In-vivo Studies in 5xFAD Mouse Model
Objective: Determine whether inhibition of pro-inflammatory gene expression and promotion of amyloid clearance is feasible in neuroinflammation and neurodegeneration mouse models, specifically using 5xFAD mice
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Software1
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Protocol Steps
Weighted Co-expression Network Analysis (WGCNA)
Apply WGCNA to existing microglial transcriptomic datasets from neuroinflammatory and neurodegenerative disease mouse models to identify modules of highly co-expressed genes
Note: Analysis performed on existing datasets
View evidence from paper
“Weighted co-expression network analysis (WGCNA) was applied to existing microglial transcriptomic datasets from neuroinflammatory and neurodegenerative disease mouse models to identify modules of highly co-expressed genes”
Module Contrast Analysis
Contrast identified modules with known signatures of homeostatic microglia and DAM (disease-associated microglia) to reveal novel molecular heterogeneity within DAM
Note: Comparative analysis of gene expression patterns
View evidence from paper
“These modules were contrasted with known signatures of homeostatic microglia and DAM to reveal novel molecular heterogeneity within DAM”
Flow Cytometric Validation
Perform flow cytometric validation studies to confirm existence of distinct DAM sub-populations in AD mouse models predicted by WGCNA
Note: Validation performed in AD mouse models
View evidence from paper
“Flow cytometric validation studies were performed to confirm existence of distinct DAM sub-populations in AD mouse models predicted by WGCNA”
Gene Ontology and Bioinformatics Analysis
Perform gene ontology analyses coupled with bioinformatics approaches to reveal drug targets and transcriptional regulators of microglial modules predicted to favorably modulate neuroinflammation in AD
Note: Computational analysis to identify therapeutic targets
View evidence from paper
“Gene ontology analyses coupled with bioinformatics approaches revealed drug targets and transcriptional regulators of microglial modules predicted to favorably modulate neuroinflammation in AD”
In-vivo Studies in 5xFAD Mouse Model
Conduct in-vivo studies in 5xFAD mouse model of neuroinflammation and neurodegeneration to determine whether inhibition of pro-inflammatory gene expression and promotion of amyloid clearance is feasible
Note: Studies guided by results from gene ontology and bioinformatics analyses
View evidence from paper
“These guided in-vivo and in-vitro studies in mouse models of neuroinflammation and neurodegeneration (5xFAD) to determine whether inhibition of pro-inflammatory gene expression and promotion of amyloid clearance was feasible”
In-vitro Studies
Conduct in-vitro studies in mouse models of neuroinflammation and neurodegeneration to complement in-vivo findings
Note: Parallel studies to in-vivo work
View evidence from paper
“These guided in-vivo and in-vitro studies in mouse models of neuroinflammation and neurodegeneration (5xFAD)”
Human Relevance Integration
Determine human relevance of findings by integrating results with AD genome-wide association studies and human AD and non-disease post-mortem brain proteomes
Note: Translational validation step
View evidence from paper
“We determined the human relevance of these findings by integrating our results with AD genome-wide association studies and human AD and non-disease post-mortem brain proteomes”