ReSDK Tables
ReSDK tables are helper classes for aggregating collection data in tabular format. Currently, we have four flavours:
RNATables
Imagine you are modelling gene expression data from a given collection.
Ideally, you would want all expression values organized in a table where
rows represents samples and columns represent genes. Class
RNATables
gives you just that (and more).
We will present the functionality of RNATables
through an
example. We will:
Create an instance of
RNATables
and examine it’s attributesFetch raw expressions and select TIS signature genes with sufficient coverage
Normalize expression values (log-transform) and visualize samples in a simple PCA plot
First, connect to a Resolwe server, pick a collection and create
and instance of RNATables
:
import resdk
from resdk.tables import RNATables
res = resdk.Resolwe(url='https://app.genialis.com/')
res.login()
collection = res.collection.get("sum149-fresh-for-rename")
sum149 = RNATables(collection)
Object sum149
is an instance of RNATables
and has many attributes. For a complete list see
the SDK Reference, here we list the most commonly used ones:
# Expressions raw counts
sum149.rc
# Expressions normalized counts
sum149.exp
# See normalization method
sum149.exp.attrs["exp_type"]
# Sample metadata
sum149.meta
# Sample QC metrics
sum149.qc
# Dictionary that maps gene ID's into gene symbols
sum149.readable_columns
# This is handy to rename column names (gene ID's) to gene symbols
sum149.rc.rename(columns=sum149.readable_columns)
Note
Expressions and metadata are cached in memory as well as on disk. At each time they are re-requested a check is made that local and server side of data is synced. If so, cached data is used. Otherwise, new data will be pulled from server.
In our example we will only work with a set of TIS signature genes:
TIS_GENES = ["CD3D", "IDO1", "CIITA", "CD3E", "CCL5", "GZMK", "CD2", "HLA-DRA", "CXCL13", "IL2RG", "NKG7", "HLA-E", "CXCR6", "LAG3", "TAGAP", "CXCL10", "STAT1", "GZMB"]
We will identify low expressed genes and only keep the ones with average raw expression above 20:
tis_rc = sum149.rc.rename(columns=sum149.readable_columns)[TIS_GENES]
mean = tis_rc.mean(axis=0)
high_expressed_genes = mean.loc[mean > 20].index
Now, lets select TPM normalized expressions and keep only highly
expressed tis genes. We also transform to log2(TPM + 1)
:
import numpy as np
tis_tpm = sum149.exp.rename(columns=sum149.readable_columns)[high_expressed_genes]
tis_tpm_log = np.log(tis_tpm + 1)
Finally, we perform PCA and visualize the results:
from sklearn.decomposition import PCA
pca = PCA(n_components=2, whiten=True)
Y = pca.fit_transform(tis_tpm_log)
import matplotlib.pyplot as plt
for ((x, y), sample_name) in zip(Y, tis_tpm.index):
plt.plot(x, y, 'bo')
plt.text(x, y, sample_name)
plt.xlabel(f"PC1 ({pca.explained_variance_ratio_[0]})")
plt.ylabel(f"PC2 ({pca.explained_variance_ratio_[1]})")
plt.show()
MethylationTables
Similar as RNATables
provide access to raw counts and normalized
expression values of RNA data, MethylationTables
allow for fast
access of beta and m-values of methylation data:
meth = resdk.tables.MethylationTables(<collection-with-methylation-data>)
# Methylation beta-values
meth.beta
# Methylation m-values
meth.mval
MATables
Similar as RNATables
provide access to raw counts and normalized
expression values of RNA data, MATables
allow for fast
access of expression values per probe of microarray:
ma = resdk.tables.MATables(<collection-with-microarray-data>)
# Microarray expressions values (columns are probe ID's)
ma.exp
VariantTables
Similar as RNATables
provide access to raw counts and normalized
expression values of RNA data, VariantTables
allow for fast
access of variant data present in Data of type data:mutationstable
:
vt = resdk.tables.VariantTables(<collection-with-variant-data>)
vt.variants
The output of the above would look something like this:
sample_id |
chr1_123_C>T |
chr1_126_T>C |
---|---|---|
101 |
2 |
NaN |
102 |
0 |
2 |
In rows, there are sample ID’s. In columns there are variants where each
variant is given as:
<chromosome>_<position>_<nucleotide-change>
.
Values in table can be:
0 (wild-type / no mutation)
1 (heterozygous mutation),
2 (homozygous mutation)
NaN (QC filters are failing - mutation status is unreliable)
Inspecting depth
The reason for NaN values may be that the read depth on certain position is too low for GATK to reliably call a variant. In such case, it is worth inspecting the depth or depth per base:
# Similar as above but one gets depth on particular variant / sample
vt.depth
# One can also get depth for specific base
vt.depth_a
vt.depth_c
vt.depth_t
vt.depth_g
Filtering mutations
Process mutations-table
on Genialis Platform accepts either mutations
or
geneset
input which specifies the genes of interest. It restricts the scope
of mutation search to just a few given genes.
However, it can happen that not all the samples have the same mutations
or
geneset
input. In such cases, it makes little sense to merge the information
about mutations from multiple samples. By default, VariantTables
checks that
all Data is computed with same mutations
/ geneset
input. If this is
not true, it will raise an error.
But if you provide additional argument geneset
it will limit the
mutations to only those in the given geneset. An example:
# Sample 101 has mutations input "FHIT, BRCA2"
# Sample 102 has mutations input "BRCA2"
# This would cause error, since the mutations inputs are not the same
vt = resdk.tables.VariantTables(<collection>)
vt.variants
# This would limit the variants to just the ones in BRCA2 gene.
vt = resdk.tables.VariantTables(<collection>, geneset=["BRCA2"])
vt.variants