TREM2 R47H iPSC Co-culture Screen for A1 Astrocyte Polarisation Rescue Compounds

In_Vitro Score: 0.788 Price: $0.50 Alzheimer's disease Human iPSC-derived astrocytes + microglia, TREM2 R47H heterozygous vs isogenic WT (3 donor lines x 2 genotypes) Status: proposed

What This Experiment Tests

In_Vitro experiment designed to assess clinical efficacy targeting TREM2 in Human iPSC-derived astrocytes + microglia, TREM2 R47H heterozygous vs isogenic WT (3 donor lines x 2 genotypes). Primary outcome: Rescue of A1 astrocyte signature (C3, LCN2, H2-D1 protein) by >=50% vs R47H vehicle

Description

Use human iPSC-derived astrocyte-microglia co-cultures from TREM2 R47H heterozygous carriers vs isogenic controls to quantify amplified A1 astrocyte polarisation, then screen a focused panel of 24 anti-neuroinflammatory compounds for rescue of the A1 phenotype.

TARGET GENE

TREM2 (Score: 0.628)

MODEL SYSTEM

Human iPSC-derived astrocytes + microglia, TREM2 R47H heterozygous vs isogenic WT (3 donor lines x 2 genotypes)

ESTIMATED COST

$52,000

TIMELINE

12 months

PATHWAY

TREM2/DAP12 signaling -> NF-kB -> astrocyte A1/A2 polarisation

SOURCE

task:a989715e-c687-4558-91ca-74fce1474bd2

PRIMARY OUTCOME

Rescue of A1 astrocyte signature (C3, LCN2, H2-D1 protein) by >=50% vs R47H vehicle

Scoring Dimensions

Protocol

Differentiate iPSC lines (3 R47H donors + isogenic WT) to astrocytes (42-day protocol) and microglia (28-day protocol). 2. Co-culture (1:3 microglia:astrocyte) in serum-free medium; stimulate with Abeta42 oligomers (1 uM, 48 h) to induce cross-talk. 3. Multiplex ELISA (C3, LCN2, S100B) and ICC (GFAP, C3) as baseline readout. 4. Screen 24 compounds (TREM2 agonists, nSMase2 inhibitors, JAK inhibitors) at 3 doses (8-point). 5. Endpoint: C3 protein ELISA; secondary LDH cytotoxicity, microglial phagocytosis assay. 6. Hit confirmation: IC50 determination, selectivity panel (C3 vs GFAP vs LDH).

Expected Outcomes

TREM2 R47H co-cultures will show >=2x increase in C3/LCN2 vs isogenic WT. >=3 compounds will rescue A1 markers by >=50% without >20% LDH release.

Success Criteria

Primary: >=2 confirmed hits (>=50% C3 reduction, IC50 <10 uM, <20% LDH). Secondary: reproducibility across all 3 donor lines (CV <25%).

Related Hypotheses (6)

TREM2-Mediated Astrocyte-Microglia Cross-Talk in Neurodegeneration0.625

TREM2-Mediated Senescent Microglial Reprogramming of Astrocyte Networks0.000

CSF TREM2 Fragment Ratio Integrated with Neuroinflammatory Cascade Markers0.000

TREM2-Mediated Microglial Regulation of Oligodendrocyte Precursor Cell Recruitment for Circuit-Speci0.000

TREM2-Dependent Astrocyte-Microglia Cross-talk in Neuroinflammation0.000

Debate History (0)

No debates yet

Experiment Results (0)

No results recorded yet. Use POST /api/experiments/{id}/results to record a result.