INTERACTIONS BETWEEN LYSOSOMAL STORAGE DISORDER GENES, SPHINGOLIPID HOMEOSTASIS, AND ALPHA-SYNUCLEIN MECHANISMS IN PARKINSON'S DISEASE

项目摘要 帕金森病(PD)是一种无法治愈的神经退行性疾病，有强有力的证据表明 遗传力。葡萄糖脑苷酶(GBA)变异杂合子的存在可增加帕金森病 与非携带者相比，风险增加了五倍。然而，部分损失的机制 葡萄糖脑苷酶功能对帕金森病易感性的影响尚不清楚。完全丧失 葡萄糖脑苷酶功能导致54例罕见常染色体隐性遗传病之一的高谢病 或被称为溶酶体储存障碍(LSD)的X连锁疾病。最近在我的 实验室表明，LSD基因变异的负担与帕金森病的风险显著相关，甚至 排除了GBA，这表明除GBA外，LSD基因可能对 帕金森病的发病机制。有趣的是，几个相关的基因在一个共享的 鞘磷脂代谢途径，类似于GBA。在我的项目中，我将使用果蝇 研究LSD基因功能部分或单倍性缺失破坏的假说 鞘磷脂代谢，导致溶酶体应激增强，增加了对 与PD相关的应激源，如α-突触核蛋白毒性和衰老。我的项目利用了我的 初步数据，我对300多个转基因果蝇品系进行了筛选，鉴定出21个LSD 基因敲除增强了α-突触核蛋白的毒性。我将确认这一机制 增强以确定这些LSD基因的部分丢失可能如何导致帕金森病 敏感度。我的项目还通过使用不同的方法解决了部分基因丢失在帕金森病中的作用 GBA功能缺失突变体的优势。我会分析这些突变体的剂量依赖性 使用基于质谱学的脂类组学干扰鞘磷脂代谢，之前 试图进一步干扰鞘磷脂代谢与其他LSD基因一起修改 神经退行性变。我们预计我们的结果将显著提高对 LSD基因缺失与帕金森病易感性的相互作用，包括潜在的剂量依赖关系 当前模型中未解决的交互作用。

Project Summary Parkinson’s disease (PD) is an incurable neurodegenerative disorder with strong evidence for heritability. The heterozygous presence of variants in glucocerebrosidase (GBA) increases PD risk by five-fold compared to non-carriers. However, the mechanism by which the partial loss of glucocerebrosidase function contributes to PD susceptibility is unknown. Complete loss of glucocerebrosidase function results in Gaucher’s disease, one of 54 rare autosomal recessive or X-linked diseases known as lysosomal storage disorders (LSDs). Recently work from my laboratory showed that burden of LSD gene variants significantly associated with PD risk, even to the exclusion of GBA, indicating that LSD genes in addition to GBA may contribute to the onset of PD pathogenesis. Interestingly, several of the implicated genes function within a shared sphingolipid metabolism pathway, similar to GBA. For my project, I will use Drosophila to investigate the hypothesis that partial or haploinsufficient loss of LSD gene function disrupts sphingolipid metabolism, leading to enhanced lysosomal stress and increased vulnerability to PD-related stressors such as α-synuclein toxicity and aging. My project take advantage of my preliminary data, in which I screened over 300 transgenic Drosophila lines to identify 21 LSD genes whose knockdown enhanced α-synuclein toxicity. I will confirm the mechanism of enhancement to determine how partial loss of these LSD genes might contribute to PD susceptibility. My project also addresses the role of partial gene loss in PD by using different strengths of GBA loss of function mutants. I will analyze these mutants for the dose-dependent disruption of sphingolipid metabolism using mass spectrometry-based lipidomics, before attempting to further perturb sphingolipid metabolism with other LSD genes to modify neurodegeneration. We expect our results to significantly improve understanding of the interaction between LSD gene loss and PD susceptibility, including potential dose-dependent interactions that are not addressed in current models.