Elucidating the role of LRRK2 in the gastrointestinal manifestation of PD

项目摘要 先前的研究结果表明炎症性肠病（IBD）和帕金森病之间存在生物学联系 （PD）。然而，可能将肠道炎症与PD进展联系起来的确切机制仍然未知。 有趣的是，PD和IBD都与LRRK 2基因突变有关;特别是，最近的遗传分析表明， 由项目3的PI Peter领导的研究将LRRK 2变异体N2081 D与PD和IBD的风险增加联系起来。然而， 目前尚不清楚LRRK 2N 2081 D变异是否在这些进行性疾病的发展中起作用。 因此，本项目的总体目标是研究LRRK 2在α-突触核蛋白诱导的病理学中的作用 并有可能扩散到大脑。该方法是基于评估诱导α- LRRK 2N 2081 D突变体及其野生型在淋巴细胞和回肠/结肠肠神经元中突触核蛋白过表达， 类型对应物。为了获得细胞特异性，我们将采用优化的（腺相关）病毒递送系统 其能够实现有效的系统性和器官特异性基因转导。我们将使用这些尖端的病毒 技术诱导淋巴细胞中的系统性α-突触核蛋白过表达和淋巴细胞中的局部过表达。 在LRRK 2N 2081 D突变体和野生型小鼠中的回肠/结肠肠神经元。我们将评估肠道动力， 缺陷、粪便生物标志物、肠道微生物组组成以及内源性小鼠α-突触核蛋白 聚合和扩散。因此，我们的具体目标是（1）评估LRRK 2N 2081 D变异体的影响 对全身淋巴细胞与局部回肠/结肠肠神经元pS129-α-syn聚集和病理学的影响;（2） 检查LRRK 2N 2081 D变体对粪便生物标志物和肠道微生物组组成的影响， 淋巴细胞和肠α-突触核蛋白过表达;（3）检查LRRK 2N 2081 D变体对 淋巴细胞和肠道α-突触核蛋白过度表达后的帕金森病行为和细胞表型。我们 预期肠溶后LRRK 2 N2081 D突变体脑中的运动恶化和α-突触核蛋白聚集更大。 α-突触核蛋白转染。另一方面，我们预期pS129-α-syn对淋巴细胞的侵袭减弱， α-突触核蛋白转染。这些研究的结果可能揭示了早期干预的新机会， PD进展的过程。

Project Summary Previous findings suggest a biological link between Inflammatory bowel disease (IBD) and Parkinson’s Disease (PD). However, the exact mechanism potentially linking gut inflammation to PD progression remains unknown. Interestingly, PD and IBD are both linked to mutations in the LRRK2 gene; specifically, recent genetic analyses led by Project 3’s PI Peter linked the LRRK2 variant N2081D to increased risk for both PD and IBD. It is however unknown whether the LRRK2N2081D variation plays a role in the development of these progressive disorders. Accordingly, the overall aim of this project is to examine the role of LRRK2 in α-synuclein-induced pathologies in gut tissue and their potential spread to brain. The approach is based on assessing the effects of inducing α- synuclein overexpression in lymphocytes and ileum/colon enteric neurons of LRRK2N2081D mutants and their wild- type counterparts. To attain cell specificity, we will employ optimized (adeno-associated) viral delivery systems that enable efficient systemic and organ-specific gene transduction. We will employ these cutting-edge viral techniques to induce systemic α-synuclein overexpression in lymphocytes and local overexpression in ileum/colon enteric neurons in LRRK2N2081D mutants and wild-type mice. We will assess gut motility, motor deficits, stool-based biomarkers, gut microbiome composition, as well as endogenous mouse α-synuclein aggregation and spreading. Accordingly, our Specific Aims are (1) to assess the impact of LRRK2N2081D variants on systemic lymphocytes vs. local ileum /colon enteric neuron pS129-α-syn aggregation and pathology; (2) to examine the impact of LRRK2N2081D variants on stool-based biomarkers and gut microbiome composition upon lymphocytic and enteric α-synuclein overexpression; (3) to examine the impact of LRRK2N2081D variants on Parkinsonian behavioral and cellular phenotypes upon lymphocytic and enteric α-synuclein overexpression. We expect greater motor deterioration and α-synuclein aggregation in the brain of LRRK2 N2081D mutants upon enteric α-synuclein transfection. On the other hand, we expect attenuated brain pS129-α-syn invasion upon lymphocytic α-synuclein transfection. The outcome of these studies may reveal novel opportunities for early interventions in the course of PD progression.