Determining the Interacting Effects of GBA, SNCA, and APOE on a-Synuclein Pathology Severity in Dementia with Lewy Bodies and Parkinson's Disease

确定 GBA、SNCA 和 APOE 对路易体痴呆和帕金森病的 a-突触核蛋白病理严重性的相互作用

• 批准号: 10603361
• 负责人: Camille Goldman
• 金额: $ 4.61万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2025-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10603361
• 关键词: Affect; Age; Alzheimer' s Disease; American; Animal Model; Antibodies; Astrocytes; Autophagocytosis; Biochemistry; Biological Assay; Brain; Cell Line; Cell model; Cells; Cerebrosides; Cholesterol; Clinical; Coculture Techniques; Complex; Coupled; Coupling; Dementia with Lewy Bodies; Development; Disease; Disease Progression; Early Diagnosis; Environmental Risk Factor; Enzymes; Functional disorder; Genes; Genetic; Genomics; Genotype; Goals; Health; Heterogeneity; Homeostasis; Human; Image; Knock-out; Knowledge; Lead; Link; Measures; Membrane Potentials; Metabolic; Mitochondria; Modeling; Molecular; Mutation; Nerve Degeneration; Neurobehavioral Manifestations; Neurodegenerative Disorders; Neurons; Outcome; Parkinson Disease; Pathogenicity; Pathologic; Pathology; Pathway interactions; Patients; Persons; Phenotype; Phosphorylation; Population; Prevention; Proteins; Proteomics; Publishing; Reporter; Risk Factors; Severities; Techniques; Technology; Testing; Therapeutic; Tissue Model; United States; Variant; Western Blotting; Work; alpha synuclein; apolipoprotein E-3; apolipoprotein E-4; brain tissue; cell type; dopaminergic neuron; dosage; effective therapy; flexibility; genetic risk factor; genetic variant; immunocytochemistry; induced pluripotent stem cell; insight; knock-down; lipid metabolism; loss of function mutation; motor symptom; new technology; next generation; novel strategies; novel therapeutic intervention; novel therapeutics; prevent; response; single-cell RNA sequencing; synucleinopathy; transcriptomics

Project Summary Synucleinopathies, including dementia with Lewy bodies (DLB) and Parkinson’s disease (PD), are a growing health crisis, affecting over 1 million people in the United States and an estimated 10 million people globally. Synucleinopathies are caused by accumulation of the protein α-synuclein (α-syn) into unnatural fibrils in the brain and can manifest with motor symptoms, cognitive symptoms, or a combination of the two. There are currently no effective treatments to cure or prevent synucleinopathies, in large part because there is no clear cause for disease. There are many identified genetic and environmental risk factors associated with synucleinopathies, but the context in which these factors lead to disease is still unknown. In addition to direct mutations to SNCA, the gene that encodes α-syn, mutations to GBA and APOE are well established risk factors and have been shown to increase α-syn aggregation. However, none of these risk factors are great predictors of disease on their own, suggesting unknown genetic and environmental interactions likely influence the initiation, severity, and clinical outcomes of α-syn pathology. In this proposal, I focus on how both cell type and genotype interact with each other to modulate α-syn pathology severity. I hypothesize that α-syn pathology is precipitated by epistatic interactions between genetic factors that disrupt homeostasis in different cell types and leaves dopaminergic neurons vulnerable to deleterious α-syn pathology and neurodegeneration. I will investigate this hypothesis using two independent, complementary approaches to dissect the cellular and molecular mechanisms that modulate α-syn phenotypes. First, I will couple GBA knockdown with SNCA-triplication, wild-type, and knockout cell lines. There is a clear link between GBA mutations and α-syn aggregation, but how these proteins connect is still unknown. I will use high-content imaging with genetically encoded fluorescent markers to quantify lysosomal and mitochondrial dynamics in live cells. This will reveal how GBA responds to different α-syn dosages and how that affects cellular homeostasis. Second, I will leverage isogenic lines containing APOE allelic variants and GBA knockdown. GBA and APOE are involved in lipid metabolism, suggesting a putative link between lipid metabolism disruption and α-syn pathology, although there is no published connection between the two proteins. I will use next generation genomic sequencing and proteomic approaches to determine the interacting effects between SNCA, GBA, and APOE allelic variants on α-syn pathology severity. Completion of this aim will uncover whether GBA and APOE are acting through converging or independent pathways, expanding our knowledge of the network that contributes to disease pathology. By coupling multiple genetic risk factors into a multi-cell type model, I will pioneer new technology and approaches to dissect epistatic mechanisms that influence α-syn pathology. These insights will guide us in developing new therapeutic approaches, enabling earlier detection, treatment, and prevention of synucleinopathies.

项目摘要 突触核蛋白病，包括路易体痴呆（DLB）和帕金森病（PD），是一种日益增长的疾病。 健康危机，影响美国100多万人，全球估计有1000万人。 突触核蛋白病是由α-突触核蛋白（α-syn）在大脑中积聚成非天然纤维引起的 并且可以表现为运动症状、认知症状或两者的组合。目前有 没有有效的治疗方法来治愈或预防突触核蛋白病，在很大程度上是因为没有明确的原因， 疾病有许多确定的遗传和环境风险因素与突触核蛋白病， 但这些因素导致疾病的背景仍不清楚。除了直接突变SNCA之外， 编码α-syn的基因、GBA和APOE突变是公认的风险因素， 显示增加α-syn聚集。然而，这些风险因素中没有一个是疾病的重要预测因素， 这表明未知的遗传和环境相互作用可能会影响疾病的发生、严重程度和 α-syn病理学的临床结局。在这个建议中，我专注于细胞类型和基因型如何相互作用， 彼此调节α-syn病理严重性。我假设α-syn的病理是由上位性 破坏不同细胞类型内稳态的遗传因素之间的相互作用， 神经元易受有害的α-syn病理和神经变性的影响。我将使用以下方法来研究这个假设： 两个独立的，互补的方法来剖析细胞和分子机制，调节 α-syn表型。首先，我将GBA敲除与SNCA三倍化、野生型和敲除细胞系结合。 GBA突变和α-syn聚集之间存在明确的联系，但这些蛋白质如何连接仍然是未知的。 未知我将使用高内涵成像与基因编码的荧光标记，以量化溶酶体和 活细胞中的线粒体动力学。这将揭示GBA如何对不同的α-syn剂量做出反应， 影响细胞内稳态其次，我将利用含有APOE等位基因变体和GBA的等基因系， 击倒。GBA和APOE参与脂质代谢，提示脂质代谢与GBA和APOE之间可能存在联系。 代谢破坏和α-syn病理学，尽管这两种蛋白质之间没有公开的联系。 我将使用下一代基因组测序和蛋白质组学方法来确定相互作用的影响 SNCA、GBA和APOE等位基因变体对α-syn病理严重程度的影响。完成这一目标将揭示 GBA和APOE是否通过融合或独立的途径起作用，扩大了我们对 对疾病病理学有贡献的网络。通过将多种遗传风险因素耦合到多细胞类型中， 模型，我将开拓新的技术和方法来剖析影响α-syn的上位机制， 病理这些见解将指导我们开发新的治疗方法，使早期检测， 治疗和预防共核蛋白病。