Mechanisms of autophagic dysfunction in progranulin-related neurodegeneration

颗粒体蛋白前体相关神经变性中自噬功能障碍的机制

基本信息

批准号：
10327305
负责人：
Michael Fernandopulle
金额：
$ 5.18万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-12-01 至 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10327305
关键词：
Address Affect Affinity Aging Amyotrophic Lateral Sclerosis Annexins Area Autophagocytosis Autophagosome Axon Binding Binding Proteins Binding Sites Biochemical Blood Brain Diseases Calcium Calcium Channel Agonists Capsid Proteins Cell division Cells Characteristics Chimeric Proteins Co-Immunoprecipitations Complex Data Defect Degenerative Disorder Dementia Dendrites Disease Dissection Endoplasmic Reticulum Fluorescence Resonance Energy Transfer Fluorescent Probes Frontotemporal Dementia Functional disorder Gene Mutation Generations Genes Genetic Glycoproteins Homeostasis Impairment Incidence Induced pluripotent stem cell derived neurons Infection Label Lead Lysosomes Mediating Medical Genetics Membrane Microscopy Mitotic Molecular Monitor Mutation Nerve Degeneration Nervous system structure Neurocognitive Neurodegenerative Disorders Neuromuscular Diseases Neurons Organelles PGRN gene Pathologic Pathway interactions Patients Process Protein Biosynthesis Protein Export Pathway Proteins Proteomics Quality Control Reactive Oxygen Species Research Site Skin Starvation Structure Synapses Techniques Testing Time Work age related neurodegeneration base cell age cell injury dementia risk effective therapy familial amyotrophic lateral sclerosis frontotemporal lobar dementia-amyotrophic lateral sclerosis functional loss genetic manipulation inhibitor loss of function mutation member misfolded protein new therapeutic target overexpression oxidative damage prevent protein misfolding recruit stem cells stressor

项目摘要

PROJECT SUMMARY As cells age, protein quality control becomes increasingly important. Accumulated stressors such as starvation, oxidative damage, and infections lead to organelle dysfunction and protein misfolding. Stem cells, such as those present in the skin, gut, and blood, can dilute these insults through cell division. Neurons and other post-mitotic cells, however, must confront them directly. Autophagy, or lysosome-mediated degradation, is the primary mechanism for clearing large dysfunctional entities within the cell. Neuronal autophagy must be especially robust for two main reasons: 1) the high rates of protein synthesis and ATP generation in neurons entail a higher incidence of misfolded proteins and reactive oxygen species, both of which damage the cell, and 2) high spatial separation of multiple specialized regions (e.g. axons, dendrites, synapses) demands local clearance of damage in those areas to prevent key functional loss. Predictably, errors in autophagy often affect the aging nervous system. Frontotemporal dementia (FTD), a progressive neurocognitive disease, is associated with several single-gene mutations involved in autophagy and broader protein quality control. Many of these genes overlap with those implicated in amyotrophic lateral sclerosis (ALS), a common neuromuscular disease. Progranulin (PGRN), a monogenic cause of FTD and risk modifier for ALS, is a lysosomal glycoprotein that causes defects in autophagy through an unknown mechanism. Recent work by our lab has demonstrated that another ALS- associated protein, annexin A11 (ANXA11), shows decreased recruitment to the lysosome in PGRN deficient neurons. ER exit site (ERES) proteins, which regulate protein export from the endoplasmic reticulum, also show decreased lysosomal recruitment in PGRN deficiency. ANXA11 is known to bind members of ERESs, as well as to associate physically with the lysosome. Furthermore, ERESs may be involved in more than just protein export. Our collaborators recently discovered a phenomenon where lysosomes directly engulf and degrade ERESs bearing misfolded proteins—a clear example of autophagic involvement. I propose to test the hypothesis that PGRN regulates ERES autophagy through ANXA11 action by addressing the following specific aims: 1) determine how PGRN regulates ANXA11 interaction with lysosome-organelle contact sites in iPSC-derived neurons, and 2) determine how PGRN and ANXA11 jointly regulate autophagic activity. I will use a combination of sophisticated microscopy techniques, biochemical protein identification, and targeted genetic manipulation to complete these aims. Uncovering the mechanism of PGRN-related neurodegeneration could lead to a better understanding of the shared pathophysiology of FTD and ALS, providing new drug targets for these incurable and universally devastating diseases of aging.

项目摘要随着细胞的年龄，蛋白质质量控制变得越来越重要。累积的压力源，例如饥饿，氧化损伤和感染会导致细胞器功能障碍和蛋白质折叠式折叠。干细胞，例如存在于皮肤，肠道和血液中，可以通过细胞分裂稀释这些侮辱。神经元和其他有丝分裂但是，细胞必须直接面对它们。自噬或溶酶体介导的降解是主要的清除细胞内大功能失调实体的机制。神经元自噬必须特别强大有两个主要原因：1）神经元中蛋白质合成和ATP产生的高速率需要更高错误折叠的蛋白质和活性氧的发生率，两者都会损害细胞，2）高空间分离多个专业区域（例如轴突，树突，突触）需要局部清除损害在这些领域，以防止关键功能损失。可以预见的是，自噬的错误通常会影响衰老的神经系统。额前痴呆（FTD）是一种进行性神经认知疾病，与几种有关单基因突变涉及自噬和更广泛的蛋白质质量控制。这些基因中有许多重叠与肌萎缩性侧索硬化症（ALS）中隐含的那些是一种常见的神经肌肉疾病。前植物（PGRN）是ALS的FTD和风险修饰符的单基因原因，是一种溶酶体糖蛋白，会导致缺陷通过未知机制在自噬中。我们的实验室最近的工作表明，另一个ALS- 相关蛋白质Annexin A11（Anxa11）显示了PGRN默认溶酶体的精制募集神经元。 ER出口位点（ERES）蛋白质（调节内质网的蛋白质出口）也显示 PGRN缺乏症的溶酶体募集降低。众所周知，Anxa11绑定了Eress的成员，以及与溶酶体物理结合。此外，ERESS不仅参与蛋白质出口。我们的合作者最近发现了一种现象，溶酶体直接吞噬并降解ERESS 具有错误折叠的蛋白质 - 自噬参与的明确例子。我建议检验以下假设 PGRN通过解决以下特定目的来调节ERES通过Anxa11动作来调节自噬：1）确定PGRN如何调节Anxa11与IPSC衍生的溶酶体 - 轨道接触位点的相互作用神经元，以及2）确定PGRN和ANXA11如何共同调节自噬活性。我将使用一个组合复杂的显微镜技术，生化蛋白鉴定和靶向遗传操作完成这些目标。发现与PGRN相关的神经变性的机制可能导致更好了解FTD和ALS共享的病理生理学，为这些无法治愈和衰老的普遍毁灭性疾病。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Development and Comparative Evaluation of Endolysosomal Proximity Labeling-Based Proteomic Methods in Human iPSC-Derived Neurons.

DOI：
10.1021/acs.analchem.0c03107
发表时间：
2020-12-01
期刊：
Analytical chemistry
影响因子：
7.4
作者：
Frankenfield AM;Fernandopulle MS;Hasan S;Ward ME;Hao L
通讯作者：
Hao L

Transcription Factor-Mediated Differentiation of Human iPSCs into Neurons.

DOI：
10.1002/cpcb.51
发表时间：
2018-06
期刊：
Current protocols in cell biology
影响因子：
0
作者：
Fernandopulle MS;Prestil R;Grunseich C;Wang C;Gan L;Ward ME
通讯作者：
Ward ME

RNA transport and local translation in neurodevelopmental and neurodegenerative disease.

DOI：
10.1038/s41593-020-00785-2
发表时间：
2021-05
期刊：
Nature neuroscience
影响因子：
25
作者：
Fernandopulle MS;Lippincott-Schwartz J;Ward ME
通讯作者：
Ward ME

SNX19 restricts endolysosome motility through contacts with the endoplasmic reticulum.

DOI：
10.1038/s41467-021-24709-1
发表时间：
2021-07-27
期刊：
Nature communications
影响因子：
16.6
作者：
Saric A;Freeman SA;Williamson CD;Jarnik M;Guardia CM;Fernandopulle MS;Gershlick DC;Bonifacino JS
通讯作者：
Bonifacino JS