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SigProfilerMatrixGenerator

SigProfilerMatrixGenerator creates mutational matrices for all types of somatic mutations. It allows downsizing the generated mutations only to parts for the genome (e.g., exome or a custom BED file). The tool seamlessly integrates with other SigProfiler tools.

INTRODUCTION

The purpose of this document is to provide a guide for using the SigProfilerMatrixGenerator framework to generate mutational matrices for a set of samples with associated mutational catalogues. An extensive Wiki page detailing the usage of this tool can be found at https://osf.io/s93d5/wiki/home/.

For users that prefer working in an R environment, a wrapper package is provided and can be found and installed from: https://github.com/AlexandrovLab/SigProfilerMatrixGeneratorR

PREREQUISITES

The framework is written in PYTHON, however, it also requires the following software with the given versions (or newer):

  • PYTHON version 3.4 or newer
  • WGET version 1.9 or RSYNC if you have a firewall

By default the installation process will save the FASTA files for all chromosomes for the default genome assemblies (GRCh37, GRCH38, mm10, mm9, rn6). As a result, ~3 Gb of storage must be available for the downloads for each genome.

QUICK START GUIDE

This section will guide you through the minimum steps required to create mutational matrices:

  1. Install the python package using pip:
                          pip install SigProfilerMatrixGenerator
  1. a. Install your desired reference genome from the command line/terminal as follows (a complete list of supported genomes can be found below): 
    $ python
>> from SigProfilerMatrixGenerator import install as genInstall
>> genInstall.install('GRCh37', rsync=False, bash=True)

     This will install the human 37 assembly as a reference genome. You may install as many genomes as you wish. If you have a firewall on your server, you may need to install rsync and use the rsync=True parameter. Similarly, if you do not have bash, 
 use bash=False.
    b. To install a reference genome that you have saved locally, you can do the following: 
    $ python
>> from SigProfilerMatrixGenerator import install as genInstall
>> genInstall.install('GRCh37', offline_files_path='path/to/directory/containing/GRCh37.tar.gz')
  2. Place your vcf files in your desired output folder. It is recommended that you name this folder based on your project's name
  3. From within a python session, you can now generate the matrices as follows:
$ python3
>>from SigProfilerMatrixGenerator.scripts import SigProfilerMatrixGeneratorFunc as matGen
>>matrices = matGen.SigProfilerMatrixGeneratorFunc("test", "GRCh37", "/Users/ebergstr/Desktop/test",plot=True, exome=False, bed_file=None, chrom_based=False, tsb_stat=False, seqInfo=False, cushion=100)

The layout of the required parameters are as follows:

  SigProfilerMatrixGeneratorFunc(project, reference_genome, path_to_input_files)

where project, reference_genome, and path_to_input_files must be strings (surrounded by quotation marks, ex: "test"). Optional parameters include:

  exome=False:       [boolean] Downsamples mutational matrices to the exome regions of the genome
  bed_file=None      [string path to bed_file] Downsamples mutational matrices to custom regions of the genome. Requires the full path to the BED file. 
  chrom_based=False  [boolean] Outputs chromosome-based matrices
  plot=False         [boolean] Integrates with SigProfilerPlotting to output all available visualizations for each matrix. 
  tsb_stat=False     [boolean] Outputs the results of a transcriptional strand bias test for the respective matrices. 
  seqInfo=True      [boolean] Ouputs original mutations into a text file that contains the SigProfilerMatrixGenerator classificaiton for each mutation. 
  cushion=100	[integer] Adds an Xbp cushion to the exome/bed_file ranges for downsampling the mutations.

INPUT FILE FORMAT

This tool currently supports maf, vcf, simple text file, and ICGC formats. The user must provide variant data adhering to one of these four formats. If the user’s files are in vcf format, each sample must be saved as a separate files.

Output File Structure

The output structure is divided into three folders: input, output, and logs. The input folder contains copies of the user-provided input files. The outputfolder contains a DBS, SBS, ID, and TSB folder (there will also be a plots folder if this parameter is chosen). The matrices are saved into the appropriate folders. The logs folder contains the error and log files for the submitted job.

COPY NUMBER MATRIX GENERATION

In order to generate a copy number matrix, provide the an absolute path to a multi-sample segmentation file obtained from one of the following copy number calling tools (if you have individual sample files, please combine them into one file with the first column corresponding to the sample name):

  1. ASCAT
  2. ASCAT_NGS
  3. SEQUENZA
  4. ABSOLUTE

In addition, provide the name of the project and the output directory for the resulting matrix. The final matrix will be placed in a folder with the name of the project in the directory specified by the output path.

An example to generate the CNV matrix is as follows:

$ python3

>>from SigProfilerMatrixGenerator.scripts import CNVMatrixGenerator as scna
>>file_type = "ASCAT_NGS"
>>input_file = "./SigProfilerMatrixGenerator/references/CNV/all.breast.ascat.summary.sample.tsv" #example input file for testing
>>output_path = "./SigProfilerMatrixGenerator/references/CNV/"
>>project = "Breast_Cancer"
>>scna.generateCNVMatrix(file_type, input_file, project, output_path)

SUPPORTED GENOMES

This tool currently supports the following genomes:

GRCh38.p12 [GRCh38] (Genome Reference Consortium Human Reference 38), INSDC Assembly GCA_000001405.27, Dec 2013. Released July 2014. Last updated January 2018. This genome was downloaded from ENSEMBL database version 93.38.

GRCh37.p13 [GRCh37] (Genome Reference Consortium Human Reference 37), INSDC Assembly GCA_000001405.14, Feb 2009. Released April 2011. Last updated September 2013. This genome was downloaded from ENSEMBL database version 93.37.

GRCm39 [mm39] (Genome Reference Consortium Mouse Reference 39), INSDC Assembly GCA_000001635.9, Jun 2020. Last updated August 2020. This genome was downloaded from ENSEMBL database version 103.

GRCm38.p6 [mm10] (Genome Reference Consortium Mouse Reference 38), INDSDC Assembly GCA_000001635.8, Jan 2012. Released July 2012. Last updated March 2018. This genome was downloaded from ENSEMBL database version 93.38.

GRCm37 [mm9] (Release 67, NCBIM37), INDSDC Assembly GCA_000001635.18. Released Jan 2011. Last updated March 2012. This genome was downloaded from ENSEMBL database version release 67.

Rnor_6.0 [rn6] INSDC Assembly GCA_000001895.4, Jul 2014. Released Jun 2015. Last updated Jan 2017. This genome was downloaded from ENSEMBL database version 96.6.

Epstein-Barr Virus [EBV] NC_007605.1, Nov 2005. Last updated Aug 2018. This genome was downloaded from the NCBI database: https://www.ncbi.nlm.nih.gov/nuccore/82503188/.

CanFam3.1 [dog] GCA_000002285.2, Sep 2011. Last updated Jun 2019. This genome was downloaded from ENSEMBL database version 100.

WBcel235 [c_elegans] GCA_000002985.3, Oct 2014. Last updated Jan 2019. This genome was downloaded from ENSEMBL database version 100.

*One can specify "_havana" to the end of the genome to include annotations in t-cell receptor genes and IG clusters (available for GRCh37, GRCh38, and mm10).

LOG FILES

All errors and progress checkpoints are saved into sigProfilerMatrixGenerator_[project]_[genome].err and sigProfilerMatrixGenerator_[project]_[genome].out, respectively. For all errors, please email the error and progress log files to the primary contact under CONTACT INFORMATION.

CITATION

Bergstrom EN, Huang MN, Mahto U, Barnes M, Stratton MR, Rozen SG, and Alexandrov LB (2019) SigProfilerMatrixGenerator: a tool for visualizing and exploring patterns of small mutational events. BMC Genomics 20, Article number: 685. https://bmcgenomics.biomedcentral.com/articles/10.1186/s12864-019-6041-2

COPYRIGHT

Copyright (c) 2019, Erik Bergstrom [Alexandrov Lab] All rights reserved.

Redistribution and use in source and binary forms, with or without modification, are permitted provided that the following conditions are met:

Redistributions of source code must retain the above copyright notice, this list of conditions and the following disclaimer.

Redistributions in binary form must reproduce the above copyright notice, this list of conditions and the following disclaimer in the documentation and/or other materials provided with the distribution.

THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS" AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL THE COPYRIGHT HOLDER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED OF THE POSSIBILITY OF SUCH DAMAGE.

CONTACT INFORMATION

Please address any queries or bug reports to Erik Bergstrom at [email protected]. Please address any queries or bug reports related to CNV's or SV's to Azhar Khandekar at [email protected]. Additional support can be provided by Mark Barnes at [email protected].