Gut Microbiome-Derived Metabolites in Alpha-Synuclein Propagation

SUMMARY

# Gut Microbiome-Derived Metabolites in Alpha-Synuclein Propagation ## Background and Rationale This translational clinical study investigates the role of gut microbiome-derived metabolites in alpha-synuclein aggregation and propagation, addressing a critical knowledge gap in Parkinson's disease (PD) pathogenesis. Growing evidence supports the gut-brain axis hypothesis in neurodegeneration, where altered intestinal microbiota may contribute to alpha-synuclein misfolding and subsequent neuronal d

METHODOLOGY NOTES

Phase 1 (Months 1-6): Recruit 120 PD patients (Hoehn-Yahr stages I-III) and 80 healthy controls from movement disorder clinics. Obtain informed consent and perform baseline clinical assessments including MDS-UPDRS, Montreal Cognitive Assessment, and constipation scoring. Collect fecal samples (10g) in sterile containers, flash-freeze at -80°C within 2 hours. Phase 2 (Months 7-12): Extract microbial DNA using QIAamp PowerFecal Pro DNA Kit, perform 16S rRNA V4 region sequencing on Illumina MiSeq platform. Conduct untargeted metabolomics using LC-MS/MS with HILIC and reverse-phase chromatography. Target analysis of 150+ metabolites including SCFAs, indoles, phenolics, and bile acids. Phase 3 (Months 13-20): Isolate PBMCs from 50ml blood samples using Ficoll density gradient centrifugation. Culture primary neurons differentiated from patient iPSCs (n=20 per group) for 21 days. Treat cells with identified metabolites at physiologically relevant concentrations (1-100μM) for 72 hours. Phase 4