📋 Research Plan
alzheimerAPOE4$2,400,00018 monthsN = 180
Assay
Protocol: AAV9-mediated prime editor delivery targeting astrocyte-specific GFAP promoter.
Reagents: PE3max prime editor (SpCas9-H840A-UNG), APOLLO targeting spacer, 3' NLS, 120nt ssODT.
Dose: 1×10^11 vg/mouse (low), 5×10^11 (mid), 2.5×10^12 (high), via intracerebroventricular injection.
Controls: AAV9-EGFP treated age-matched mice, APOE3 homozygous mice.
Readouts: (1) amyloid plaque burden (Thioflavin S, 6E10 IHC), (2) cognitive assessment (Morris water maze, Y-maze), (3) lipid transport assay (HDL-C quantification), (4) neuroinflammation markers (Iba1+ microglia count).
Timeline: 18 months total (3 month pre-op + 12 month treatment + 3 month assessment).
Primary endpoint
Change in amyloid plaque burden (%) from baseline at 12 months post-treatment
Kill criteria
- Plaque burden reduction < 20% at any dose tier
- Off-target edits > 5% in any tissue
- Grade 3+ adverse events in > 15% of treated animals
- No detectable APOE4→APOE3 conversion in target tissue
Power: {'test': 'two-sample t-test', 'alpha': 0.05, 'power': 0.8, 'effect_size': 0.45, 'allocation_ratio': 1.0, 'expected_dropout': 0.1}