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regionReport: Interactive reports for region-based analyses

[version 1; peer review: 1 approved, 1 approved with reservations, 1 not approved]
PUBLISHED 01 May 2015
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Abstract

regionReport is an R package for generating detailed interactive reports from regions of the genome. The report includes quality-control checks, an overview of the results, an interactive table of the genomic regions and reproducibility information. regionReport can easily be expanded with report templates for other specialized analyses. In particular, regionReport has an extensive report template for exploring derfinder results from annotation-agnostic RNA-seq differential expression analyses.

Keywords

Report, Interactive, Reproducibility, Genomics, Sequencing, ChIP-seq, RNA-seq, Software

Introduction

Many analyses of genomic data result in regions along the genome that associate with a covariate of interest. These genomic regions can result from identifying differentially bound peaks from ChIP-seq data1, identifying differentially methylated regions (DMRs) from DNA methylation data2, or performing base-resolution differential expression analyses using RNA sequencing data3,4. The genomic regions themselves are commonly stored in a GRanges object from GenomicRanges5 when working with R or the BED file format on the UCSC Genome Browser6, but other information on these regions, for example summary statistics on the magnitude of effects and statistical significance, also provide useful information. The usage of R in genomics is increasingly common due to the usefulness and popularity of the Bioconductor project7, and in the latest version (3.0), 181 unique packages use GenomicRanges for many workflows, demonstrating the widespread utility of identifying and summarizing characteristics of genomic regions.

Here we introduce regionReport which allows users to explore genomic regions of interest through interactive stand-alone HTML reports that can be shared with collaborators. These reports are flexible enough to display plots and quality control checks within a given experiment, but can easily be expanded to include custom visualizations and conclusions. The resulting HTML report emphasizes reproducibility of analyses8 by including all the R code without obstructing the resulting plots and tables. We envision regionReport will provide a useful tool for exploring and sharing genomic region-based results from high throughput genomics experiments.

Methods

Implementation

The package includes a R Markdown template which is processed using knitr9 and rmarkdown10, then styled using knitrBootstrap11. This package generates a HTML report that includes a series of plots for checking the quality of the results and browsing the table of regions. Each element of the report has a brief explanation, although actual interpretation of the results is dataset- and workflow-dependent. To facilitate navigation a menu is always included, which is useful for users interested in a particular section of the report. Figure 1, panel a shows the menu of the general report for a set of regions with associated p-values. The code for each plot or table is hidden by default and can be shown by clicking on the appropriate toggle as shown in Figure 2.

a4285c5d-3dda-4e00-8db2-3c01e41e1ed4_figure1.gif

Figure 1. regionReport workflow.

Example region input, the appropriate regionReport function to use, and menu of the resulting report for the general use case (panel a) and derfinder results (panel b).

a4285c5d-3dda-4e00-8db2-3c01e41e1ed4_figure2.gif

Figure 2. Interactively display the code for each table/figure in the report.

View by default (panel a) and after clicking on the “R source” toggle (panel b) for a section of the general report. The full report is available at the supplementary website and includes a toggle to hide/show all the R code.

Quality checks

This section of the report includes a variety of quality control steps which help the user determine whether the results are sensible. The quality control steps explore:

  • P-values, Q-values, and FWER adjusted p-values

  • Region width

  • Region area: sum of single-base level statistics (if available)

  • Mean coverage or other score variables (if available)

A combination of density plots and numerical summaries are used in these quality checks. If there are statistically significant regions, the distributions are compared between all regions and the significant ones. For example, the distribution region widths might have a high density of small values for the global results, but shifted towards higher values for the subset of significant regions.

Genomic overview

The report includes plots to visualize the location of all the regions as well as the significant ones. Differences between them can reveal location biases. The nearest known annotation feature for each region is summarized and visually inspected in the report.

Best regions

An interactive table with the top 500 (default) regions is included in this section. This allows the user to sort the region information according to their preferred ranking option. For example, lowest p-value, longest width, chromosome, nearest annotation feature, etc. The table also allows the user to search and subset it interactively. A common use case is when the user wants to check if any of the regions are near a known gene of their interest.

Reproducibility

At the end of the report, detailed information is provided on how the analysis was performed. This includes the actual function call to generate the report, the path where the report was generated, time spent, and the detailed R session information including package versions of all the dependencies.

The R code for generating the plots and tables in the report is included in the report itself, thus allowing users to manually reproduce any section of the report, customize them, or simply change the graphical parameters to their liking.

derfinder report

When exploring derfinder results, for each of the best 100 (default) DERs a plot showing the coverage per sample is included in the report. These plots allow the user to visualize the differences identified by derfinder along known exons, introns and isoforms. The plots are created using derfinderPlot, also available via Bioconductor.

Due to the intrinsic variability in RNA-seq coverage data or mapping artifacts, in situations where there are two candidate DERs that are relatively close there might be reasons to consider them a single candidate DER and its important to visualize them. This tailored report groups candidate DERs into clusters based on a distance cutoff. After ranking them by their area, for the top 20 (default) clusters it plots tracks with the coverage by sample, the mean coverage by group, the identified candidate DERs colored by whether they are statistically significant, and known alternative transcripts. Figure 1, panel b shows the main categories of the report generated from a richer region data set than in the general case.

Operation

Installation. regionReport and required dependencies can be easily installed from Bioconductor with the following commands:

source(“http://bioconductor.org/biocLite.R”)

biocLite(“regionReport”)

Input. To generate the report, the user first has to identify the regions of interest according to their analysis workflow. For example, by performing bumphunting to identify DMRs with bumphunter. The report is then created using renderReport() which is the main function in this package as shown in Figure 1, panel a. The argument customCode can be used to customize the report if necessary.

For the derfinder use case, the derfinderReport() function creates the recommended report that includes visualizations of the coverage information for the best regions and clusters of regions.

Output. A small example can be generated using:

example(“renderReport”, “regionReport”, ask=FALSE)

The resulting HTML file will open in the users default browser. Note that alternative output formats such as PDF files can also be generated, although they are not as dynamic and interactive as the HTML format.

Use cases

The supplementary website contains reports using DiffBind, bumphunter and derfinder results. The derfinder use case is illustrated with data sets previously described in 3 which span simulation results, a moderately sized data set (25 samples), and a large data set with 487 samples; thus covering a wide range of scenarios.

Summary

regionReport creates interactive reports from a set of regions and can be used in a wide range of genomic analyses. Reports generated with regionReport can easily be extended to include further quality checks and interpretation of the results specific to the data set under study. These shareable documents are very powerful when exploring different parameter values of an analysis workflow or applying the same method to a wide variety of data sets. The reports allow users to visually check the quality of the results, explore the properties of the genomic regions under study, and inspect the best regions and interactively explore them.

Furthermore, regionReport promotes reproducibility of data exploration and analysis. Each report provides R code that can be used as the starting point for other analyses within a dataset. regionReport provides a flexible output for exploring and sharing results from high throughput genomics experiments.

Software availability

Software access

regionReport is freely available via Bioconductor at bioconductor.org.

http://leekgroup.github.io/regionReportSupp/ hosts the code for generating three types of reports as well as the resulting HTML reports generated by regionReport. Versions of all software used are included in the reports.

Latest source code

The latest source code is available at Bioconductor and github.com/leekgroup/regionReport via the git-svn bridge although we recommend users to install regionReport directly from Bioconductor.

Archived source code as at the time of publication

Archived source code available at http://dx.doi.org/10.5281/zenodo.17083

License

Artistic-2.0.

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CITE
how to cite this article
Collado-Torres L, Jaffe AE and Leek JT. regionReport: Interactive reports for region-based analyses [version 1; peer review: 1 approved, 1 approved with reservations, 1 not approved]. F1000Research 2015, 4:105 (https://doi.org/10.12688/f1000research.6379.1)
NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.
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Open Peer Review

Current Reviewer Status: ?
Key to Reviewer Statuses VIEW
ApprovedThe paper is scientifically sound in its current form and only minor, if any, improvements are suggested
Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
Not approvedFundamental flaws in the paper seriously undermine the findings and conclusions
Version 1
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PUBLISHED 01 May 2015
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Reviewer Report 22 Jun 2015
David Robinson, Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA 
Approved
VIEWS 0
The authors present regionReport, an R package to produce interactive HTML reports from a genomic-region based analysis, such as those produced by derfinder, bumphunter or DiffBind. The report shows quality control summaries, interactive tables of the most significant regions, and ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Robinson D. Reviewer Report For: regionReport: Interactive reports for region-based analyses [version 1; peer review: 1 approved, 1 approved with reservations, 1 not approved]. F1000Research 2015, 4:105 (https://doi.org/10.5256/f1000research.6840.r8559)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
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Reviewer Report 11 Jun 2015
Karthik Ram, Berkeley Institute for Data Science, University of California Berkeley, Berkeley, USA 
Approved with Reservations
VIEWS 0
Review of regionReport: Interactive reports for region based analysis.

This short software tool article describes a new R package, `regionReport`, available from Bioconductor that generates HTML reports which allows users to explore genomic regions and quickly scan quality control information. The ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Ram K. Reviewer Report For: regionReport: Interactive reports for region-based analyses [version 1; peer review: 1 approved, 1 approved with reservations, 1 not approved]. F1000Research 2015, 4:105 (https://doi.org/10.5256/f1000research.6840.r8558)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
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Reviewer Report 18 May 2015
Timothy J. Triche Jr, Jane Anne Nohl Division of Hematology, USC/Norris Comprehensive Cancer Center, Keck School of Medicine of the University of Southern California, Los Angeles, CA, USA 
Not Approved
VIEWS 0
Needs more figures to demonstrate why a user would choose this tool.  For example http://leekgroup.github.io/regionReportSupp/bumphunter-example/index.html (but even better would be to show an example, e.g. ITGB2 exon inclusion/exclusion or multiscale DMRs, where in our hands at least, nothing else short of ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Triche Jr TJ. Reviewer Report For: regionReport: Interactive reports for region-based analyses [version 1; peer review: 1 approved, 1 approved with reservations, 1 not approved]. F1000Research 2015, 4:105 (https://doi.org/10.5256/f1000research.6840.r8554)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
  • Author Response 18 May 2015
    Jeffrey Leek, Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, 21205, USA
    18 May 2015
    Author Response
    Thanks, we will update with more figures and expand the description. This is meant to be a short description of the software but we certainly appreciate the feedback on how ... Continue reading
COMMENTS ON THIS REPORT
  • Author Response 18 May 2015
    Jeffrey Leek, Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, 21205, USA
    18 May 2015
    Author Response
    Thanks, we will update with more figures and expand the description. This is meant to be a short description of the software but we certainly appreciate the feedback on how ... Continue reading

Comments on this article Comments (0)

Version 2
VERSION 2 PUBLISHED 01 May 2015
Comment
Alongside their report, reviewers assign a status to the article:
Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested
Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions
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