Host - Microbe interactions in Parkinson's using a Caenorhabditis elegans (C. elegans)- Escherichia coli (E. coli) Holobiont Model

帕金森病（PD）是一种进行性神经系统疾病，其表现为对神经系统运动系统的逐渐退行性影响。这种临床表现在病理上源于a- Synuclein （aS）的异常积累和错误折叠，在路易小体中形成神经毒性的不溶性淀粉样蛋白原纤维。这会导致大脑黑质中产生多巴胺的神经元的退化。导致仍然无法治愈和第二普遍的神经退行性疾病的发展，PD在65岁以上的老年人中更为普遍。最近的研究提供了证据，表明中枢神经系统的外周信号，特别是来自胃肠道和肠道微生物群的信号，参与了PD的进展。例如，aS病理开始于外周组织，如肠道，并逐渐扩散到大脑的多个区域。人的胃肠道包含了体内最密集的微生物共生生态系统。总的来说，这些主要由细菌组成的微生物产生的蛋白质编码基因比人类基因组多100多倍。近年来，越来越多的证据表明，潜伏在胃肠道中的微生物对宿主的发育和健康至关重要。因为它们有助于维生素的生物合成，通过各种代谢途径改善能量和营养的吸收，并通过帮助免疫系统的发育和成熟来减轻肠道病原体感染，为资源提供竞争环境。肠道微生物群也被证明通过产生进入血液的代谢物来影响大脑功能。然而，肠道细菌如何影响aS聚集尚不清楚。为了研究微生物群对PD的影响，我们将使用秀丽隐杆线虫（C. elegans）作为模型来证明as聚集。秀丽隐杆线虫是一个理想的模型，因为它的肠道微生物群可以精确控制，在单物种和单基因水平上研究生理过程，秀丽隐杆线虫已被证实是研究帕金森病分子机制和蛋白质聚集的有价值的模型。利用一种成熟的秀丽隐杆线虫PD模型，在肌肉细胞中表达人类aS。表型分析将通过应用大肠杆菌KEIO敲除收集所有非必需基因和表型微阵列技术（Biolog）来分别揭示细菌遗传损伤和营养补充对aS蠕虫生理的影响。然后，为了进一步揭示这些相互作用在线虫aS聚集背景下的机制，RNA测序，代谢组学，光漂白后荧光恢复（FRAP），原子力显微镜（AFM）和许多其他技术将被用于aS聚集的生物学和生物物理特征。最终，在aS的背景下，宿主-微生物的相互作用可以帮助阐明PD相关的蛋白质聚集过程。

PhD project strategic theme: Bioscience for an integrated understanding of healthParkinson's disease (PD) is a progressive neurological disorder that manifest itself through its gradual degenerative effects on the motor system of the nervous system. This clinical manifestation is pathologically derived from the abnormal accumulation and misfolding of a- Synuclein (aS), forming neurotoxic insoluble amyloid fibrils in Lewy bodies. This leads to the degradation of dopamine producing neurons in the substantia nigra of the brain. Resulting in the development of the still incurable and second most prevalent neurodegenerative disease, PD which is more prevalent amongst the ageing population, of above 65 years of age.Recent research provides evidence that signals peripheral to the central nervous system, particularly from the gastrointestinal tract and gut microbiota, are involved in the progression of PD. For example, aS pathology begins in peripheral tissues such as in the intestines and gradually spreads to multiple brain regions.The human gastrointestinal tract encompasses the densest microbial symbiotic ecosystem within the body. Altogether, these microbes which primarily constitute of bacteria produces over a 100 times more protein-coding genes than the human genome. Recently, cumulative evidence suggests that microbes harbored in the gastrointestinal tract are fundamental to the development and health of their host. For they aid in vitamin biosynthesis, improved energy and nutrient uptake through various metabolic pathways, and alleviate intestinal pathogen infections by helping develop and maturate the immune system providing a competitive environment for resources. The gut microbiota has also been shown to impact brain function by producing metabolites that enter the bloodstream. However, how gut bacteria affect aS aggregation is still unclear.To investigate the effect of microbiota in PD, we will use, Caenorhabditis elegans (C. elegans) as a model to demonstrate aS aggregation. C. elegans is an ideal model because its gut microbiota can be precisely controlled to study physiological processes at both the single-species and single-gene levels, and C. elegans has been validated as a valuable model of studying molecular mechanisms of Parkinson's disease and protein aggregation.Using a well-established C. elegans model of PD that expresses human aS in muscle cells. Phenotypic assays will be employed through the application of the E. coli KEIO Knockout Collection for all nonessential genes and the Phenomic Microarrays technology (Biolog), to respectively reveal the effects of bacterial genetic impairment and nutritional supplementation on the physiology of aS worms. Then, to further unravel the mechanisms underlying these interactions in the context of aS aggregation in the worms, RNA Sequencing, metabolomics, fluorescence recovery after photobleaching (FRAP), atomic force microscopy (AFM) and many more techniques will be employed for biological and biophysical characterisation of aS aggregation.Ultimately, host - microbe interactions in the context of aS could help to elucidate the processes involved in the protein aggregation associated in PD.