def main(mcoolfile,outdir,filename,window,cutoff,binsize):
f = mcoolfile
#Get the list of resolutions in the mcool file
cooler_list = cooler.io.ls(f)
old_version= False
if not any([res for res in cooler_list if '/resolutions/' in res]): #gets the resolutions from a file in a older version of cooler
old_version = True
binsize_list = []
for res in cooler_list:
cooler_path = str(f)+'::'+ res
c = cooler.Cooler(cooler_path)
binsize_list.append(int(c.binsize))
else:
binsize_list = []
for res in cooler_list:
binsize_list.append(int(res.split('/')[-1]))
# Check the input parameters
if binsize == -1:
binsize = min(binsize_list)
else:
if binsize in binsize_list:
if window % binsize != 0:
print("Error: Window size must be multiple of binsize")
sys.exit()
else:
print("Error: This binsize is not available in this mcool file. This is the list of binsizes availables:")
print(binsize_list)
sys.exit()
# Creates a cooler object
if old_version:
res_list = []
for res in cooler_list:
res_list.append(int(res.split('/')[-1]))
res_index = max(res_list)
cooler_path = str(f) + '::' + str(res_index)
else:
cooler_path = str(f) + '::' + cooler_list[binsize_list.index(binsize)]
c = cooler.Cooler(cooler_path)
print(c)
# Gets the chromsizes
chromsizes=pd.Series(c.chroms()[:]['length'].values, index=c.chroms()[:]['name'].values)
#Getting insulating boundaries
insul = find_insulating_boundaries(c,balance='weight',window_bp=window,min_dist_bad_bin=2)
#Convert to BigWig
bioframe.to_bigwig(insul, chromsizes,
f'/{outdir}/{filename}.bw',
f'log2_insulation_score_{window}')
def eigs_cis(
cool_path,
phasing_track,
view,
n_eigs,
clr_weight_name,
ignore_diags,
verbose,
out_prefix,
bigwig,
):
"""
Perform eigen value decomposition on a cooler matrix to calculate
compartment signal by finding the eigenvector that correlates best with the
phasing track.
COOL_PATH : the paths to a .cool file with a balanced Hi-C map. Use the
'::' syntax to specify a group path in a multicooler file.
TRACK_PATH : the path to a BedGraph-like file that stores phasing track as
track-name named column.
BedGraph-like format assumes tab-separated columns chrom, start, stop and
track-name.
"""
clr = cooler.Cooler(cool_path)
if phasing_track is not None:
# TODO: This all needs to be refactored into a more generic tabular file parser
# Needs to handle stdin case too.
track_path, col = phasing_track
buf, names = sniff_for_header(track_path)
if names is None:
if not isinstance(col, int):
raise click.BadParameter(
"No header found. "
'Cannot find "{}" column without a header.'.format(col))
track_name = "ref"
kwargs = dict(
header=None,
usecols=[0, 1, 2, col],
names=["chrom", "start", "end", track_name],
)
else:
if isinstance(col, int):
try:
col = names[col]
except IndexError:
raise click.BadParameter(
'Column #{} not compatible with header "{}".'.format(
col, ",".join(names)))
else:
if col not in names:
raise click.BadParameter(
'Column "{}" not found in header "{}"'.format(
col, ",".join(names)))
track_name = col
kwargs = dict(header="infer",
usecols=["chrom", "start", "end", track_name])
track_df = pd.read_table(buf,
dtype={
"chrom": str,
"start": np.int64,
"end": np.int64,
track_name: np.float64,
},
comment="#",
verbose=verbose,
**kwargs)
phasing_track = track_df
# define view for cis compartment-calling
# use input "view" BED file or all chromosomes mentioned in "track":
if view is None:
cooler_view_df = make_cooler_view(clr)
view_df = cooler_view_df
else:
view_df = read_viewframe_from_file(view, clr, check_sorting=True)
# TODO: Add check that view_df has the same bins as track
eigvals, eigvec_table = eigdecomp.eigs_cis(
clr=clr,
phasing_track=phasing_track,
view_df=view_df,
n_eigs=n_eigs,
clr_weight_name=clr_weight_name,
ignore_diags=ignore_diags,
clip_percentile=99.9,
sort_metric=None,
)
# Output
eigvals.to_csv(out_prefix + ".cis" + ".lam.txt", sep="\t", index=False)
eigvec_table.to_csv(out_prefix + ".cis" + ".vecs.tsv",
sep="\t",
index=False)
if bigwig:
bioframe.to_bigwig(
eigvec_table,
clr.chromsizes,
out_prefix + ".cis" + ".bw",
value_field="E1",
)
def insulation(
in_path,
window,
output,
view,
ignore_diags,
clr_weight_name,
min_frac_valid_pixels,
min_dist_bad_bin,
threshold,
window_pixels,
append_raw_scores,
chunksize,
verbose,
bigwig,
):
"""
Calculate the diamond insulation scores and call insulating boundaries.
IN_PATH : The paths to a .cool file with a balanced Hi-C map.
WINDOW : The window size for the insulation score calculations.
Multiple space-separated values can be provided.
By default, the window size must be provided in units of bp.
When the flag --window-pixels is set, the window sizes must
be provided in units of pixels instead.
"""
clr = cooler.Cooler(in_path)
# Create view:
cooler_view_df = make_cooler_view(clr)
if view is None:
# full chromosomes:
view_df = cooler_view_df
else:
# read view_df dataframe, and verify against cooler
view_df = read_viewframe_from_file(view, clr, check_sorting=True)
# Read list with windows:
if window_pixels:
window = [win * clr.info["bin-size"] for win in window]
ins_table = api.insulation.insulation(
clr,
view_df=view_df,
window_bp=window,
ignore_diags=ignore_diags,
clr_weight_name=clr_weight_name if clr_weight_name else None,
min_frac_valid_pixels=min_frac_valid_pixels,
min_dist_bad_bin=min_dist_bad_bin,
threshold=threshold,
append_raw_scores=append_raw_scores,
chunksize=chunksize,
verbose=verbose,
)
# output to file if specified:
if output:
ins_table.to_csv(output, sep="\t", index=False, na_rep="nan")
# or print into stdout otherwise:
else:
print(ins_table.to_csv(sep="\t", index=False, na_rep="nan"))
# Write the insulation track as a bigwig:
if bigwig:
for w in window:
bioframe.to_bigwig(
ins_table,
clr.chromsizes,
output + "." + str(w) + ".bw",
value_field=f"log2_insulation_score_{w}",
)
def call_compartments(cool_path, reference_track, contact_type, n_eigs,
verbose, out_prefix, bigwig):
"""
Perform eigen value decomposition on a cooler matrix to calculate
compartment signal by finding the eigenvector that correlates best with the
phasing track.
COOL_PATH : the paths to a .cool file with a balanced Hi-C map.
TRACK_PATH : the path to a BedGraph-like file that stores phasing track as
track-name named column.
BedGraph-like format assumes tab-separated columns chrom, start, stop and
track-name.
"""
clr = cooler.Cooler(cool_path)
if reference_track is not None:
# TODO: This all needs to be refactored into a more generic tabular file parser
# Needs to handle stdin case too.
track_path, col = reference_track
buf, names = sniff_for_header(track_path)
if names is None:
if not isinstance(col, int):
raise click.BadParameter(
"No header found. "
'Cannot find "{}" column without a header.'.format(col))
track_name = "ref"
kwargs = dict(
header=None,
usecols=[0, 1, 2, col],
names=["chrom", "start", "end", track_name],
)
else:
if isinstance(col, int):
try:
col = names[col]
except IndexError:
raise click.BadParameter(
'Column #{} not compatible with header "{}".'.format(
col, ",".join(names)))
else:
if col not in names:
raise click.BadParameter(
'Column "{}" not found in header "{}"'.format(
col, ",".join(names)))
track_name = col
kwargs = dict(header="infer",
usecols=["chrom", "start", "end", track_name])
track_df = pd.read_table(buf,
dtype={
"chrom": str,
"start": np.int64,
"end": np.int64,
track_name: np.float64,
},
comment="#",
verbose=verbose,
**kwargs)
# we need to merge phasing track DataFrame with the cooler bins to get
# a DataFrame with phasing info aligned and validated against bins inside of
# the cooler file.
track = pd.merge(left=clr.bins()[:],
right=track_df,
how="left",
on=["chrom", "start", "end"])
# sanity check would be to check if len(bins) becomes > than nbins ...
# that would imply there was something in the track_df that didn't match
# ["chrom", "start", "end"] - keys from the c.bins()[:] .
if len(track) > len(clr.bins()):
ValueError(
"There is something in the {} that ".format(track_path) +
"couldn't be merged with cooler-bins {}".format(cool_path))
else:
track = clr.bins()[["chrom", "start", "end"]][:]
track_name = None
# it's contact_type dependent:
if contact_type == "cis":
eigvals, eigvec_table = eigdecomp.cooler_cis_eig(
clr=clr,
bins=track,
regions=None,
n_eigs=n_eigs,
phasing_track_col=track_name,
clip_percentile=99.9,
sort_metric=None,
)
elif contact_type == "trans":
eigvals, eigvec_table = eigdecomp.cooler_trans_eig(
clr=clr,
bins=track,
n_eigs=n_eigs,
partition=None,
phasing_track_col=track_name,
sort_metric=None,
)
# Output
eigvals.to_csv(out_prefix + "." + contact_type + ".lam.txt",
sep="\t",
index=False)
eigvec_table.to_csv(out_prefix + "." + contact_type + ".vecs.tsv",
sep="\t",
index=False)
if bigwig:
bioframe.to_bigwig(
eigvec_table,
clr.chromsizes,
out_prefix + "." + contact_type + ".bw",
value_field="E1",
)
def save_bigwig(vectors, savepath, genome, columns=['E1', 'E2', 'E3']):
chroms = fetch_chromsizes(genome)
for item in columns:
save = savepath+'.{}.bw'.format(item)
create_dir(save)
to_bigwig(vectors, chroms, save, value_field=item)
def main(mcoolfile, outdir, filename, binsize, window, bweak, bstrong, cutoff,
pixels_frac):
f = mcoolfile
# Get the list of resolutions in the mcool file
cooler_list = cooler.fileops.list_coolers(f)
old_version = False
# gets the resolutions from a file in a older version of cooler
if not any([res for res in cooler_list if '/resolutions/' in res]):
old_version = True
binsize_list = []
for res in cooler_list:
cooler_path = str(f) + '::' + res
c = cooler.Cooler(cooler_path)
binsize_list.append(int(c.binsize))
else:
binsize_list = []
for res in cooler_list:
binsize_list.append(int(res.split('/')[-1]))
# Check the input parameters
if binsize == -1:
binsize = min(binsize_list)
else:
if binsize in binsize_list:
if window % binsize != 0:
print("Error: Window size must be multiple of binsize")
sys.exit()
else:
print(
"Error: This binsize is not available in this mcool file. This is the list of binsizes availables:"
)
print(binsize_list)
sys.exit()
# Creates a cooler object
if old_version:
res_list = []
for res in cooler_list:
res_list.append(int(res.split('/')[-1]))
res_index = max(res_list)
cooler_path = str(f) + '::' + str(res_index)
else:
cooler_path = str(f) + '::' + cooler_list[binsize_list.index(binsize)]
c = cooler.Cooler(cooler_path)
print(c)
# Gets the chromsizes
chromsizes = pd.Series(c.chroms()[:]['length'].values,
index=c.chroms()[:]['name'].values)
# calculate insulation score
ins_table = calculate_insulation_score(c, window)
# Find boundaries
ins_table = find_boundaries(ins_table, pixels_frac, cutoff)
# Filter out insulation score in which pixels are less than 66% of the max number of pixels
icutoff = pow((window / binsize), 2) * pixels_frac
ins_table_filtered = ins_table[
ins_table[f'n_valid_pixels_{window}'] >= icutoff]
# Convert to BigWig
bioframe.to_bigwig(ins_table_filtered, chromsizes,
f'{outdir}/{filename}.bw',
f'log2_insulation_score_{window}')
# Classify the boundaries as strong and week
# strong boundaries >= 0.5, weak 0.2>= and < 0.5
boun_table = ins_table_filtered[
ins_table_filtered[f'boundary_strength_{window}'] >= bweak].copy()
boun_table['boundary_classification'] = np.where(
boun_table[f'boundary_strength_{window}'] >= bstrong, 'Strong', 'Weak')
# Filter boundaries abover thresholds
columns = [
'chrom', 'start', 'end', 'boundary_classification',
f'boundary_strength_{window}'
]
boun_table.to_csv(f'{outdir}/{filename}_boundaries.bed',
sep='\t',
header=None,
index=False,
columns=columns)
for chrom in cool.chromnames[0:22]]}
#supports
# Computing eigenvealues and eigenvectors
lams, vectors = eigdecomp.cooler_cis_eig(cool, genes, regions=supports['cis'],
phasing_track_col='gene_count',
sort_metric='spearmanr')
# cooler_cis_eig sorts eigenvectors by decreasing Spearman correlation with the phasing track
# (gene count in this case). In the past, people have used the eigenvector associated with the max
# eigenvalue. It is worth considering situations where these two sorting process differ.
# Output
contact_type='cis'
lams.to_csv(out_prefix + '.' + contact_type +'.'+ res+ '.lam.txt', sep='\t', index=False)
vectors.to_csv(out_prefix + '.' + contact_type +'.'+ res+'.vecs.tsv', sep='\t', index=False)
bioframe.to_bigwig(vectors,cool.chromsizes,out_prefix +'.' + contact_type + '.'+ res+ '.bw',value_field='E1')
exp = {}
#Cis Expected
with mp.Pool(10) as p:
result = expected.diagsum(cool, supports['cis'],
transforms={'balanced': lambda p: p['count']*p['weight1']*p['weight2']},
ignore_diags=2, map=p.map)
result = pd.concat([result[arm] for arm in supports['cis']],
keys=[(arm[0], f'{arm[0]}:{arm[1]}-{arm[2]}') for arm in supports['cis']],
names=['chrom', 'region'])
result = result.groupby(['chrom','diag']).sum()
result['balanced.avg'] = result['balanced.sum'] / result['n_valid']
result = result.reset_index()
exp['cis'] = result
def call_compartments(
cool_path,
reference_track,
regions,
contact_type,
n_eigs,
verbose,
out_prefix,
bigwig,
):
"""
Perform eigen value decomposition on a cooler matrix to calculate
compartment signal by finding the eigenvector that correlates best with the
phasing track.
COOL_PATH : the paths to a .cool file with a balanced Hi-C map. Use the
'::' syntax to specify a group path in a multicooler file.
TRACK_PATH : the path to a BedGraph-like file that stores phasing track as
track-name named column.
BedGraph-like format assumes tab-separated columns chrom, start, stop and
track-name.
"""
clr = cooler.Cooler(cool_path)
if reference_track is not None:
# TODO: This all needs to be refactored into a more generic tabular file parser
# Needs to handle stdin case too.
track_path, col = reference_track
buf, names = sniff_for_header(track_path)
if names is None:
if not isinstance(col, int):
raise click.BadParameter(
"No header found. "
'Cannot find "{}" column without a header.'.format(col))
track_name = "ref"
kwargs = dict(
header=None,
usecols=[0, 1, 2, col],
names=["chrom", "start", "end", track_name],
)
else:
if isinstance(col, int):
try:
col = names[col]
except IndexError:
raise click.BadParameter(
'Column #{} not compatible with header "{}".'.format(
col, ",".join(names)))
else:
if col not in names:
raise click.BadParameter(
'Column "{}" not found in header "{}"'.format(
col, ",".join(names)))
track_name = col
kwargs = dict(header="infer",
usecols=["chrom", "start", "end", track_name])
track_df = pd.read_table(buf,
dtype={
"chrom": str,
"start": np.int64,
"end": np.int64,
track_name: np.float64,
},
comment="#",
verbose=verbose,
**kwargs)
# we need to merge phasing track DataFrame with the cooler bins to get
# a DataFrame with phasing info aligned and validated against bins inside of
# the cooler file.
track = pd.merge(left=clr.bins()[:],
right=track_df,
how="left",
on=["chrom", "start", "end"])
# sanity check would be to check if len(bins) becomes > than nbins ...
# that would imply there was something in the track_df that didn't match
# ["chrom", "start", "end"] - keys from the c.bins()[:] .
if len(track) > len(clr.bins()):
ValueError(
"There is something in the {} that ".format(track_path) +
"couldn't be merged with cooler-bins {}".format(cool_path))
else:
# use entire bin-table from cooler, when reference-track is not provided:
track = clr.bins()[["chrom", "start", "end"]][:]
track_name = None
# define regions for cis compartment-calling
# use input "regions" BED file or all chromosomes mentioned in "track":
if regions is None:
# use full chromosomes referred to in the track :
track_chroms = track["chrom"].unique()
cis_regions_table = bioframe.parse_regions(track_chroms,
clr.chromsizes)
cis_regions_table["name"] = cis_regions_table["chrom"]
else:
if contact_type == "trans":
raise NotImplementedError(
"Regions not yet supported with trans contact type")
# Flexible reading of the regions table:
regions_buf, names = sniff_for_header(regions)
cis_regions_table = pd.read_csv(regions_buf, sep="\t", header=None)
if cis_regions_table.shape[1] not in (3, 4):
raise ValueError(
"The region file does not have three or four tab-delimited columns."
"We expect a bed file with columns chrom, start, end, and optional name"
)
if cis_regions_table.shape[1] == 4:
cis_regions_table = cis_regions_table.rename(columns={
0: "chrom",
1: "start",
2: "end",
3: "name"
})
cis_regions_table = bioframe.parse_regions(cis_regions_table)
else:
cis_regions_table = cis_regions_table.rename(columns={
0: "chrom",
1: "start",
2: "end"
})
cis_regions_table = bioframe.parse_regions(cis_regions_table)
# make sure custom regions are compatible with the track:
track_chroms = track["chrom"].unique()
cis_regions_table = cis_regions_table[cis_regions_table["chrom"].isin(
track_chroms)].reset_index(drop=True)
# it's contact_type dependent:
if contact_type == "cis":
eigvals, eigvec_table = eigdecomp.cooler_cis_eig(
clr=clr,
bins=track,
regions=cis_regions_table,
n_eigs=n_eigs,
phasing_track_col=track_name,
clip_percentile=99.9,
sort_metric=None,
)
elif contact_type == "trans":
eigvals, eigvec_table = eigdecomp.cooler_trans_eig(
clr=clr,
bins=track,
n_eigs=n_eigs,
partition=None,
phasing_track_col=track_name,
sort_metric=None,
)
# Output
eigvals.to_csv(out_prefix + "." + contact_type + ".lam.txt",
sep="\t",
index=False)
eigvec_table.to_csv(out_prefix + "." + contact_type + ".vecs.tsv",
sep="\t",
index=False)
if bigwig:
bioframe.to_bigwig(
eigvec_table,
clr.chromsizes,
out_prefix + "." + contact_type + ".bw",
value_field="E1",
)
