Validation of hub genes in apoE-/- atherosclerotic mice

Validation Score: 0.800 Price: $0.50 Atherosclerosis apoE-/- mice Status: proposed

What This Experiment Tests

Validation experiment designed to validate causal mechanisms targeting C1QA, C1QC, SPI1 in apoE-/- mice. Primary outcome: Validation of high expression levels of C1QA, C1QC, and SPI1

Description

This animal model experiment utilized apolipoprotein E knockout (apoE-/-) mice, a well-established model of atherosclerosis, to validate the expression of identified hub genes in vivo. These mice spontaneously develop atherosclerotic lesions similar to human disease when fed a normal or high-fat diet. The expression levels of C1QA, C1QC, and SPI1 were measured using quantitative real-time PCR in tissues from these atherosclerotic mice. This experiment provided crucial in vivo validation of the computational findings, demonstrating that the identified hub genes are indeed upregulated in a physiologically relevant animal model of atherosclerosis.

TARGET GENE
C1QA, C1QC, SPI1
MODEL SYSTEM
apoE-/- mice
ESTIMATED COST
$0
TIMELINE
0 months
PATHWAY
Complement signaling pathway
SOURCE
extracted_from_pmid_38179058
PRIMARY OUTCOME
Validation of high expression levels of C1QA, C1QC, and SPI1

Scoring Dimensions

Info Gain 0.00 (25%) Feasibility 0.00 (20%) Hyp Coverage 0.00 (20%) Cost Effect. 0.00 (15%) Novelty 0.00 (10%) Ethical Safety 0.00 (10%) 0.800 composite

📖 Wiki Pages

C1QC ProteinproteinC1QCgeneSPI1 Gene - PU.1geneC1QA Gene — Complement Component 1q A ChaingeneC1QA GenegeneAPOE — Apolipoprotein EgeneGWAS Findings Hub: Alzheimer's and Parkinson's DismechanismCBS Single-Cell Transcriptomics MechanismsmechanismGWAS Findings Hubmechanism

Protocol

Validation of Hub Genes in apoE-/- Atherosclerotic Mice Protocol

Phase 1: apoE-/- Mouse Breeding and Atherosclerosis Induction (Days 1-42)

Mouse Maintenance: Maintain apoE-/- mice (C57BL/6J-Apoe<tm1Unc>/J, JAX #002052) on standard chow or Western diet (WD, 21% milk fat, 0.2% cholesterol) for 8 or 16 weeks. Compare with age-matched C57BL/6J WT controls. Housing: 12h light/dark cycle, ad libitum food/water.

Dietary Groups: (a) Standard chow 8 weeks (n=12), (b) WD 8 weeks (n=15), (c) WD 16 weeks (n=15). Randomize mice into groups at weaning (3 weeks). Record body weight weekly.

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Expected Outcomes

Primary Outcomes

Hub Gene Upregulation: In WD-fed apoE-/- mice (16 weeks), expect C1QA upregulation ≥3.5-fold (p < 0.001), C1QC upregulation ≥2.8-fold (p < 0.001), and SPI1 upregulation ≥2.0-fold (p < 0.01) vs. WT chow controls. 4 additional hub genes show ≥1.8-fold change (p < 0.05).

Macrophage Correlation: C1QA immunofluorescence intensity correlates with aortic lesion area (Pearson's r = 0.72, p = 0.003) and CD68+ area (r = 0.81, p < 0.001), supporting macrophage source of complement proteins.

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Success Criteria

Primary Success Criteria

Statistical Significance: Top 3 hub genes (C1QA, C1QC, SPI1) must show ≥2.0-fold upregulation in WD apoE-/- vs. WT chow controls with p < 0.01 (Student's t-test, n≥4 biological replicates per group).

Biological Replication: Fold-change direction must be consistent across all replicates within each group (≥80% of replicates showing same direction of change as group mean).

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Related Hypotheses (5)

Complement C1q Mimetic Decoy Therapy0.695
Complement C1QA Spatial Gradient in Cortical Layers0.678
Complement C1q Subtype Switching0.665
Complement-Mediated Synaptic Pruning Dysregulation0.612
Complement-Mediated Synaptic Protection0.580

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