Is it recommended to analyze different tissues together when using edgeR
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Lars Roed • 0
@1db20970
Last seen 27 days ago
Denmark

I am analyzing an experiment where I have samples from multiple regions of the brain, in a number of different conditions. We are only interested in differences within brain regions, not between them.

Is it recommended to combine the tissues into a single model, or should they be analyzed separately?

My initial thought was that increasing the number of samples should provide a better estimate of the mean-variance relationship, and increase the power to detect differences. When comparing the two approaches some regions seem to gain power to detect differences, while other lose power when combined into a single model.

DifferentialExpression edgeR • 540 views
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@gordon-smyth
Last seen 2 hours ago
WEHI, Melbourne, Australia

It is a matter of judgement. If the regions are very similar in expression and the samples have been collected and processed at the same time as part of the same study, then you might benefit from analyzing them all together. If the regions are quite different in expression, or differ in variability, then it is better to analyze them separately.

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Thank you, I'm guessing I could plot the average logCPMs of the tissues against each other to determine if the expression differs, is it possible to use e.g. plotBCV or plotQLDisp to determine if the tissues differ in variability?

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Just plot the samples with plotMDS().

Alternatively you would use voomLmFit() with sample.weights=TRUE or var.group=tissue. In principle, you could even use voomByGroup() with group=tissue (https://github.com/YOU-k/voomByGroup). Either approach would equalize any systematic differencees in variability between tissue. I'm not necessarily recommending these approaches, just listing them as possiblities.

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