To avoid confusion with the diffHic R package (https://bioconductor.org/packages/release/bioc/html/diffHic.html), HiCdiff has been renamed to HiCcompare (https://github.com/dozmorovlab/HiCcompare).
HiCdiff provides functions for joint normalization and difference detection in multiple Hi-C datasets. HiCdiff operates on processed Hi-C data in the form of chromosome-specific chromatin interaction matrices. HiCdiff is available as an R package, the major releases can be found on Bioconductor (here)[insert URL].
HiCdiff accepts three-column tab-separated text files storing chromatin interaction matrices in a sparse matrix format which are available from several sources such as the http://aidenlab.org/data.html and http://cooler.readthedocs.io/en/latest/index.html. HiCdiff is designed to give the user the ability to perform a comparative analysis on the 3-Dimensional structure of the genomes of cells in different biological states. HiCdiff first can jointly normalize two Hi-C datasets to remove biases between them. Then it can detect signficant differences between the datsets using a genomic distance based permutation test. The novel concept of the MD plot, based on the commonly used MA plot or Bland-Altman plot is the basis for these methods. The log Minus is plotted on the y axis while the genomic Distance is plotted on the x axis. The MD plot allows for visualization of the differences between the Hi-C datasets.
The main functions are:
hic_loess()which performs jointloessnormalization on the Hi-C datasetshic_diff()which performs the difference detection process to detect significant changes between Hi-C datasets and assist in comparative analysis
Several Hi-C datasets are also included in the package.
First make sure you have all dependencies installed in R.
install.packages(c('dplyr', 'data.table', 'ggplot2', 'gridExtra',
'mgcv', 'parallel', 'devtools'))
source("https://bioconductor.org/biocLite.R")
biocLite("InteractionSet")
To install HiCdiff from bioconductor open R and enter the following commands.
# Currently in submission process; for now use github version
## try http:// if https:// URLs are not supported
# source("https://bioconductor.org/biocLite.R")
# biocLite("HiCdiff")
# library(HiCdiff)
Or to install HiCdiff directly from the github release open R and enter the following commands.
library(devtools)
install_github('dozmorovlab/HiCdiff', build_vignettes = TRUE)
library(HiCdiff)
First you will need to obtain some Hi-C data. Data is available from the sources listed in the overview along with many others. You will need to extract the data and read it into R as either a 3 column sparse upper triangular matrix or a 7 column BEDPE file. For more details on data extraction see the vignette included with HiCdiff.
Below is an example analysis using HiCdiff. The data in 3 column sparse upper triangular matrix format is loaded and the first step is to create a hic.table object using the create.hic.table() function. Next, the two Hi-C matrices are jointly normalized using the hic_loess() function. Finally, difference detection can be performed using the hic_diff() function. The hic_loess() and hic_diff() functions will also produce an MD plot for visualizing the differences between the datasets.
# load data
library(HiCdiff)
data("HMEC.chr22")
data("NHEK.chr22")
# create the `hic.table` object
chr22.table = create.hic.table(HMEC.chr22, NHEK.chr22, chr = 'chr22')
head(chr22.table)
# Jointly normalize data for a single chromosome
hic.table = hic_loess(chr22.table, Plot = TRUE)
head(hic.table)
# input hic.table object into hic_diff
hic.table = hic_diff(hic.table, Plot = TRUE)
head(hic.table)
Refer to the HiCdiff vignette for full usage instructions. For a full explanation of the methods used in HiCdiff see the manuscript here.
To view the usage vignette:
browseVignettes("HiCdiff")
The HiCdiff paper included several supplemental files that showcase some of the usage and reasoning behind the methods. Below are the titles and brief descriptions of each of these vignettes along with links to the compiled .pdf and the source .Rmd files.
Normalization method comparison.
Comparison of several Hi-C normalization techniques to display the persistence of bias in individually normalized chromatin interaction matrices, and its effect on the detection of differential chromatin interactions.
S2 File. Estimation of the IF power-law depencence.
Estimation of the power-law depencence between the
S3 File. Estimation of the SD power-law dependence.
Estimation of the power-law depencence between the
S4 File. Estimation of proportion of zeros.
Estimation of the depencence between the proportion of zeros and distance between interacting regions. This vignette shows distribution of zeros in real Hi-C data. The results were used for modeling the proportion of zeros in simulated Hi-C matrices with a linear function.
S5 File. Evaluation of difference detection in simulated data.
Extended evaluation of differential chromatin interaction detection analysis using simulated Hi-C data. Many different classifier performance measures are presented. Note: if trying to compile the source .Rmd this will take a long time to knit.
S6 File. Evaluation of difference detection in real data.
Extended evaluation of differential chromatin interaction detection analysis using real Hi-C data. Many different classifier performance measures are presented. Note: if trying to compile the source .Rmd this will take a long time to knit.
S7 File. loess at varying resolution.
Visualization of the loess loint normalization over varying resolutions. This vignette shows that increasing sparsity of Hi-C matrices with increasing resolution causes loess to become less useful for normalization at high resolutions.
Please cite HiCdiff if you use it in your analysis.
HiCdiff: A method for joint normalization of Hi-C datasets and differential chromatin interaction detection John Stansfield, Mikhail G. Dozmorov bioRxiv 147850; doi: https://doi.org/10.1101/147850
Suggestions for new features and bug reports are welcome. Please create a new issue for any of these or contact the author directly: @jstansfield0 ([email protected])
Authors: @jstansfield0 ([email protected]) & @mdozmorov ([email protected])