def make_motif_logo_from_index(tcrsampler,
ind,
clone_df,
centroid,
cdr3_name='cdr3_b_aa',
v_name='v_b_gene',
gene_names=['v_b_gene', 'j_b_gene']):
"""
make motif logo from a specific index
"""
dfnode = clone_df.iloc[ind, :].copy()
dfnode[gene_names[0]] = dfnode[gene_names[0]].apply(lambda x: allele_01(x))
dfnode[gene_names[1]] = dfnode[gene_names[1]].apply(lambda x: allele_01(x))
gene_usage = dfnode.groupby(gene_names).size()
sampled_rep = tcrsampler.sample(
gene_usage.reset_index().to_dict('split')['data'],
flatten=True,
depth=100)
sampled_rep = [x for x in sampled_rep if x is not None]
motif, stat = compute_pal_motif(seqs=dfnode[cdr3_name],
refs=sampled_rep,
centroid=centroid)
svg = svg_logo(motif, return_str=True)
motif_raw, _ = compute_pal_motif(seqs=dfnode[cdr3_name], centroid=centroid)
svg_raw = svg_logo(motif_raw, return_str=True)
return svg, svg_raw
def motif_creation_human_betas():
import re
import pandas as pd
from tcrdist.repertoire import TCRrep
import palmotif
from tcrdist.pgen import OlgaModel
oma = OlgaModel(recomb_type="VJ", chain_folder="human_T_alpha")
from tcrdist.pgen import OlgaModel
omb = OlgaModel(recomb_type="VDJ", chain_folder="human_T_beta")
df = pd.read_csv("dash_human.csv")
tr = TCRrep(cell_df=df,
organism='human',
chains=['alpha', 'beta'],
db_file='alphabeta_gammadelta_db.tsv')
from tcrdist.adpt_funcs import get_basic_centroids
get_basic_centroids(tr, max_dist=75)
with open("test_3.svg", 'w') as oh:
oh.write('<body>')
for i, r in tr.centroids_df.iterrows():
if len(r['neighbors']) < 5:
break
seqs = tr.clone_df.iloc[r['neighbors'], ]['cdr3_b_aa'].to_list()
gene_usages = tr.clone_df.iloc[r['neighbors'], ][[
'v_b_gene', 'j_b_gene'
]].value_counts().reset_index().to_dict('split')['data']
depth = 3
refs = flatten([
omb.gen_cdr3s(allele_01(v), allele_01(j), i * depth)
for v, j, i in combos_alpha
])
refs = [x for x in refs if x is not None]
matrix, stats = palmotif.compute_pal_motif(seqs=seqs,
refs=refs,
centroid=r['cdr3_b_aa'])
matrix_raw, _ = palmotif.compute_pal_motif(seqs=seqs,
centroid=r['cdr3_b_aa'])
refs.append(r['cdr3_b_aa'])
matrix_bkgd, _ = palmotif.compute_pal_motif(
seqs=refs, centroid=r['cdr3_b_aa'])
svgs = [
palmotif.svg_logo(matrix, 'test.svg', return_str=True),
palmotif.svg_logo(matrix_raw, 'test.svg', return_str=True),
palmotif.svg_logo(matrix_bkgd, 'test.svg', return_str=True)
]
[oh.write(f"{s}<div></div>\n") for s in svgs]
oh.write('<div></div>')
oh.write(str(r))
oh.write('<div></div>')
oh.write('</body>')
def _discover_motif_in_cluster(self, tcr_rep, index, row, negative_examples=None) -> Tuple[List[ReportOutput], List[ReportOutput]]:
from tcrdist.adpt_funcs import get_centroid_seq
from tcrdist.summarize import _select
from palmotif import compute_pal_motif
from palmotif import svg_logo
dfnode = tcr_rep.clone_df.iloc[row['neighbors_i'],]
figure_outputs, table_outputs = [], []
logging.info(f"{TCRdistMotifDiscovery.__name__}: in cluster {index+1}, there are {dfnode.shape[0]} neighbors.")
for chain in ['a', 'b']:
if dfnode.shape[0] > 2:
centroid, *_ = get_centroid_seq(df=dfnode)
else:
centroid = dfnode[f'cdr3_{chain}_aa'].to_list()[0]
motif, stat = compute_pal_motif(seqs=_select(df=tcr_rep.clone_df, iloc_rows=row['neighbors_i'], col=f'cdr3_{chain}_aa'),
centroid=centroid, refs=negative_examples[chain] if self.use_reference_sequences else None)
figure_path = self.result_path / f"motif_{chain}_{index + 1}.svg"
svg_logo(motif, filename=figure_path)
motif_data_path = self.result_path / f"motif_{chain}_{index + 1}.csv"
motif.to_csv(motif_data_path)
figure_outputs.append(ReportOutput(figure_path, f'Motif {index + 1} ({Chain.get_chain(chain.upper()).name.lower()} chain)'))
table_outputs.append(ReportOutput(motif_data_path, f'motif {index + 1} ({Chain.get_chain(chain.upper()).name.lower()} chain) csv data'))
return figure_outputs, table_outputs
def _index_to_matrix(ind,
clone_df,
pwmat=None,
col='cdr3_b_aa',
centroid=None):
"""
Example
-------
"""
dfnode = clone_df.iloc[ind, ].copy()
seqs = dfnode[col].to_list()
if centroid is None:
pwnode = pwmat[ind, :][:, ind].copy()
iloc_idx = pwnode.sum(axis=0).argmin()
centroid = dfnode[col].to_list()[iloc_idx]
matrix, stats = compute_pal_motif(seqs=seqs, centroid=centroid)
return matrix
def test_gallery_hdiff():
"""
All imports are provided here, and are repeated
step-wise below, for clarity, and for
module cut-and-paste. This example
performs paired alpha-beta analysis,
but code blocks can be used for single
chain analysis as well.
"""
import pandas as pd
from tcrdist.repertoire import TCRrep
from tcrdist.rep_diff import hcluster_diff, member_summ
from tcrsampler.sampler import TCRsampler
from tcrdist.adpt_funcs import get_centroid_seq
from tcrdist.summarize import _select
from palmotif import compute_pal_motif, svg_logo
from hierdiff import plot_hclust_props
"""
Load a subset of data that contains paired alpha-beta
chain mouse TCR receptors that recognized
the PA or PB1 epitopes (present in mouse influenza).
"""
import pandas as pd
df = pd.read_csv("dash.csv")
conditional = df['epitope'].apply( lambda x: x in ['PA','PB1'])
"""
For illustrative/testing purposes, randomly subset the data to include
only 100 clones. Increase for more informative plot.
"""
df = df[conditional].\
reset_index(drop = True).\
sample(100, random_state = 3).\
reset_index(drop = True).\
copy()
"""
Load DataFrame into TCRrep instance,
which automatically computes attributes:
1. .clone_df DataFrame
2. .pw_beta nd.array
3. .pw_alpha nd.array
"""
from tcrdist.repertoire import TCRrep
tr = TCRrep(cell_df = df,
organism = 'mouse',
chains = ['beta','alpha'],
db_file = 'alphabeta_gammadelta_db.tsv')
"""
Apply hcluster_diff, which hierarchically clusters.
Note
----
pwmat could easily be tr.pw_beta or tr.pw_alpha if
clustering should be done on a single chain.
"""
from tcrdist.rep_diff import hcluster_diff
tr.hcluster_df, tr.Z =\
hcluster_diff(clone_df = tr.clone_df,
pwmat = tr.pw_beta + tr.pw_alpha,
x_cols = ['epitope'],
count_col = 'count')
"""
Load a custom background, mouse appropriate dataset to sample CDR3s
according to the V and J gene usage frequencies observed in each node.
See the tcrsampler package for more details
(https://github.com/kmayerb/tcrsampler/blob/master/docs/getting_default_backgrounds.md)
"""
from tcrsampler.sampler import TCRsampler
t = TCRsampler()
t.download_background_file("ruggiero_mouse_sampler.zip")
tcrsampler_beta = TCRsampler(default_background = 'ruggiero_mouse_beta_t.tsv.sampler.tsv')
tcrsampler_alpha = TCRsampler(default_background = 'ruggiero_mouse_alpha_t.tsv.sampler.tsv')
"""
Add an SVG graphic to every node of the tree
aligned to the cluster centroid.
"""
from tcrdist.adpt_funcs import get_centroid_seq
from tcrdist.summarize import _select
from palmotif import compute_pal_motif, svg_logo
"""Beta Chain"""
svgs_beta = list()
for i,r in tr.hcluster_df.iterrows():
dfnode = tr.clone_df.iloc[r['neighbors_i'],]
if dfnode.shape[0] > 2:
centroid, *_ = get_centroid_seq(df = dfnode)
else:
centroid = dfnode['cdr3_b_aa'].to_list()[0]
print(f"BETA-CHAIN: {centroid}")
gene_usage_beta = dfnode.groupby(['v_b_gene','j_b_gene']).size()
sampled_rep = tcrsampler_beta.sample( gene_usage_beta.reset_index().to_dict('split')['data'],
flatten = True, depth = 10)
sampled_rep = [x for x in sampled_rep if x is not None]
motif, stat = compute_pal_motif(
seqs = _select(df = tr.clone_df,
iloc_rows = r['neighbors_i'],
col = 'cdr3_b_aa'),
refs = sampled_rep,
centroid = centroid)
svgs_beta.append(svg_logo(motif, return_str= True))
"""Add Beta SVG graphics to hcluster_df"""
tr.hcluster_df['svg_beta'] = svgs_beta
"""Alpha Chain"""
svgs_alpha = list()
for i,r in tr.hcluster_df.iterrows():
dfnode = tr.clone_df.iloc[r['neighbors_i'],]
if dfnode.shape[0] > 2:
centroid, *_ = get_centroid_seq(df = dfnode)
else:
centroid = dfnode['cdr3_a_aa'].to_list()[0]
print(f"ALPHA-CHAIN: {centroid}")
gene_usage_alpha = dfnode.groupby(['v_a_gene','j_a_gene']).size()
sampled_rep = tcrsampler_alpha.sample( gene_usage_alpha.reset_index().to_dict('split')['data'],
flatten = True, depth = 10)
sampled_rep = [x for x in sampled_rep if x is not None]
motif, stat = compute_pal_motif(
seqs = _select(df = tr.clone_df,
iloc_rows = r['neighbors_i'],
col = 'cdr3_a_aa'),
refs = sampled_rep,
centroid = centroid)
svgs_alpha.append(svg_logo(motif, return_str= True))
"""Add Alpha SVG graphics to hcluster_df"""
tr.hcluster_df['svg_alpha'] = svgs_alpha
"""
Produce summary information for tooltips.
For instance, describe percentage of TCRs with
a given epitope at a given node.
"""
res_summary = member_summ( res_df = tr.hcluster_df,
clone_df = tr.clone_df,
addl_cols=['epitope'])
tr.hcluster_df_detailed = \
pd.concat([tr.hcluster_df, res_summary], axis = 1)
"""
Write D3 html for interactive denogram graphic.
Specify desired tooltips.
"""
from hierdiff import plot_hclust_props
html = plot_hclust_props(tr.Z,
title='PA Epitope Example',
res=tr.hcluster_df_detailed,
tooltip_cols=['cdr3_b_aa','v_b_gene', 'j_b_gene','svg_alpha','svg_beta'],
alpha=0.00001, colors = ['blue','gray'],
alpha_col='pvalue')
with open('hierdiff_example_PA_v_PB1.html', 'w') as fh:
fh.write(html)
def test_quick_pipeline_with_fragmented_compute():
"""
How can I used tcrdist3 to test for TCRs that may HLA restricted.
"""
import os
import pandas as pd
import numpy as np
from scipy import sparse
from tcrdist.repertoire import TCRrep
from tcrdist.rep_funcs import compute_pw_sparse_out_of_memory
f = 'mira_epitope_67_382_APHGVVFL_APHGVVFLHV_GVVFLHVTY_VVFLHVTYV.tcrdist3.csv'
f = os.path.join('tcrdist','data','covid19',f)
assert os.path.isfile(f)
df = pd.read_csv(f)
df = df[['subject', 'cell_type', 'v_b_gene', 'j_b_gene', 'cdr3_b_aa', 'cdr3_b_nucseq', 'cohort', 'hla-a', 'hla-a_1','hla-b', 'hla-b_1']]
tr = TCRrep(cell_df = df,
organism = 'human',
chains = ['beta'],
db_file = 'alphabeta_gammadelta_db.tsv',
compute_distances = False,
store_all_cdr = False)
from tcrdist.rep_funcs import compute_pw_sparse_out_of_memory
S, fragments = compute_pw_sparse_out_of_memory( tr = tr,
row_size = 100,
pm_processes = 2,
pm_pbar = True,
max_distance = 1000,
matrix_name = 'rw_beta',
reassemble = True,
cleanup = False)
tr.clone_df['B07'] = (tr.clone_df['hla-b'].str.startswith("B*07") | tr.clone_df['hla-b_1'].str.startswith("B*07"))
tr.clone_df['B07'] = ["B*07" if (x) else "NOTB*07 " for x in tr.clone_df['B07']]
#sparse.save_npz("S.npz", S)
from tcrdist.rep_funcs import compute_n_tally_out_of_memory
nn_tally_df_cohort = compute_n_tally_out_of_memory(fragments,
matrix_name = "rw_beta",
pm_processes = 6,
to_file = False,
to_memory = True,
knn_radius = 25,
x_cols = ['B07'])
from hierdiff.association_testing import cluster_association_test
nn_associations = cluster_association_test(res = nn_tally_df_cohort, y_col='cmember', method='fishers')
nn_associations = nn_associations.sort_values('pvalue', ascending = True)
import ast
nn_associations['neighbors_i'] = nn_associations.neighbors.apply(lambda x: ast.literal_eval(x))
from tcrdist.summarize import test_for_almost_subsets, filter_is, filter_gt
nn_associations['mostly_unique'] = test_for_almost_subsets(nn_associations['neighbors_i'], thr = 5)
nr_nn_associations = filter_is(nn_associations, 'mostly_unique', 1).copy()
#nr_nn_associations = filter_gt(nr_nn_associations, 'K_neighbors', 25).copy()
nr_nn_associations
# MOTIF GENERATION
from tcrsampler.sampler import TCRsampler
t = TCRsampler()
if 'olga_human_beta_t.sampler.tsv' not in t.currently_available_backgrounds():
t.download_background_file('olga_sampler.zip')
#t.download_background_file('olga_sampler.zip') # ONLY IF NOT ALREADY DONE
tcrsampler_beta = TCRsampler(default_background = 'olga_human_beta_t.sampler.tsv')
tcrsampler_beta.build_background(max_rows = 1000)
"""SEE PALMOTIF DOCS (https://github.com/agartland/palmotif)"""
from palmotif import compute_pal_motif, svg_logo
from tcrdist.summarize import _select
"""GENERATE SVG GRAPHIC FOR EACH NODE OF THE TREE"""
#pwmat_str = 'pw_beta'
cdr3_name = 'cdr3_b_aa'
gene_names = ['v_b_gene','j_b_gene']
svgs_beta = list()
svgs_beta_raw = list()
info_list = list()
from tcrdist.rep_diff import member_summ
summary = member_summ( res_df = nr_nn_associations,
clone_df = tr.clone_df,
addl_cols=['cohort','hla-a', 'hla-a_1', 'hla-b', 'hla-b_1', 'subject'])
nr_nn_associations = pd.concat([nr_nn_associations, summary], axis = 1).reset_index()
for i,r in nr_nn_associations.head(25).iterrows():
dfnode = tr.clone_df.iloc[r['neighbors_i'],:].copy()
# <pwnode> Pairwise Matrix for node sequences
pwnode = S[r['neighbors_i'],:] [:,r['neighbors_i']].todense()
if dfnode.shape[0] > 2:
iloc_idx = pwnode.sum(axis = 0).argmin()
centroid = dfnode[cdr3_name].to_list()[iloc_idx]
else:
centroid = dfnode[cdr3_name].to_list()[0]
print(f"CENTROID: {centroid}")
gene_usage_beta = dfnode.groupby(gene_names).size()
sampled_rep = tcrsampler_beta.sample( gene_usage_beta.reset_index().to_dict('split')['data'],
flatten = True, depth = max(100, 1000 // dfnode.shape[0]))
sampled_rep = [x for x in sampled_rep if x is not None]
motif, stat = compute_pal_motif(
seqs = _select(df = tr.clone_df,
iloc_rows = r['neighbors_i'],
col = cdr3_name),
refs = sampled_rep,
centroid = centroid)
svgs_beta.append(svg_logo(motif, return_str= True))
sampled_rep = sampled_rep.append(centroid)
motif_raw, _ = compute_pal_motif(
seqs =_select(df = tr.clone_df,
iloc_rows = r['neighbors_i'],
col = cdr3_name),
centroid = centroid)
svgs_beta_raw.append(svg_logo(motif_raw, return_str= True))
info_list.append(r)
def row_to_string(r, vals = ['ct_columns', 'val_0', 'ct_0', 'val_1', 'ct_1', 'val_2', 'ct_2','val_3', 'ct_3', 'levels', 'K_neighbors', 'R_radius', 'RR', 'OR', 'pvalue', 'FWERp','FDRq']):
#d = {v:r[v] for v in vals}
return "<br></br>".join([f"\t{v} : {r[v]}" for v in vals])
def to_html_table(r, vals = ['ct_columns', 'hla-a', 'hla-a_1', 'hla-b', 'hla-b_1', 'val_0', 'ct_0', 'val_2', 'ct_2', 'K_neighbors', 'R_radius', 'pvalue', 'FDRq','cdr3_b_aa','v_b_gene', 'j_b_gene', 'cohort','subject']):
return pd.DataFrame(r[vals]).transpose().to_html()
def shrink(html_str):
return html_str.replace('height="100%"', 'height="10%"').\
replace('width="100%"', 'width="10%"')
with open('svgs_in_line.html', 'w') as fh:
fh.write(f"<html><body>\n")
for svg, svg_raw, details in zip(svgs_beta, svgs_beta_raw, info_list):
fh.write(f"{shrink(svg_raw)}{shrink(svg)}")
try:
fh.write(to_html_table(details))
except:
print("F")
fh.write("<div></div>")
fh.write(f"</html></body>\n")
def make_motif_logo(tcrsampler,
clone_df,
pwmat,
centroid='CASSPDIEKYF',
v_gene='TRBV7-9*01',
radius=24,
pwmat_str='pw_delta',
cdr3_name='cdr3_d_aa',
v_name='v_d_gene',
gene_names=['v_d_gene', 'j_d_gene']):
"""
Make a motif from a tcrrep clone_df, pwmat, and a tcrsampler.
Parameters
----------
tcrsampler : tcrsamper.TCRsampler,
clone_df : pd.DataFrame,
pwmat : np.array,
centroid : str
e.g.,'CASSPDIEKYF',
v_gene : str
e.g. 'TRBV7-9*01',
radius = int
e.g., 26,
pwmat_str : str
e.g.,'pw_delta',
cdr3_name : str
e.g., 'cdr3_d_aa',
v_name : str
e.g., 'v_d_gene',
gene_names : list
eg., ['v_d_gene','j_d_gene']
Returns
-------
svg : str
svg_raw : str
Notes
-----
There is a safety first, efficiency loss involved
since we are relocating neighbors that
may already be know, but by looking
up the row index <irow> fisrst matching V,CDR3 this
function can be evoked without knowing
anything about the positions of
the neighbors ahead of time. This is particularly useful
since clone_df order is not stable after groupby
and deduplication.
"""
irow = clone_df[(clone_df[cdr3_name] == centroid)
& (clone_df[v_name] == v_gene)].index[0]
dfnode = clone_df[pd.Series(pwmat[irow, :]) <= radius].copy()
dfnode[gene_names[0]] = dfnode[gene_names[0]].apply(lambda x: allele_01(x))
dfnode[gene_names[1]] = dfnode[gene_names[1]].apply(lambda x: allele_01(x))
gene_usage = dfnode.groupby(gene_names).size()
sampled_rep = tcrsampler.sample(
gene_usage.reset_index().to_dict('split')['data'],
flatten=True,
depth=100)
sampled_rep = [x for x in sampled_rep if x is not None]
motif, stat = compute_pal_motif(seqs=dfnode[cdr3_name],
refs=sampled_rep,
centroid=centroid)
svg = svg_logo(motif, return_str=True)
motif_raw, _ = compute_pal_motif(seqs=dfnode[cdr3_name], centroid=centroid)
svg_raw = svg_logo(motif_raw, return_str=True)
return svg, svg_raw
iloc_idx = pwnode.sum(axis = 0).argmin()
centroid = dfnode[cdr3_name].to_list()[iloc_idx]
else:
centroid = dfnode[cdr3_name].to_list()[0]
print(f"ALPHA-CHAIN CENTROID: {centroid}")
gene_usage_alpha = dfnode.groupby(gene_names).size()
sampled_rep = tcrsampler_alpha.sample( gene_usage_alpha.reset_index().to_dict('split')['data'],
flatten = True, depth = 10)
sampled_rep = [x for x in sampled_rep if x is not None]
motif, stat = compute_pal_motif(
seqs = _select(df = tr.clone_df,
iloc_rows = r['neighbors_i'],
col = cdr3_name),
refs = sampled_rep,
centroid = centroid)
svgs_alpha.append(svg_logo(motif, return_str= True))
sampled_rep = sampled_rep.append(centroid)
motif_raw, _ = compute_pal_motif(
seqs =_select(df = tr.clone_df,
iloc_rows = r['neighbors_i'],
col = cdr3_name),
centroid = centroid)
svgs_alpha_raw.append(svg_logo(motif_raw, return_str= True))
"""Add Alpha SVG graphics to hcluster_df"""
tr.hcluster_df['svg_alpha'] = svgs_alpha
tr.hcluster_df['svg_alpha_raw'] = svgs_alpha_raw
def _tcrsampler_svgs(tcrrep,
default_background=None,
default_background_if_missing=None,
cdr3_name='cdr3_b_aa',
pwmat_str='pw_cdr3_b_aa',
chain='beta',
gene_names=['v_b_gene', 'j_b_gene'],
combine_olga=False,
verbose=True):
"""
Breath. What does this do?
Given a TCRrep instance, this function samples a background repertoire
using TCRsampler and makes svg-logos using palmotif.
This function doesn't return anything it. It needs to access
attribute values of a TCRrep (tcrrep) instance and
it modifies th etcrrep in place adding svgs and stats colums
to .hcluster_df_detailed DataFrame. TODO: could just output a dataframe
which would then just be concattenated.
ONLY WORKS WITH _BETA using defaults:
Notes
-----
Note: TCRSampler.build_background() accepts kwargs, we've set these as fixed as most user
won't know what these do and won't need to change them.
max_rows : int
Maximum clones per v,j pair (per subject)
stratify_by_subject : bool
If True, max_rows will apply to v,j,subject. If False, max_rows applies to v,j
use_frequency : bool
If True, uses frequency for ranking rows. If False, uses raw counts.
make_singleton : bool
If True, background is still sorted by frequency or counts,
but final fequency and counts values are overridden
and set to 1.
"""
from tcrsampler.sampler import TCRsampler
from palmotif import compute_pal_motif, svg_logo
import pandas as pd
from tcrdist.summarize import _select
if chain == 'alpha' and tcrrep.organism == "mouse":
# Here we enforce the rule that alpha-mouse cannot use an olga-sampler
# TODO: This should be removed as soon as TCRsampler can be updated with a valid
# mouse-alpha simulated background.
combine_olga = False
# _default_sampler returns a TCRSampler based on organism and chain
if verbose: print(f"INITIALIZING A TCRSAMPLER")
print(tcrrep.organism, chain)
t = _default_sampler(organism=tcrrep.organism, chain=chain)(
default_background=default_background,
default_background_if_missing=default_background_if_missing)
build_kargs = {
'max_rows': 100,
'stratify_by_subject': True,
'use_frequency': True,
'make_singleton': False
}
build_kargs_olga = {
'max_rows': 1000,
'stratify_by_subject': False,
'use_frequency': True,
'make_singleton': False
}
if verbose:
print(f"BUILDING A DEEPER BACKGROUND {report_kwargs(build_kargs)}")
t.build_background(**build_kargs)
# Olga Sampler
if combine_olga:
t_olga = _default_sampler_olga(chain=chain, organism=tcrrep.organism)()
if verbose:
print(
f"BUILDING A DEEPER BACKGROUND {report_kwargs(build_kargs_olga)}"
)
t.build_background(**build_kargs_olga)
olga_model = {
('beta', 'human'):
OlgaModel(recomb_type="VDJ", chain_folder="human_T_beta"),
('alpha', 'human'):
OlgaModel(recomb_type="VJ", chain_folder="human_T_alpha"),
('beta', 'mouse'):
OlgaModel(recomb_type="VDJ", chain_folder="mouse_T_beta")
}[(chain, tcrrep.organism)]
if 'prune' not in tcrrep.hcluster_df.columns:
if verbose: print("NO PRUNE COLUMNS USED ALL SET TO 0")
tcrrep.hcluster_df['prune'] = 0
print("ITERATE THROUGH CLUSTERS")
svgs = list()
svgs_raw = list()
reference_unique = list()
reference_unique_olga = list()
reference_size = list()
reference_size_olga = list()
percent_missing_sampler = list()
percent_missing_sampler_olga = list()
n_rows = tcrrep.hcluster_df.shape[0]
bar = IncrementalBar(f'Make {chain} SVGs :',
max=n_rows,
suffix='%(percent)d%%')
for i, r in tcrrep.hcluster_df.iterrows():
bar.next()
if r['prune'] == 0:
# <dfnode> is dataframe with all the clones at a given tree node
dfnode = tcrrep.clone_df.iloc[r['neighbors_i'], ].copy()
# <pwnode> Pairwise Matrix for node sequences
pwnode = getattr(
tcrrep,
pwmat_str)[r['neighbors_i'], :][:, r['neighbors_i']].copy()
iloc_idx = pwnode.sum(axis=0).argmin()
centroid = dfnode[cdr3_name].to_list()[iloc_idx]
# Compute gene usage at the node
# Convert to allele_01
for gene_name in gene_names:
dfnode[gene_name] = dfnode[gene_name].apply(
lambda x: allele_01(x))
gene_usage = dfnode.groupby(
gene_names).size() # e.g., ['v_b_gene','j_b_gene']
gene_usage_tuples = gene_usage.reset_index().to_dict(
'split')['data']
# Given gene usage use the <t> a TCRsampler instance to get background seqs
# Adjust depth for small nodes
adjust_depth = 10 * round(10 / dfnode.shape[0])
if adjust_depth < 10:
adjust_depth = 10
sampled_rep = t.sample(gene_usage_tuples,
flatten=True,
depth=adjust_depth * 10)
# Only keep the non-none sequences
sampled_rep = [x for x in sampled_rep if x is not None]
# < missing_gene > Count the percentage missing, sampler returns none when no v,j pair is present
expected_depth = dfnode.shape[0] * adjust_depth * 10
recovered_depth = len(sampled_rep)
percent_missing = round(
100 * (1 - (recovered_depth / expected_depth)), 1)
percent_missing_sampler.append(f"{percent_missing}%")
reference_unique.append(str(pd.Series(sampled_rep).nunique()))
reference_size.append(str(pd.Series(sampled_rep).count()))
if combine_olga:
# We modified Olga source code slightly, such that we simulated sequences
# with a given V,J gene usage
# OLD METHOD WHERE WE ACTUALLY SAMPLED, slower but can go much deeper. I don't think one rare sequence however, really make a big difference.
#flatten = lambda l: [item for sublist in l for item in sublist]
#sampled_rep_olga = [olga_model.gen_cdr3s(allele_01(v),allele_01(j),n*adjust_depth*10) for v,j,n in gene_usage_tuples]
#sampled_rep_olga = [x for x in flatten(sampled_rep_olga) if x is not None]
sampled_rep_olga = t_olga.sample(gene_usage_tuples,
flatten=True,
depth=adjust_depth * 10)
sampled_rep_olga = [
x for x in sampled_rep_olga if x is not None
]
expected_depth = dfnode.shape[0] * adjust_depth * 10
recovered_depth = len(sampled_rep_olga)
percent_missing_olga = round(
100 * (1 - (recovered_depth / expected_depth)), 1)
percent_missing_sampler_olga.append(f"{percent_missing_olga}%")
reference_unique_olga.append(
str(pd.Series(sampled_rep_olga).nunique()))
reference_size_olga.append(
str(pd.Series(sampled_rep_olga).count()))
# HERE WE COMBINE INTO A SINGLE BACKGROUND:
sampled_rep = sampled_rep + sampled_rep_olga
# Get motif matrix and motif stats
motif, stat = compute_pal_motif(seqs=_select(
df=tcrrep.clone_df, iloc_rows=r['neighbors_i'], col=cdr3_name),
refs=sampled_rep,
centroid=centroid)
svgs.append(svg_logo(motif, return_str=True))
# repeaat without references
raw_motif, raw_stat = compute_pal_motif(seqs=_select(
df=tcrrep.clone_df, iloc_rows=r['neighbors_i'], col=cdr3_name),
centroid=centroid)
# Convert the motif matrix into an svg_logo, append to list
svgs_raw.append(svg_logo(raw_motif, return_str=True))
else:
# If prune column is 1 don't go to the trouble of sampling and generating seqs
svgs.append("PRUNE")
svgs_raw.append("PRUNE")
reference_size.append("PRUNE")
reference_unique.append("PRUNE")
percent_missing_sampler.append("PRUNE")
percent_missing_sampler_olga.append("PRUNE")
reference_unique_olga.append("PRUNE")
reference_size_olga.append("PRUNE")
bar.next()
bar.finish()
# The standard svg_ includes background, whereas raw has no background
tcrrep.hcluster_df_detailed[f'svg_{chain}'] = svgs
tcrrep.hcluster_df_detailed[f'svg_raw_{chain}'] = svgs_raw
tcrrep.hcluster_df_detailed[f'ref_size_{chain}'] = reference_size
tcrrep.hcluster_df_detailed[f'ref_unique_{chain}'] = reference_unique
tcrrep.hcluster_df_detailed[
f'percent_missing_{chain}'] = percent_missing_sampler
if combine_olga:
tcrrep.hcluster_df_detailed[
f'ref_size_olga_{chain}'] = reference_size_olga
tcrrep.hcluster_df_detailed[
f'ref_unique_olga_{chain}'] = reference_unique_olga
tcrrep.hcluster_df_detailed[
f'percent_missing_olga_{chain}'] = percent_missing_sampler_olga
return True