Joining different datasets and analyzing using the group-specific condition effects strategy (DESeq2)
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@ac76911c
Last seen 2.8 years ago
United States

Hello,

I would like to summon Michael Love here and anyone else that can contribute. I was doing some research on DESeq2 and reading the following topic of the tutorial: Group-specific condition effects, individuals nested within groups, I was wondering if I can use this strategy to join different datasets, then each dataset will be treated as a group, each group will have their distinct individuals, and each individual will have two conditions (in common among all datasets). Does the design described account for the probable batch effect?

Thank you in advance.

DESeq2 • 774 views
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@mikelove
Last seen 3 days ago
United States

Yes, this sounds reasonable and will estimate a fixed batch/dataset effect and help you estimate a fixed condition effect. If you wanted to get more complicated, you could attempt to estimate a random effect of condition, but this isn't supported within DESeq2. For what it's worth, I know there are some methods for RNA-seq that account for sample non-independence with random effects but I don't know of methods where the random effects are not nuisance parameters, but parameters of interest.

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Michael Love Could I please ask if a similar method would work my set of samples? I have 255 samples which have been generated from 76 subjects (46 with a disease and 30 controls). The samples were collected from these subjects at two timepoints. And these samples were then treated or untreated. My main question was to see the differences between disease and controls when treated at either timepoint (less interested at differences between timepoints).

Initially, was going to use a simpler design ~ grouped factor (Treated/Untreated x Disease/Control x Timepoint A/B) but wondered if I could account, in the model, for differences between subjects that are not attributable to treatment, disease or timepoint. The design ~ subject + group failed I think because subject can only have a disease or be a control. ~60% subjects have samples across timepoints and almost always (>90%) have paired treated and untreated samples.

Could you please advise what would work within DESeq 2?

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There is an example of how you can do this in the DESeq vignette.

An alternative would be to fit a linear mixed model using the variancePartition package, which will not require complete cases.

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