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Investigating a neuroprotective role of GBA in astrocytes

研究 GBA 对星形胶质细胞的神经保护作用

基本信息

批准号：
10452334
负责人：
Marie Ynez Davis
金额：
$ 19.61万
依托单位：
SEATTLE INST FOR BIOMEDICAL/CLINICAL RES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10452334
关键词：
Age Astrocytes Biological Assay Brain CRISPR/Cas technology Cell Line Cell Survival Cells Chimeric Proteins Coculture Techniques Data Disease Disease Progression Drosophila genus Endocytosis Extracellular Matrix Foundations Genes Human Impairment Individual Infection Investigation Lentivirus Lewy Bodies Lewy body pathology Mediating Modeling Mutation Nerve Degeneration Nervous system structure Neurodegenerative Disorders Neurons Parkinson Disease Pathogenesis Pathogenicity Patients Proteins Regulation Role System Testing Tissues Work alpha synuclein biobank cell type disorder control exosome experimental study extracellular vesicles fly genetic risk factor immunohistochemical markers improved induced pluripotent stem cell innovation mutant neuroprotection new therapeutic target novel novel therapeutic intervention null mutation protein aggregation sex stressor synuclein trafficking uptake

项目摘要

A key feature of Parkinson’s disease (PD) is abnormal protein aggregation within neurons in the brain. These neuronal protein aggregates spread throughout the nervous system as PD progresses, but how this occurs remains unclear. Recent work suggests that astrocytes, which support neuronal function by providing energy and protection from cellular stressors, may also have a neuroprotective role in PD and other neurodegenerative diseases. This proposal investigates a potential new role for the gene GBA, a common genetic risk factor for PD that is also associated with faster disease progression. While most studies of GBA focus exclusively on its role in neurons, we will test whether GBA is required in astrocytes to uptake and degrade neuronal-derived proteins in the extracellular matrix to reduce propagation of pathogenic neuronal protein aggregates. Our recent work using our Drosophila GBA deficient model (GBAdel) of PD indicates that exosomes are an important vehicle mediating the spread of protein aggregation. GBA deficiency dysregulates exosome formation, fusion, and/or content. Surprisingly, we found that glial expression of wildtype GBA can reduce protein aggregation present in GBAdel fly brains. To further investigate a possible neuroprotective role for GBA in astrocytes, we will use cultured neurons and astrocytes differentiated from human induced pluripotent stem cells generated from PD patients carrying GBA mutations. Unrelated healthy age- and sex-matched control and isogenic controls generated by CRISPR/Cas9 reversion of the GBA mutation to wildtype will be used as controls. We will first determine whether GBA is important for astrocyte uptake and processing of neuronal-derived EVs by examining GBA PD versus control astrocyte uptake and intracellular trafficking of neuronal EVs containing synuclein-GFP fusion protein. We will then test whether GBA has a neuroprotective role in astrocytes by co- culturing GBA PD or control astrocytes with GBA PD or control neurons and examine protein aggregation, endolysosomal trafficking and cell survival in both cell types. The results from this study could uncover new therapeutic targets to enhance the neuroprotective function of astrocytes, leading to treatments that could slow or halt the progression of PD and other neurodegenerative diseases characterized by protein aggregates.

帕金森病 (PD) 的一个关键特征是大脑神经元内蛋白质异常聚集。随着帕金森病的进展，这些神经元蛋白聚集体会扩散到整个神经系统，但是这种现象是如何发生的呢？发生情况仍不清楚。最近的研究表明，星形胶质细胞通过提供支持神经元功能能量和免受细胞应激源的保护，也可能在帕金森病和其他疾病中具有神经保护作用神经退行性疾病。该提案研究了基因 GBA 的潜在新作用，GBA 是一种常见的基因。 PD 的遗传风险因素也与疾病进展更快有关。虽然大多数大湾区研究专注于其在神经元中的作用，我们将测试星形胶质细胞是否需要 GBA 来摄取和降解细胞外基质中的神经元衍生蛋白，以减少致病性神经元的传播蛋白质聚集体。我们最近使用果蝇 GBA 缺陷模型 (GBAdel) 进行的 PD 研究表明，外泌体是一种介导蛋白质聚集传播的重要载体。 GBA 缺乏会导致外泌体形成失调，融合和/或内容。令人惊讶的是，我们发现野生型GBA的神经胶质表达可以减少蛋白质 GBAdel 果蝇大脑中存在聚集。进一步研究 GBA 可能的神经保护作用星形胶质细胞，我们将使用培养的神经元和由人类诱导多能干细胞分化而来的星形胶质细胞由携带 GBA 突变的 PD 患者产生。不相关的健康年龄和性别匹配控制和通过 CRISPR/Cas9 将 GBA 突变恢复为野生型而生成的等基因对照将用作对照。我们将首先确定 GBA 对于星形胶质细胞的摄取和神经元来源的 EV 的处理是否重要通过检查 GBA PD 与对照星形胶质细胞的摄取和神经元 EV 的细胞内运输，其中含有突触核蛋白-GFP 融合蛋白。然后我们将通过共同测试 GBA 是否对星形胶质细胞具有神经保护作用培养 GBA PD 或对照星形胶质细胞与 GBA PD 或对照神经元并检查蛋白质聚集，两种细胞类型的内溶酶体运输和细胞存活。这项研究的结果可能会发现新的治疗目标是增强星形胶质细胞的神经保护功能，从而导致治疗可能减缓或阻止帕金森病和其他以蛋白质聚集为特征的神经退行性疾病的进展。