def doOntologyAnalysis(gene_lists, options):
'''do ontology analysis - requires
options.filename_assignments to be set.'''
E.info("reading association of categories and genes from %s" %
(options.filename_assignments))
gene2gos, go2infos = GO.ReadGene2GOFromFile(
IOTools.openFile(options.filename_assignments))
E.info("read %i ontologies" % (len(gene2gos)))
#############################################################
# sort out which ontologies to test
ontologies = options.ontology
# test all if none specified
if not ontologies:
ontologies = gene2gos.keys()
all_results = {}
for ontology in ontologies:
gene2go, go2info = gene2gos[ontology], go2infos[ontology]
if len(go2info) == 0:
E.warn("could not find information for terms - "
"could be mismatch between ontologies")
ngenes, ncategories, nmaps = GO.CountGO(gene2go)
E.info("%s: ontology assignments: %i genes mapped "
"to %i categories (%i maps)" %
(ontology, ngenes, ncategories, nmaps))
for geneset, x in gene_lists.iteritems():
foreground, background = x
E.debug("working on %s - %s" % (ontology, geneset))
E.info("%s - %s: (unfiltered) foreground=%i, "
"background=%i" %
(ontology, geneset, len(foreground), len(background)))
results = GO.AnalyseGO(gene2go, foreground, background)
if len(results.mSampleGenes) == 0:
E.warn("%s - %s: no genes with GO categories - "
"analysis aborted" % (ontology, geneset))
continue
# add sampling at this point for empirical FDR
all_results[(ontology, geneset)] = results
if options.fdr:
E.info("computing the FDR with method %s" % options.qvalue_method)
computeFDR(all_results, qvalue_method=options.qvalue_method)
outputOntologyResults(all_results, go2infos, options)
def main(argv=None):
parser = E.OptionParser(version="%prog version: $Id$",
usage=globals()["__doc__"])
parser.add_option("-s",
"--species",
dest="species",
type="string",
help="species to use [default=%default].")
parser.add_option("-i",
"--slims",
dest="filename_slims",
type="string",
help="filename with GO SLIM categories "
"[default=%default].")
parser.add_option("-g",
"--genes-tsv-file",
dest="filename_genes",
type="string",
help="filename with genes to analyse "
"[default=%default].")
parser.add_option("-b",
"--background-tsv-file",
dest="filename_background",
type="string",
help="filename with background genes to analyse "
"[default=%default].")
parser.add_option("-m",
"--min-counts",
dest="minimum_counts",
type="int",
help="minimum count - ignore all categories that have "
"fewer than # number of genes"
" [default=%default].")
parser.add_option("-o",
"--sort-order",
dest="sort_order",
type="choice",
choices=("fdr", "pvalue", "ratio"),
help="output sort order [default=%default].")
parser.add_option("--ontology",
dest="ontology",
type="string",
action="append",
help="go ontologies to analyze. Ontologies are tested "
"separately [default=%default].")
parser.add_option(
"-t",
"--threshold",
dest="threshold",
type="float",
help="significance threshold [>1.0 = all ]. If --fdr is set, this "
"refers to the fdr, otherwise it is a cutoff for p-values.")
parser.add_option("--filename-dump",
dest="filename_dump",
type="string",
help="dump GO category assignments into a flatfile "
"[default=%default].")
parser.add_option(
"--gene2name-map-tsv-file",
dest="filename_gene2name",
type="string",
help="optional filename mapping gene identifiers to gene names "
"[default=%default].")
parser.add_option(
"--filename-ontology",
dest="filename_ontology",
type="string",
help="filename with ontology in OBO format [default=%default].")
parser.add_option("--filename-input",
dest="filename_input",
type="string",
help="read GO category assignments from a flatfile "
"[default=%default].")
parser.add_option("--sample-size",
dest="sample",
type="int",
help="do sampling (with # samples) [default=%default].")
parser.add_option(
"--filename-output-pattern",
"--output-filename-pattern",
dest="output_filename_pattern",
type="string",
help="pattern with output filename pattern "
"(should contain: %(go)s and %(section)s ) [default=%default]")
parser.add_option("--fdr",
dest="fdr",
action="store_true",
help="calculate and filter by FDR default=%default].")
parser.add_option(
"--go2goslim",
dest="go2goslim",
action="store_true",
help="convert go assignments in STDIN to goslim assignments and "
"write to STDOUT [default=%default].")
parser.add_option("--gene-pattern",
dest="gene_pattern",
type="string",
help="pattern to transform identifiers to GO gene names "
"[default=%default].")
parser.add_option("--filename-map-slims",
dest="filename_map_slims",
type="string",
help="write mapping between GO categories and GOSlims "
"[default=%default].")
parser.add_option(
"--get-genes",
dest="get_genes",
type="string",
help="list all genes in the with a certain GOID [default=%default].")
parser.add_option(
"--strict",
dest="strict",
action="store_true",
help="require all genes in foreground to be part of background. "
"If not set, genes in foreground will be added to the background "
"[default=%default].")
parser.add_option(
"-q",
"--fdr-method",
dest="qvalue_method",
type="choice",
choices=("empirical", "storey", "BH"),
help="method to perform multiple testing correction by controlling "
"the fdr [default=%default].")
parser.add_option(
"--pairwise",
dest="compute_pairwise",
action="store_true",
help="compute pairwise enrichment for multiple gene lists. "
"[default=%default].")
# parser.add_option( "--fdr-lambda", dest="qvalue_lambda", type="float",
# help="fdr computation: lambda [default=%default]." )
# parser.add_option( "--qvalue-pi0-method", dest="qvalue_pi0_method", type="choice",
# choices = ("smoother", "bootstrap" ),
# help="fdr computation: method for estimating pi0 [default=%default]." )
parser.set_defaults(species=None,
filename_genes="-",
filename_background=None,
filename_slims=None,
minimum_counts=0,
ontology=[],
filename_dump=None,
sample=0,
fdr=False,
output_filename_pattern=None,
threshold=0.05,
filename_map_slims=None,
gene_pattern=None,
sort_order="ratio",
get_genes=None,
strict=False,
qvalue_method="empirical",
pairs_min_observed_counts=3,
compute_pairwise=False,
filename_gene2name=None)
(options, args) = E.Start(parser, add_database_options=True)
if options.go2goslim:
GO.convertGo2Goslim(options)
E.Stop()
sys.exit(0)
if options.fdr and options.sample == 0:
E.warn("fdr will be computed without sampling")
#############################################################
# dump GO
if options.filename_dump:
# set default orthologies to GO
if not options.ontology:
options.ontology = [
"biol_process", "mol_function", "cell_location"
]
E.info("dumping GO categories to %s" % (options.filename_dump))
dbhandle = connectToEnsembl(options)
outfile = IOTools.openFile(options.filename_dump, "w", create_dir=True)
GO.DumpGOFromDatabase(outfile, dbhandle, options)
outfile.close()
E.Stop()
sys.exit(0)
#############################################################
# read GO categories from file
if options.filename_input:
E.info("reading association of categories and genes from %s" %
(options.filename_input))
infile = IOTools.openFile(options.filename_input)
gene2gos, go2infos = GO.ReadGene2GOFromFile(infile)
infile.close()
if options.filename_gene2name:
E.info("reading gene identifier to gene name mapping from %s" %
options.filename_gene2name)
infile = IOTools.openFile(options.filename_gene2name)
gene2name = IOTools.readMap(infile, has_header=True)
infile.close()
E.info("read %i gene names for %i gene identifiers" %
(len(set(gene2name.values())), len(gene2name)))
else:
# use identity mapping
gene2name = dict([(x, x) for x in list(gene2gos.keys())])
#############################################################
# read GO ontology from file
if options.filename_ontology:
E.info("reading ontology from %s" % (options.filename_ontology))
infile = IOTools.openFile(options.filename_ontology)
ontology = GO.readOntology(infile)
infile.close()
def _g():
return collections.defaultdict(GO.GOInfo)
go2infos = collections.defaultdict(_g)
# substitute go2infos
for go in list(ontology.values()):
go2infos[go.mNameSpace][go.mId] = GO.GOInfo(go.mId,
go_type=go.mNameSpace,
description=go.mName)
#############################################################
# get foreground gene list
input_foreground, genelists = GO.ReadGeneLists(
options.filename_genes, gene_pattern=options.gene_pattern)
E.info("read %i genes for forground in %i gene lists" %
(len(input_foreground), len(genelists)))
#############################################################
# get background
if options.filename_background:
# nick - bug fix: background is the first tuple element from
# ReadGeneLists
input_background = GO.ReadGeneLists(
options.filename_background, gene_pattern=options.gene_pattern)[0]
E.info("read %i genes for background" % len(input_background))
else:
input_background = None
#############################################################
# sort out which ontologies to test
if not options.ontology:
if options.filename_input:
options.ontology = list(gene2gos.keys())
E.info("found %i ontologies: %s" %
(len(options.ontology), options.ontology))
summary = []
summary.append("\t".join(
("genelist", "ontology", "significant", "threshold", "ngenes",
"ncategories", "nmaps", "nforegound", "nforeground_mapped",
"nbackground", "nbackground_mapped", "nsample_counts",
"nbackground_counts", "psample_assignments",
"pbackground_assignments", "messages")) + "\n")
#############################################################
# get go categories for genes
for test_ontology in sorted(options.ontology):
# store results for aggregate output of multiple gene lists
all_results = []
all_significant_results = []
all_genelists_with_results = []
E.info("working on ontology %s" % test_ontology)
#############################################################
# get/read association of GO categories to genes
if options.filename_input:
gene2go, go2info = gene2gos[test_ontology], go2infos[test_ontology]
else:
E.info("reading data from database ...")
dbhandle.Connect(options)
gene2go, go2info = GO.ReadGene2GOFromDatabase(
dbhandle, test_ontology, options.database, options.species)
E.info("finished")
if len(go2info) == 0:
E.warn("could not find information for terms - "
"could be mismatch between ontologies")
ngenes, ncategories, nmaps, counts_per_category = GO.CountGO(gene2go)
E.info("assignments found: %i genes mapped to %i categories "
"(%i maps)" % (ngenes, ncategories, nmaps))
if options.minimum_counts > 0:
to_remove = set([
x for x, y in counts_per_category.items()
if y < options.minimum_counts
])
E.info("removing %i categories with less than %i genes" %
(len(to_remove), options.minimum_counts))
GO.removeCategories(gene2go, to_remove)
ngenes, ncategories, nmaps, counts_per_category = \
GO.CountGO(gene2go)
E.info("assignments after filtering: %i genes mapped "
"to %i categories (%i maps)" % (ngenes, ncategories, nmaps))
for genelist_name, foreground in sorted(genelists.items()):
msgs = []
E.info("processing %s with %i genes" %
(genelist_name, len(foreground)))
##################################################################
##################################################################
##################################################################
# build background - reconcile with foreground
##################################################################
if input_background is None:
background = list(gene2go.keys())
else:
background = list(input_background)
# nick - bug-fix backgorund included the foreground in a tuple.
# background is the first tuple element
missing = foreground.difference(set(background))
if options.strict:
assert len(missing) == 0, \
"%i genes in foreground but not in background: %s" % (
len(missing), str(missing))
else:
if len(missing) != 0:
E.warn("%i genes in foreground that are not in "
"background - added to background of %i" %
(len(missing), len(background)))
background.extend(missing)
E.info("(unfiltered) foreground=%i, background=%i" %
(len(foreground), len(background)))
# sort foreground and background, important for reproducibility
# under random seed
foreground = sorted(foreground)
background = sorted(background)
#############################################################
# sanity checks:
# are all of the foreground genes in the dataset
# missing = set(genes).difference( set(gene2go.keys()) )
# assert len(missing) == 0, "%i genes in foreground set without GO annotation: %s" % (len(missing), str(missing))
#############################################################
# read GO slims and map GO categories to GO slim categories
if options.filename_slims:
go_slims = GO.GetGOSlims(
IOTools.openFile(options.filename_slims, "r"))
if options.loglevel >= 1:
v = set()
for x in list(go_slims.values()):
for xx in x:
v.add(xx)
options.stdlog.write(
"# read go slims from %s: go=%i, slim=%i\n" %
(options.filename_slims, len(go_slims), len(v)))
if options.filename_map_slims:
if options.filename_map_slims == "-":
outfile = options.stdout
else:
outfile = IOTools.openFile(options.filename_map_slims,
"w")
outfile.write("GO\tGOSlim\n")
for go, go_slim in sorted(list(go_slims.items())):
outfile.write("%s\t%s\n" % (go, go_slim))
if outfile != options.stdout:
outfile.close()
gene2go = GO.MapGO2Slims(gene2go, go_slims, ontology=ontology)
if options.loglevel >= 1:
ngenes, ncategories, nmaps, counts_per_category = \
GO.CountGO(gene2go)
options.stdlog.write(
"# after go slim filtering: %i genes mapped to "
"%i categories (%i maps)\n" %
(ngenes, ncategories, nmaps))
#############################################################
# Just dump out the gene list
if options.get_genes:
fg, bg, ng = [], [], []
for gene, vv in list(gene2go.items()):
for v in vv:
if v.mGOId == options.get_genes:
if gene in genes:
fg.append(gene)
elif gene in background:
bg.append(gene)
else:
ng.append(gene)
# skip to next GO class
if not (bg or ng):
continue
options.stdout.write("# genes in GO category %s\n" %
options.get_genes)
options.stdout.write("gene\tset\n")
for x in sorted(fg):
options.stdout.write("%s\t%s\n" % ("fg", x))
for x in sorted(bg):
options.stdout.write("%s\t%s\n" % ("bg", x))
for x in sorted(ng):
options.stdout.write("%s\t%s\n" % ("ng", x))
E.info("nfg=%i, nbg=%i, nng=%i" % (len(fg), len(bg), len(ng)))
E.Stop()
sys.exit(0)
#############################################################
outfile = GO.getFileName(options,
go=test_ontology,
section='foreground',
set=genelist_name)
outfile.write("gene_id\n%s\n" % ("\n".join(sorted(foreground))))
if options.output_filename_pattern:
outfile.close()
outfile = GO.getFileName(options,
go=test_ontology,
section='background',
set=genelist_name)
# Jethro bug fix - see section 'build background' for assignment
outfile.write("gene_id\n%s\n" % ("\n".join(sorted(background))))
if options.output_filename_pattern:
outfile.close()
#############################################################
# do the analysis
go_results = GO.AnalyseGO(gene2go, foreground, background)
if len(go_results.mSampleGenes) == 0:
E.warn("%s: no genes with GO categories - analysis aborted" %
genelist_name)
continue
pairs = list(go_results.mResults.items())
#############################################################
# calculate fdr for each hypothesis
if options.fdr:
fdrs, samples, method = GO.computeFDRs(go_results, foreground,
background, options,
test_ontology, gene2go,
go2info)
for x, v in enumerate(pairs):
v[1].mQValue = fdrs[v[0]][0]
else:
fdrs, samples, method = {}, {}, None
msgs.append("fdr=%s" % method)
if options.sort_order == "fdr":
pairs.sort(key=lambda x: x[1].mQValue)
elif options.sort_order == "ratio":
pairs.sort(key=lambda x: x[1].mRatio)
elif options.sort_order == "pvalue":
pairs.sort(key=lambda x: x[1].mPValue)
#############################################################
#############################################################
#############################################################
# output the full result
outfile = GO.getFileName(options,
go=test_ontology,
section='overall',
set=genelist_name)
GO.outputResults(outfile,
pairs,
go2info,
options,
fdrs=fdrs,
samples=samples)
if options.output_filename_pattern:
outfile.close()
#############################################################
#############################################################
#############################################################
# filter significant results and output
filtered_pairs = GO.selectSignificantResults(pairs, fdrs, options)
nselected = len(filtered_pairs)
nselected_up = len([x for x in filtered_pairs if x[1].mRatio > 1])
nselected_down = len(
[x for x in filtered_pairs if x[1].mRatio < 1])
assert nselected_up + nselected_down == nselected
outfile = GO.getFileName(options,
go=test_ontology,
section='results',
set=genelist_name)
GO.outputResults(outfile,
filtered_pairs,
go2info,
options,
fdrs=fdrs,
samples=samples)
if options.output_filename_pattern:
outfile.close()
#############################################################
#############################################################
#############################################################
# save results for multi-gene-list analysis
all_results.append(pairs)
all_significant_results.append(filtered_pairs)
all_genelists_with_results.append(genelist_name)
#############################################################
#############################################################
#############################################################
# output parameters
ngenes, ncategories, nmaps, counts_per_category = \
GO.CountGO(gene2go)
outfile = GO.getFileName(options,
go=test_ontology,
section='parameters',
set=genelist_name)
nbackground = len(background)
if nbackground == 0:
nbackground = len(go_results.mBackgroundGenes)
outfile.write(
"# input go mappings for gene list '%s' and category '%s'\n" %
(genelist_name, test_ontology))
outfile.write("parameter\tvalue\tdescription\n")
outfile.write("mapped_genes\t%i\tmapped genes\n" % ngenes)
outfile.write("mapped_categories\t%i\tmapped categories\n" %
ncategories)
outfile.write("mappings\t%i\tmappings\n" % nmaps)
outfile.write("genes_in_fg\t%i\tgenes in foreground\n" %
len(foreground))
outfile.write(
"genes_in_fg_with_assignment\t%i\tgenes in foreground with GO assignments\n"
% (len(go_results.mSampleGenes)))
outfile.write("genes_in_bg\t%i\tinput background\n" % nbackground)
outfile.write(
"genes_in_bg_with_assignment\t%i\tgenes in background with GO assignments\n"
% (len(go_results.mBackgroundGenes)))
outfile.write("associations_in_fg\t%i\tassociations in sample\n" %
go_results.mSampleCountsTotal)
outfile.write(
"associations_in_bg\t%i\tassociations in background\n" %
go_results.mBackgroundCountsTotal)
outfile.write(
"percent_genes_in_fg_with_association\t%s\tpercent genes in sample with GO assignments\n"
% (IOTools.prettyPercent(len(go_results.mSampleGenes),
len(foreground), "%5.2f")))
outfile.write(
"percent_genes_in_bg_with_associations\t%s\tpercent genes background with GO assignments\n"
% (IOTools.prettyPercent(len(go_results.mBackgroundGenes),
nbackground, "%5.2f")))
outfile.write("significant\t%i\tsignificant results reported\n" %
nselected)
outfile.write(
"significant_up\t%i\tsignificant up-regulated results reported\n"
% nselected_up)
outfile.write(
"significant_down\t%i\tsignificant up-regulated results reported\n"
% nselected_down)
outfile.write("threshold\t%6.4f\tsignificance threshold\n" %
options.threshold)
if options.output_filename_pattern:
outfile.close()
summary.append("\t".join(
map(str, (genelist_name, test_ontology, nselected,
options.threshold, ngenes, ncategories, nmaps,
len(foreground), len(go_results.mSampleGenes),
nbackground, len(go_results.mBackgroundGenes),
go_results.mSampleCountsTotal,
go_results.mBackgroundCountsTotal,
IOTools.prettyPercent(len(go_results.mSampleGenes),
len(foreground), "%5.2f"),
IOTools.prettyPercent(
len(go_results.mBackgroundGenes), nbackground,
"%5.2f"), ",".join(msgs)))) + "\n")
#############################################################
#############################################################
#############################################################
# output the fg patterns
outfile = GO.getFileName(options,
go=test_ontology,
section='withgenes',
set=genelist_name)
GO.outputResults(outfile,
pairs,
go2info,
options,
fdrs=fdrs,
samples=samples,
gene2go=gene2go,
foreground=foreground,
gene2name=gene2name)
if options.output_filename_pattern:
outfile.close()
if len(genelists) > 1:
###################################################################
# output various summary files
# significant results
GO.outputMultipleGeneListResults(all_significant_results,
all_genelists_with_results,
test_ontology,
go2info,
options,
section='significant')
# all results
GO.outputMultipleGeneListResults(all_results,
all_genelists_with_results,
test_ontology,
go2info,
options,
section='all')
if options.compute_pairwise:
GO.pairwiseGOEnrichment(all_results,
all_genelists_with_results,
test_ontology, go2info, options)
outfile_summary = options.stdout
outfile_summary.write("".join(summary))
E.Stop()