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epiMuller README

Muller plot image

About
Author

Jennifer L. Havens

Purpose

Visualize lineages overtime, with phylogenetic context, based on viral genomes.

Language

Python3

Inputs

timetree, ancestral state reconstruction (Nextstain JSON file or annotated TreeTime nexus file), sample collection dates and, PANGO lineages (optional)

Workflow overview
  • epimuller-parse (optional): parse fasta names with 'bar isodate' suffix into usable fasta and metadata files.
  • epimuller: wrapper for epimuller-define and epimuller-draw.
    • epimuller-define: assigns samples to clades based on ancestral reconstruction of specified aa mutations or trait (hierarchy), and counts number of samples in a clade withen each time frame (abundance).
    • epimuller-draw: plots the frequency clades overtime, as specifed by abundance and hierarchy inputs from epimuller-define.
Source code avaliblity

gitHub

Documentation avaliblity

Read the Docs

Quick start

pip3 install epimuller

epimuller [-h] [-oDir OUTDIRECTORY] -oP OUTPREFIX
		(-n INNEXTSTRAIN | -a ANNOTATEDTREE) -m
		INMETA [-p INPANGOLIN] [--noPangolin]
		[-k TRAITOFINTERSTKEY]
		[-f TRAITOFINTERSTFILE] [-g GENEBOUNDRY]
		[-mut VOCLIST [VOCLIST ...]]
		[-t TIMEWINDOW]
		[-s STARTDATE] [-e ENDDATE] [-mt MINTIME]
		[-min MINTOTALCOUNT] [-c CASES_NAME]
		[--avgWindow AVGWINDOW]
		[-l {date,time,bimonthly}]
		[-lp {Right,Max,Start,End}] [--WIDTH WIDTH]
		[--HEIGHT HEIGHT] [--LEGENDWIDTH LEGENDWIDTH]
		[--MARGIN MARGIN] [--FONTSIZE FONTSIZE]
		[--LABELSHIFT LABELSHIFT]

SOME EXAMPLES

Examples for full run

To see steps used to prep files for these examples look at scripts/Example_CommandsFromScratch.txt on gitHub.

Visulize default aa mutation list
epimuller \
	-n inputData/GISAID_NYCPHL_04_29/02_nextstrainResults \
	-m inputData/GISAID_NYCPHL_04_29/gisaid_2021_04_30_00_rename.tsv \
	-oDir 03_results_NYCPHL_April29 \
	-oP 01_defaultAAList \
	-c inputData/CITY_US-NY_NYC_outbreakinfo_epidemiology_data_2021-04-30.tsv
Visulize a trait: lineage
epimuller \
	-n inputData/GISAID_NYCPHL_04_29/02_nextstrainResults \
	-m inputData/GISAID_NYCPHL_04_29/gisaid_2021_04_30_00_rename.tsv \
	-oDir 03_results_NYCPHL_April29 \
	-oP 02_pangolin \
	-c inputData/CITY_US-NY_NYC_outbreakinfo_epidemiology_data_2021-04-30.tsv \
	--traitOfInterstFile traits.json \
	--traitOfInterstKey lineage \
	-lp Max \
	-min 100 \
Visulize your own aa mutation list
epimuller \
	-n inputData/GISAID_NYCPHL_04_29/02_nextstrainResults \
	-m inputData/GISAID_NYCPHL_04_29/gisaid_2021_04_30_00_rename.tsv \
	-oDir 03_results_NYCPHL_April29 \
	-oP 03_selectedAA \
	-c inputData/CITY_US-NY_NYC_outbreakinfo_epidemiology_data_2021-04-30.tsv \
	-mut 'SE484K' 'S*452*' \
	-min 50 \ 
	-mt 20
Visulize default aa mutation list with TreeTime input
epimuller \
	-a inputData/GISAID_NYCPHL_04_29/06_treetimeDates_aa/timetree.nexus \
	-oDir 04_results_NYCPHL_April29 \
	-oP defaultAA_treetime \
	-m inputData/GISAID_NYCPHL_04_29/gisaid_2021_04_30_00_rename.tsv \
	-g data/geneAAboundries.json \
	--FONTSIZE 18
Visulize a trait: lineage with TreeTime input
epimuller \
	-a inputData/GISAID_NYCPHL_04_29/06_treetimeDates_aa/timetree.nexus \
	-oDir 03_results_NYCPHL_April29 \
	-oP 05_pangolin_treetime \
	-m inputData/GISAID_NYCPHL_04_29/gisaid_2021_04_30_00_rename.tsv \
	--traitOfInterstKey lineage \
	--noPangolin #does not label with mode of pangolin lineages in clade, label clade with defining lineage only 

Known edge cases / featrues to add

Known edge cases which are not correctly dealt with or features I intend to address (eventually). If you run into anything else please let me know with an issue on gitHub.

	- feel free to ignore the undefined.svg that gets made - it is related to checking the size of the text to space out labels

	- allow combination of aa mutants, not just 1
	- define polytomy behavior
	- option for user defined col names in metadata
	- auto detect

Addtional features

Color

If you would like to specify color for clade: in --parentHierarchy_name file (of epimuller-draw/drawMuller.py input) add col with name: "color" and hex color value (starting with #) for clades you want to specify.

Parse GISAID fasta for metadata

epimuller-parse If you have downloaded sequences from GISAID under the search tab, you can parse out the names into a metadata file (format tested as of 2021-04-30).

epimuller arguments

epimuller [-h] [-oDir OUTDIRECTORY] -oP OUTPREFIX
		(-n INNEXTSTRAIN | -a ANNOTATEDTREE) -m
		INMETA [-p INPANGOLIN] [--noPangolin]
		[-k TRAITOFINTERSTKEY]
		[-f TRAITOFINTERSTFILE] [-g GENEBOUNDRY]
		[-mut VOCLIST [VOCLIST ...]]
		[-t TIMEWINDOW]
		[-s STARTDATE] [-e ENDDATE] [-mt MINTIME]
		[-min MINTOTALCOUNT] [-c CASES_NAME]
		[--avgWindow AVGWINDOW]
		[-l {date,time,bimonthly}]
		[-lp {Right,Max,Start,End}] [--WIDTH WIDTH]
		[--HEIGHT HEIGHT] [--LEGENDWIDTH LEGENDWIDTH]
		[--MARGIN MARGIN] [--FONTSIZE FONTSIZE]
		[--LABELSHIFT LABELSHIFT]

arguments:
  -h, --help						show this help message and exit
  -n INNEXTSTRAIN, --inNextstrain INNEXTSTRAIN
		nextstrain results with tree.nwk and
		[traitOfInterst].json (default: None)
  -a ANNOTATEDTREE, --annotatedTree ANNOTATEDTREE
		nexus file name with annotation:
		[&!traitOfInterst=value], as output by treetime
		(default: None)

Options for full repot:
  -oDir OUTDIRECTORY, --outDirectory OUTDIRECTORY
		folder for output (default: ./)
  -oP OUTPREFIX, --outPrefix OUTPREFIX
		prefix of out files withen outDirectory (default:
		None)

Options passed to epimuller-define:
  -m INMETA, --inMeta INMETA
		metadata tsv with 'strain' and 'date'cols, optional:
		cols of trait of interst; and pangolin col
		named:'pangolin_lineage', 'lineage' or 'pangolin_lin'
		(default: None)
  -p INPANGOLIN, --inPangolin INPANGOLIN
		pangolin output lineage_report.csv file, if argument
		not supplied looks in inMeta for col with
		'pangolin_lineage', 'pangolin_lin', or 'lineage'
		(default: metadata)
  --noPangolin				  do not add lineage to clade names (default: False)
  -k TRAITOFINTERSTKEY, --traitOfInterstKey TRAITOFINTERSTKEY
		key for trait of interst in json file OR (if
		-a/--annotatedTree AND key is mutations with aa (not
		nuc): use 'aa_muts') (default: aa_muts)
  -f TRAITOFINTERSTFILE, --traitOfInterstFile TRAITOFINTERSTFILE
		[use with -n/--inNextstrain] name of
		[traitOfInterstFile].json in '-n/--inNextstrain'
		folder (default: aa_muts.json)
  -g GENEBOUNDRY, --geneBoundry GENEBOUNDRY
		[use with -a/--annotatedTree AND -k/--traitOfInterst
		aa_muts] json formated file specifing start end
		postions of genes in alignment for annotatedTree (see
		example data/geneAAboundries.json) (default: None)
  -mut VOCLIST [VOCLIST ...], --VOClist VOCLIST [VOCLIST ...]
		list of aa of interest in form
		[GENE][*ORAncAA][site][*ORtoAA] ex. S*501*, gaps
		represed by X, wild card aa represented by * (default:
		None)
  -t TIMEWINDOW, --timeWindow TIMEWINDOW
		number of days for sampling window (default: 7)
  -s STARTDATE, --startDate STARTDATE
		start date in iso format YYYY-MM-DD or 'firstDate'
		which sets start date to first date in metadata
		(default: 2020-03-01)
  -e ENDDATE, --endDate ENDDATE
		end date in iso format YYYY-MM-DD or 'lastDate' which
		sets end date as last date in metadata (default:
		lastDate)

Options passed to epimuller-draw:
  -mt MINTIME, --MINTIME MINTIME
		minimum time point to start plotting (default: 30)
  -min MINTOTALCOUNT, --MINTOTALCOUNT MINTOTALCOUNT
		minimum total count for group to be included (default:
		50)
  -c CASES_NAME, --cases_name CASES_NAME
		file with cases - formated with 'date' in ISO format
		and 'confirmed_rolling' cases, in tsv format (default:
		None)
  --avgWindow AVGWINDOW
		width of rolling mean window in terms of
		--timeWindow's (recomend using with small
		--timeWindow) ; default: sum of counts withen
		timeWindow (ie no average) (default: None)
  -l {date,time,bimonthly}, --xlabel {date,time,bimonthly}
		Format of x axis label: ISO date format or timepoints
		from start, or dd-Mon-YYYY on 1st and 15th (default:
		date)
  -lp {Right,Max,Start,End}, --labelPosition {Right,Max,Start,End}
		choose position of clade labels (default: Right)

Options passed to epimuller-draw for page setup:
  --WIDTH WIDTH				 WIDTH of page (px) (default: 1500)
  --HEIGHT HEIGHT		   HEIGHT of page (px) (default: 1000)
  --LEGENDWIDTH LEGENDWIDTH
		LEGENDWIDTH to the right of plotting area (px)
		(default: 220)
  --MARGIN MARGIN		   MARGIN around all sides of plotting area (px)
		(default: 60)
  --FONTSIZE FONTSIZE
  --LABELSHIFT LABELSHIFT
		nudge label over by LABELSHIFT (px) (default: 15)

epimuller-define: make abundance and hiearchy files

epimuller-define  [-h] [-oDir OUTDIRECTORY] -oP OUTPREFIX
				   (-n INNEXTSTRAIN | -a ANNOTATEDTREE) -m INMETA
				   [-p INPANGOLIN] [--noPangolin]
				   [-k TRAITOFINTERSTKEY] [-f TRAITOFINTERSTFILE]
				   [-g GENEBOUNDRY] [-mut VOCLIST [VOCLIST ...]]
				   [-t TIMEWINDOW]
				   [-s STARTDATE] [-e ENDDATE]

optional arguments:
  -h, --help						show this help message and exit
  -oDir OUTDIRECTORY, --outDirectory OUTDIRECTORY
		folder for output (default: ./)
  -oP OUTPREFIX, --outPrefix OUTPREFIX
		prefix of out files withen outDirectory (default:
		None)
  -n INNEXTSTRAIN, --inNextstrain INNEXTSTRAIN
		nextstrain results with tree.nwk and
		[traitOfInterstFile].json (default: None)
  -a ANNOTATEDTREE, --annotatedTree ANNOTATEDTREE
		nexus file name with annotation:
		[&!traitOfInterstKey=value], as output by treetime
		(default: None)
  -m INMETA, --inMeta INMETA
		metadata tsv with 'strain' and 'date'cols, optional:
		col for [traitOfInterstKey]; and pangolin col named:
		'pangolin_lineage' 'lineage' or 'pangolin_lin'
		(default: None)
  -p INPANGOLIN, --inPangolin INPANGOLIN
		pangolin output lineage_report.csv file, if argument
		not supplied looks in inMeta for col with
		'pangolin_lineage', 'pangolin_lin', or 'lineage'
		(default: metadata)
  --noPangolin				  do not add lineage to clade names (default: False)
  -k TRAITOFINTERSTKEY, --traitOfInterstKey TRAITOFINTERSTKEY
		key for trait of interst in json file OR (if
		-a/--annotatedTree AND key is mutations with aa (not
		nuc): use 'aa_muts') (default: aa_muts)
  -f TRAITOFINTERSTFILE, --traitOfInterstFile TRAITOFINTERSTFILE
		[use with -n/--inNextstrain] name of
		[traitOfInterstFile].json in '-n/--inNextstrain'
		folder (default: aa_muts.json)
  -g GENEBOUNDRY, --geneBoundry GENEBOUNDRY
		[use with -a/--annotatedTree AND -k/--traitOfInterst
		aa_muts] json formated file specifing start end
		postions of genes in alignment for annotatedTree (see
		example data/geneAAboundries.json) (default: None)
  -mut VOCLIST [VOCLIST ...], --VOClist VOCLIST [VOCLIST ...]
		list of aa of interest in form
		[GENE][*ORAncAA][site][*ORtoAA] ex. S*501*, gaps
		represented by X, wild card aa represented by *
		(default: None)
  -t TIMEWINDOW, --timeWindow TIMEWINDOW
		number of days for sampling window (default: 7)
  -s STARTDATE, --startDate STARTDATE
		start date in iso format YYYY-MM-DD or 'firstDate'
		which is in metadata (default: 2020-03-01)
  -e ENDDATE, --endDate ENDDATE
		end date in iso format YYYY-MM-DD or 'lastDate' which
		is in metadata (default: lastDate)

epimuller-draw: plot

epimuller-draw  [-h] -p PARENTHIERARCHY_NAME -a ABUNDANCE_NAME
		[-c CASES_NAME] [--avgWindow AVGWINDOW] -o OUTFOLDER
		[-mt MINTIME] [-min MINTOTALCOUNT]
		[-l {date,time,bimonthly}] [-lp {Right,Max,Start,End}]
		[--WIDTH WIDTH] [--HEIGHT HEIGHT]
		[--LEGENDWIDTH LEGENDWIDTH] [--LABELSHIFT LABELSHIFT]
		[--MARGIN MARGIN] [--FONTSIZE FONTSIZE]

arguments:
  -h, --help						show this help message and exit
  -p PARENTHIERARCHY_NAME, --parentHierarchy_name PARENTHIERARCHY_NAME
		csv output from mutationLinages_report.py with child
		parent col (default: None)
  -a ABUNDANCE_NAME, --abundance_name ABUNDANCE_NAME
		csv output from mutationLinages_report.py with
		abundances of clades (default: None)
  -c CASES_NAME, --cases_name CASES_NAME
		file with cases - formated with 'date' in ISO format
		and 'confirmed_rolling' cases, in tsv format (default:
		None)
  --avgWindow AVGWINDOW
		width of rolling mean window in terms of
		--timeWindow's (recomend using with small
		--timeWindow) ; default: sum of counts withen
		timeWindow (ie no average) (default: None)
  -o OUTFOLDER, --outFolder OUTFOLDER
		csv output from mutationLinages_report.py with child
		parent col (default: None)
  -mt MINTIME, --MINTIME MINTIME
		minimum time point to start plotting (default: 30)
  -min MINTOTALCOUNT, --MINTOTALCOUNT MINTOTALCOUNT
		minimum total count for group to be included (default:
		50)
  -l {date,time,bimonthly}, --xlabel {date,time,bimonthly}
		Format of x axis label: ISO date format or timepoints
		from start, or dd-Mon-YYYY on 1st and 15th (default:
		date)
  -lp {Right,Max,Start,End}, --labelPosition {Right,Max,Start,End}
		choose position of clade labels (default: Right)

Options for page setup:
  --WIDTH WIDTH				 WIDTH of page (px) (default: 1500)
  --HEIGHT HEIGHT		   HEIGHT of page (px) (default: 1000)
  --LEGENDWIDTH LEGENDWIDTH
		LEGENDWIDTH to the right of plotting area (px)
		(default: 220)
  --LABELSHIFT LABELSHIFT
		nudge label over by LABELSHIFT (px) (default: 15)
  --MARGIN MARGIN		   MARGIN around all sides of plotting area (px)
		(default: 60)
  --FONTSIZE FONTSIZE

Install methods

With Bioconda
conda install -c bioconda epimuller
With pip
pip3 install epimuller

#If there is an issue with cairo, try:

pip3 install pycairo
pip3 install epimuller
From source

Download source code from gitHub or pypi

#open as needed for download format
tar -zxvf epimuller-[version].tar.gz

cd epimuller-[version]

python3 setup.py install
Run scripts directly

Note you will have to install all dependencies.

Download source code from gitHub or pypi

#open as needed for download format
tar -zxvf epimuller-[version].tar.gz

cd epimuller-[version]

#to run epimuller
python3 ./scripts/mutationLinages_report.py [arugments]

#to run epimuller-parse
python3 ./scripts/parseFastaNames.py  [arugments]

#to run epimuller-define 
python3 ./scripts/defineAndCountClades.py  [arugments]

#to run epimuller-draw 
python3 ./scripts/drawMuller.py  [arugments]

Citation

Please link to this github if you have used epimuller in your research.

Extra notes on GISAID

If you do use GISAID data please acknowledge the contributers, such as with language suggested by GISAID.

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Visualize lineages overtime, with phylogenetic context, based on viral genomes

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