赖年龄的神经退行性中，TDP-43的细胞质聚集已报道 在阿尔茨海默氏症的海马神经元中，疾病，包括40％的额叶临时痴呆（FTD）中的疾病 疾病（AD）患者，> 90％的ALS，在最古老的最近公认的广告样痴呆症中约有100％ 在2019年确定的广告状综合征，并命名为与年龄相关的TDP-43 脑病（晚期）。我们已经证明了TDP-43相分离和聚集可以驱动 神经元死亡独立于RNA结合，应力颗粒形成和TDP-43与应力的关联 颗粒。随后，我们确定了TDP-43的乙酰化（废除其RNA结合） 将其分离为液体球形环体。这些核环具有液态环形壳 富含TDP-43和液体中心，高度富含HSP70家族分子伴侣。使用 已知脱乙酰基酶或蛋白酶体的抑制剂（模仿已知年龄依赖性脱乙酰基酶的下降 和蛋白酶体活动）挑衅细胞质TDP-43聚集。我们建议确定生物学 乙酰化TDP-43的病理作用以及HSP70伴侣活性如何调节核TDP-43 功能及其在细胞质中的聚集。我们将确定调节和生物学后果 通过鉴定关键调节酶（乙酰基糖酶和脱乙酰基酶），神经元中的乙酰化TDP-43 乙酰化的TDP-43和改变TDP-43在RNA剪接及其亚细胞定位/聚集中的功能。 要了解HSP70家族分子伴侣如何调节TDP-43的相变，我们将使用HSC70 （由HSPA8基因和神经元中最丰富的HSP70编码）并确定HSC70如何相互作用 与TDP-43。我们还将确定是否增强了HSP70的活性（例如HSPA8，这是高度的 在神经元中表达）改善TDP-43病理学。我们还将开发一种潜在的治疗方法 TDP-43蛋白质病，其中快速蛋白酶体介导的聚集TDP-43降解 通过与合成进化的纳米病（单链）相关的工程E3泛素连接酶 识别乙酰化或磷酸化的TDP-43的抗体重链的抗体。 这些努力的结果将为理解TDP-43生物学的基本方面和 普通痴呆症的病理生物学，以及开发一种新的治疗概念，该概念专门针对TDP- 43可能使TDP-43相关痴呆患者老年患者受益的病理。