11 Jun 2022
Coming from a quantitative genetics background, correcting for multiple comparisons meant controlling the family-wise error rate (FWER) using a procedure like Bonferroni correction. This all changed when I took John Storey’s “Advanced Statistics for Biology” class in grad school. John is an expert in statistical interpretation of high-dimensional data and literally wrote the book, well paper, on false-discovery rate (FDR) as an author of Storey & Tibshirani 2006. His description of the FDR has grounded my interpretation of hundreds of genomic datasets and I’ve continued to pay this knowledge forward with dozens of white-board style descriptions of the FDR for colleagues. As an interviewer and paper reviewer I still regularly see accomplished individuals and groups where “FDR control” is a clear blind spot. In this post I’ll layout how I whiteboard the FDR problem, and then highlight a specialized application of the FDR for “denoising” genomic datasets.
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08 May 2022
Timecourses are a powerful experimental design for evaluating the impact of a perturbation. These perturbations are usually chemicals because chemicals, such as a drug, can be introduced quickly and with high temporal precision. Although, with some technologies, such as the estradiol-driven promoters that I used in the induction dynamics expression atlas (IDEA), it is possible to rapidly perturb a single gene further increasing specificity and broadening applicability. By rapidly perturbing individuals, they can be synchronized based on the time when dosing began. We often call this point when dosing begins “time zero” while all subsequent measurements correspond to the time post perturbation. (Since time zero corresponds to a point when a perturbation is applied, but will not yet impact the system, this measurement is usually taken before adding the perturbation.)
One of the benefits of collecting a time zero measurement is it allows us to remove, or account for, effects that are shared among all time points. In many cases this may just amount to analyzing fold-changes of post-perturbation measurement with respect to their time zero observation, rather than the original measurements themselves. This can be useful if there is considerable variation among timecourses irrespective of the perturbation, such as if we were studying humans or mice. Similarly, undesirable variation due to day-to-day variation in instruments, sample stability, or any of the many other factors which could produce batch effects, can sometimes by addressed by measuring each timecourse together and working with fold-changes. In either case, correcting for individual effects using pre-perturbation measurements will increase our power to detect perturbations’ effects.
Aside from correcting for unwanted variation, the kinetics of timecourses are a rich source of information which can be either a blessing or a curse. With temporal information, ephemeral responses can be observed. We can see both which features are changing and when they are changing. And, the ordering of events can point us towards causality. In practice, each of these goals can be difficult, or impossible to achieve, leaving us with a nagging feeling that we’re leaving information on the table. There are many competing options for identifying differences in timecourses, few ways of summarizing dynamics in an intuitive way, and causal inference is often out of reach. In this post, and others to follow, I’ll pick apart a few of these limitations, discussing developments that were applied to the IDEA, but will likely be useful for others thinking about biological timeseries analysis (or other timeseries if you are so inclined!). Here, I evaluate a few established methods for identifying features which vary across time and then introduce an alternative approach based on the Multivariate Gaussian distribution and Mahalanobis distance which increases power and does not require any assumptions about responses’ kinetics.
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08 May 2022
Romic is an R package, which I developed, that is now is now available on CRAN. There is already a nice README for romic on GitHub and a pkgdown site, so here, I will add some context regarding the problems this package addresses.
The first problem we’ll consider is that genomics data analysis involves a lot of shuffling between various forms of wide and tall data and incrementally tacking on attributes as needed. Romic aims to simplify this process, by providing a set of flexible data structures that accommodate a range of measurements and metadata and can be readily inter-converted based on the needs of an analysis.
The second challenge we’ll contend with is decreasing the time it takes to generate a plot so that mechanics of plotting rarely interrupt the thought process of data interpretation. Building upon romic’s data structure, the meaning of variables (feature-, sample-, measurement-level) are encoded in a schema, so they can be appropriately surfaced to filter or reorder a dataset, and add ggplot2 aesthetics. Interactivity is facilitated using Shiny apps composed from romic-centric Shiny modules.
Both of these solutions increase the speed, clarity, and succinctness of analysis. I’ve developed and will continue to refine this package to save myself (and hopefully others!) time.
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