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 突触核酸症，包括痴呆症患有路易的痴呆症（DLB）和帕金森氏病（PD），是一个增长 健康危机，影响美国超过100万人，估计全球1000万人。 突触核苷是由蛋白质α-突触核蛋白（α-Syn）积累到大脑中非天然原纤维中引起的 并且可以表现出运动症状，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苷，在很大程度上是因为没有明确的理由 疾病。有许多与突触核苷有关的遗传和环境危险因素， 但是，这些因素导致疾病的背景仍然未知。除了直接对SNCA突变， 编码α-syn的基因，对GBA和APOE的突变是良好的危险因素，已经是 显示以增加α-syn聚集。但是，这些危险因素都不是疾病的重要预测因素 他们自己的，表明未知的遗传和环境相互作用可能会影响主动性，严重性和 α-Syn病理学的临床结果。在此提案中，我关注细胞类型和基因型如何与 彼此调节α-Syn病理严重程度。我假设α-syn病理是由认识的 破坏不同细胞类型中稳态的遗传因素之间的相互作用，并留下多巴胺能 神经元容易受到有害α-Syn病理学和神经退行性的影响。我将使用 两种独立的，完整的方法来剖析调节的细胞和分子机制 α-syn表型。首先，我将GBA击倒与SNCA- triplication，野生型和淘汰细胞系。 GBA突变与α-syn聚集之间存在明确的联系，但是这些蛋白如何连接仍然 未知。我将使用具有一般编码的荧光标记的高内感成像来量化溶酶体和 活细胞中的线粒体动力学。这将揭示GBA如何应对不同的α-Syn剂量以及如何反应 影响细胞稳态。其次，我将利用包含APOE等位基因变体和GBA的同源线 击倒。 GBA和APOE参与了脂质代谢，这表明脂质之间存在假定的联系 代谢破坏和α-Syn病理学，尽管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 SNCA，GBA和APOE等位基因在α-Syn病理严重程度上。这个目标的完成将揭开 GBA和APOE是通过融合还是独立的途径行事，扩大了我们对 有助于疾病病理的网络。通过将多个遗传危险因素耦合到多细胞类型 模型，我将开拓新技术和方法，以剖析影响α-Syn的认识机制 病理。这些见解将指导我们开发新的治疗方法，从而实现早期检测， 治疗和预防突触核心病。