🧫 Experiment Protocol
In VivoAlzheimer's diseaseTREM25xFAD transgenic mice (n=15/group x 4 groups: WT-vehicle, 5xFAD-vehicle, 5xFAD-AL002c low, 5xFAD-AL002c high)proposed
Test whether a TREM2-agonist antibody (AL002c analogue) normalises the dysfunctional astrocyte-microglia cross-talk that amplifies neuroinflammation in 5xFAD mice. Weekly IV injections (10 mg/kg) from 3 to 9 months of age; endpoint readouts at 9 months include LPS-stimulated astrocyte A1-marker expression (C3, Lcn2), microglial plaque-associated gene scores (Cst7, Spp1), soluble Abeta42/Abeta40 ratio, and NOR cognitive test.
PRIMARY OUTCOME
Reduction of astrocyte A1 markers (C3, Lcn2 mRNA) >=40% vs 5xFAD-vehicle; NOR discrimination index >=0.65
EXPECTED OUTCOMES
AL002c-analogue will suppress astrocyte A1 conversion by >=40% (C3, Lcn2), shift microglia toward DAM-protective state (increased Cst7, Spp1), reduce plaque burden by ~25%, and restore NOR discrimination index to >=0.65.
SUCCESS CRITERIA
Primary: >=40% reduction in C3 mRNA in GFAP+ astrocytes (Student t-test p<0.05). Secondary: NOR discrimination index >=0.65 (vs ~0.50 vehicle); >=15% plaque area reduction.
PROTOCOL
1. Breed 5xFAD hemizygous x C57BL/6J; genotype at P21. 2. Randomise to 4 groups at 3 months (n=15/group, sex-balanced). 3. Administer AL002c-analogue or vehicle IV weekly (3-9 months). 4. Monthly behavioural: NOR, open field. 5. Sacrifice at 9 months; isolate cortical microglia (CD11b+) and astrocytes (GFAP+) by FACS. 6. Bulk RNA-seq on sorted populations; ELISA for Abeta42/40; IHC plaque burden. 7. Primary endpoint: fold-change in A1 marker panel (C3, Lcn2, H2-D1); secondary: plaque load, NOR score, microglial DAM signature.
Source: task:a989715e-c687-4558-91ca-74fce1474bd2