Hello,
I have been told I cannot use WGCNA with only two conditions (control and treatment). I am not sure why that would be the case and I would like to confirm if this is true? I have 3 samples for each condition so a total of 6 samples and I am not correlating to a quantifiable trait, I am only correlating the modules to the conditions.
Thank you
2 answers
I believe it is not the condition but the number of replicates that are required for running WGCNA. Typically, you are required to have more than 15 samples. Ideally, I would recommend more than 30 replicates for each condition.
Here is the list of frequently asked questions: https://horvath.genetics.ucla.edu/html/CoexpressionNetwork/Rpackages/WGCNA/faq.html
Another advice would be plotting PCA/MDS plots to check if there are no Batch Effects or technical effects such as sex, non-biologicals effects.
Good luck!
I posted this as an answer outside of Biostars:
There is no correct or incorrect answer to this. To me, it makes more sense to perform WGCNA separately, so, one network for disease and one network for control:
Analysis 1:
- generate network in Control
- identify modules and hub genes
- perform gene enrichment on the genes in each module
Analysis 2:
- generate network in Disease
- identify modules and hub genes
- perform gene enrichment on the genes in each module
Analysis 3:
- discuss differences between hub genes between Disease network and Control network
- discuss differences between modules (and their gene enrichments) between Disease network and Control network
If you generate a single network for all samples (Disease + Control), then, after you identify modules, you can still try this: perform a linear regression between DiseaseStatus and the module eigenvalues:
summary(lm(BlueModule ~ DiseaseStatus))
summary(lm(RedModule ~ DiseaseStatus))
summary(lm(YellowModule ~ DiseaseStatus))
*et cetera*
Then, in this way, you would focus on the modules from which the regression produces p<0.05, as these modules may contain information about DiseaseStatus.
Kevin
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