SUMMARY
# Epigenetic Regulation Dysfunction in Alzheimer's and Parkinson's Disease
## Background and Rationale
Neurodegenerative diseases like Alzheimer's (AD) and Parkinson's disease (PD) exhibit complex pathophysiology involving genetic, environmental, and epigenetic factors. Growing evidence suggests that epigenetic dysregulation plays a crucial role in disease onset and progression, affecting gene expression patterns critical for neuronal survival and function. This comprehensive clinical study aims
METHODOLOGY NOTES
Phase 1: Sample Collection and Processing (Months 1-6): Collect post-mortem brain tissue from frontal cortex, hippocampus, and substantia nigra regions from AD (n=60), PD (n=60), and control (n=40) subjects. Establish iPSC lines from patient fibroblasts using Sendai virus reprogramming. Phase 2: iPSC Differentiation (Months 4-12): Differentiate iPSCs into dopaminergic neurons using dual-SMAD inhibition protocol with SB431542 and LDN193189, followed by FGF8 and SHH treatment. Generate cholinergic neurons using NGF and BDNF supplementation. Validate neuronal identity using immunofluorescence for TH, ChAT, and MAP2. Phase 3: Epigenetic Profiling (Months 7-18): Extract high-quality DNA and RNA using AllPrep DNA/RNA Mini Kit. Perform RRBS library preparation using MspI digestion and bisulfite conversion. Conduct ChIP-seq using antibodies against H3K4me3, H3K27me3, and H3K9ac with 10μg chromatin per immunoprecipitation. Generate RNA-seq libraries including small RNA fractions using TruSeq pr