SIAMCAT input files formats
Date last modified: 2018-09-24
Contents
1 Introduction
This vignette illustrates how to read and input your own data to the
SIAMCAT package. We will cover reading in text files from the disk,
formatting them and using them to create an object of siamcat-class.
The siamcat-class is the centerpiece of the package. All of the input data
and result are stored inside of it. The structure of the object is described
below in the siamcat-class object section.
2 Loading your data into R
2.1 SIAMCAT input
Generally, there are three types of input for SIAMCAT:
2.1.1 Features
The features should be a matrix, a data.frame, or an otu_table,
organized as follows:
features (in rows) x samples (in columns).
| Sample_1 | Sample_2 | Sample_3 | Sample_4 | Sample_5 | |
|---|---|---|---|---|---|
| Feature_1 | 0.59 | 0.71 | 0.78 | 0.61 | 0.66 |
| Feature_2 | 0.00 | 0.02 | 0.00 | 0.00 | 0.00 |
| Feature_3 | 0.02 | 0.00 | 0.00 | 0.00 | 0.20 |
| Feature_4 | 0.34 | 0.00 | 0.13 | 0.07 | 0.00 |
| Feature_5 | 0.06 | 0.16 | 0.00 | 0.00 | 0.00 |
Please note that
SIAMCATis supposed to work with relative abundances. Other types of data (e.g. counts) will also work, but not all functions of the package will result in meaningful outputs.
An example of a typical feature file is attached to the SIAMCAT package,
containing data from a publication investigating the microbiome in colorectal
cancer (CRC) patients and controls (the study can be found here:
Zeller et al). The metagenomics
data were processed with the MOCAT pipeline, returning
taxonomic profiles on the species levels (specI):
library(SIAMCAT)
fn.in.feat <- system.file(
"extdata",
"feat_crc_zeller_msb_mocat_specI.tsv",
package = "SIAMCAT"
)One way to load such data into R could be the use of read.table
(Beware of the defaults in R! They are not always useful…)
feat <- read.table(fn.in.feat, sep='\t',
header=TRUE, quote='',
stringsAsFactors = FALSE, check.names = FALSE)
# look at some features
feat[110:114, 1:2]## CCIS27304052ST-3-0 CCIS15794887ST-4-0
## Bacteroides caccae [h:1096] 1.557937e-03 1.761949e-03
## Bacteroides eggerthii [h:1097] 2.734527e-05 4.146882e-05
## Bacteroides stercoris [h:1098] 1.173786e-03 2.475838e-03
## Bacteroides clarus [h:1099] 4.830533e-04 4.589747e-06
## Methanohalophilus mahii [h:11] 0.000000e+00 0.000000e+002.1.2 Metadata
The metadata should be either a matrix or a data.frame.
samples (in rows) x metadata (in columns):
| Age | Gender | BMI | |
|---|---|---|---|
| Sample_1 | 52 | 1 | 20 |
| Sample_2 | 37 | 1 | 18 |
| Sample_3 | 66 | 2 | 24 |
| Sample_4 | 54 | 2 | 26 |
| Sample_5 | 65 | 2 | 30 |
The rownames of the metadata should match the colnames of the feature
matrix.
Again, an example of such a file is attached to the SIAMCAT package, taken
from the same study:
fn.in.meta <- system.file(
"extdata",
"num_metadata_crc_zeller_msb_mocat_specI.tsv",
package = "SIAMCAT"
)Also here, the read.table can be used to load the data into R.
meta <- read.table(fn.in.meta, sep='\t',
header=TRUE, quote='',
stringsAsFactors = FALSE, check.names = FALSE)
head(meta)## age gender bmi diagnosis localization crc_stage fobt
## CCIS27304052ST-3-0 52 1 20 0 NA 0 0
## CCIS15794887ST-4-0 37 1 18 0 NA 0 0
## CCIS74726977ST-3-0 66 2 24 1 NA 0 0
## CCIS16561622ST-4-0 54 2 26 0 NA 0 0
## CCIS79210440ST-3-0 65 2 30 0 NA 0 1
## CCIS82507866ST-3-0 57 2 24 0 NA 0 0
## wif_test
## CCIS27304052ST-3-0 0
## CCIS15794887ST-4-0 0
## CCIS74726977ST-3-0 NA
## CCIS16561622ST-4-0 0
## CCIS79210440ST-3-0 0
## CCIS82507866ST-3-0 02.1.3 Label
Finally, the label can come in different three different flavours:
Named vector: A named vector containing information about cases and controls. The names of the vector should match the
rownamesof the metadata and thecolnamesof the feature data. The label can contain either the information about cases and controls either- as integers (e.g.
0and1), - as characters (e.g.
CTRandIBD), or - as factors.
- as integers (e.g.
Metadata column: You can provide the name of a column in the metadata for the creation of the label. See below for an example.
Label file:
SIAMCAThas a function calledread.label, which will create a label object from a label file. The file should be organized as follows:- The first line is supposed to read:
#BINARY:1=[label for cases];-1=[label for controls] - The second row should contain the sample identifiers as tab-separated list (consistent with feature and metadata).
- The third row is then supposed to contain the actual class labels
(tab-separated):
1for each case and-1for each control.
An example file is attached to the package again, if you want to have a look at it.
- The first line is supposed to read:
For our example dataset, we can create the label object out of the metadata
column called diagnosis:
label <- create.label(meta=meta, label="diagnosis",
case = 1, control=0)When we later plot the results, it might be nicer to have names for the
different groups stored in the label object (instead of 1 and 0). We can
also supply them to the create.label function:
label <- create.label(meta=meta, label="diagnosis",
case = 1, control=0,
p.lab = 'cancer', n.lab = 'healthy')## Label used as case:
## 1
## Label used as control:
## 0## + finished create.label.from.metadata in 0.001 slabel$info## healthy cancer
## -1 1Note:
If you have no label information for your dataset, you can still create aSIAMCATobject from your features alone. TheSIAMCATobject without label information will contain aTESTlabel that can be used for making holdout predictions. Other functions, e.g. model training, will not work on such an object.
2.2 LEfSe format files
LEfSe is a tool for identification of associations between micriobial features and up to two metadata. LEfSe uses LDA (linear discriminant analysis).
LEfSe input file is a .tsv file. The first few rows contain the metadata. The
following row contains sample names and the rest of the rows are occupied by
features. The first column holds the row names:
| label | healthy | healthy | healthy | cancer | cancer |
|---|---|---|---|---|---|
| age | 52 | 37 | 66 | 54 | 65 |
| gender | 1 | 1 | 2 | 2 | 2 |
| Sample_info | Sample_1 | Sample_2 | Sample_3 | Sample_4 | Sample_5 |
| Feature_1 | 0.59 | 0.71 | 0.78 | 0.61 | 0.66 |
| Feature_2 | 0.00 | 0.02 | 0.00 | 0.00 | 0.00 |
| Feature_3 | 0.02 | 0.00 | 0.00 | 0.00 | 0.00 |
| Feature_4 | 0.34 | 0.00 | 0.43 | 0.00 | 0.00 |
| Feature_5 | 0.56 | 0.56 | 0.00 | 0.00 | 0.00 |
An example of such a file is attached to the SIAMCAT package:
fn.in.lefse<- system.file(
"extdata",
"LEfSe_crc_zeller_msb_mocat_specI.tsv",
package = "SIAMCAT"
)SIAMCAT has a dedicated function to read LEfSe format files. The read.lefse
function will read in the input file and extract metadata and features:
meta.and.features <- read.lefse(fn.in.lefse,
rows.meta = 1:6, row.samples = 7)
meta <- meta.and.features$meta
feat <- meta.and.features$featWe can then create a label object from one of the columns of the meta object and
create a siamcat object:
label <- create.label(meta=meta, label="label", case = "cancer")## Label used as case:
## cancer
## Label used as control:
## healthy## + finished create.label.from.metadata in 0.002 s2.3 metagenomeSeq format files
metagenomeSeq is an R package to determine differentially abundant features between multiple samples.
There are two ways to input data into metagenomeSeq:
- two files, one for metadata and one for features - those can be used
in
SIAMCATjust like described in SIAMCAT input withread.table:
fn.in.feat <- system.file(
"extdata",
"CHK_NAME.otus.count.csv",
package = "metagenomeSeq"
)
feat <- read.table(fn.in.feat, sep='\t',
header=TRUE, quote='', row.names = 1,
stringsAsFactors = FALSE, check.names = FALSE
)BIOMformat file, that can be used inSIAMCATas described in the following section
2.4 BIOM format files
The BIOM format files can be added to SIAMCAT via phyloseq. First the file
should be imported using the phyloseq function import_biom. Then a
phyloseq object can be imported as a siamcat object as descibed in the
next section.
2.5 Creating a siamcat object of a phyloseq object
The siamcat object extends on the phyloseq object. Therefore, creating
a siamcat object from a phyloseq object is really straightforward. This
can be done with the siamcat constructor function. First, however, we need
to create a label object:
data("GlobalPatterns") ## phyloseq example data
label <- create.label(meta=sample_data(GlobalPatterns),
label = "SampleType",
case = c("Freshwater", "Freshwater (creek)", "Ocean"))## Label used as case:
## Freshwater,Freshwater (creek),Ocean
## Label used as control:
## rest## + finished create.label.from.metadata in 0.003 s# run the constructor function
siamcat <- siamcat(phyloseq=GlobalPatterns, label=label)## + starting validate.data## +++ checking overlap between labels and features## + Keeping labels of 26 sample(s).## +++ checking sample number per class## Data set has a limited number of training examples:
## rest 18
## Case 8
## Note that a dataset this small/skewed is not necessarily suitable for analysis in this pipeline.## +++ checking overlap between samples and metadata## + finished validate.data in 0.151 s3 Creating a siamcat-class object
The siamcat-class is the centerpiece of the package. All of the is stored
inside of the object:
In the figure above, rectangles depict slots of the object and the class of
the object stored in the slot is given in the ovals. There are two
obligatory slots -phyloseq (containing the metadata as sample_data and
the original features as otu_table) and label - marked with thick borders.
The siamcat object is constructed using the siamcat() function. There are
two ways to initialize it:
Features: You can provide a feature
matrix,data.frame, orotu_tableto the function (together with label and metadata information):siamcat <- siamcat(feat=feat, label=label, meta=meta)phyloseq: The alternative is to create a
siamcatobject directly out of aphyloseqobject:siamcat <- siamcat(phyloseq=phyloseq, label=label)
Please note that you have to provide either feat or phyloseq and that
you cannot provide both.
In order to explain the siamcat object better we will show how each of the
slots is filled.
3.1 phyloseq, label and orig_feat slots
The phyloseq and label slots are obligatory.
- The phyloseq slot is an object of class
phyloseq, which is described in the help file of thephyloseqclass. Help can be accessed by typing into R console:help('phyloseq-class').- The
otu_tableslot inphyloseq-seehelp('otu_table-class')- stores the original feature table. ForSIAMCAT, this slot can be accessed byorig_feat.
- The
- The label slot contains a list. This list has a specific set of entries
-see
help('label-class')- that are automatically generated by theread.labelorcreate.labelfunctions.
The phyloseq, label and orig_feat are filled when the siamcat object is
first created with the constructor function.
3.2 All the other slots
Other slots are filled during the run of the SIAMCAT workflow:
3.3 Accessing and assigning slots
Each slot in siamcat can be accessed by typing
slot_name(siamcat)e.g. for the eval_data slot you can types
eval_data(siamcat)There is one notable exception: the phyloseq slot has to be accessed with
physeq(siamcat) due to technical reasons.
Slots will be filled during the SIAMCAT workflow by the package’s functions.
However, if for any reason a slot needs to be assigned outside of the workflow,
the following formula can be used:
slot_name(siamcat) <- object_to_assigne.g. to assign a new_label object to the label slot:
label(siamcat) <- new_labelPlease note that this may lead to unforeseen consequences…
3.4 Slots inside the slots
There are two slots that have slots inside of them. First, the model_list
slot has a models slot that contains the actual list of
mlr models
-can be accessed via models(siamcat)- and model.type which is a character
with the name of the method used to train the model: model_type(siamcat).
The phyloseq slot has a complex structure. However, unless the phyloseq
object is created outside of the SIAMCAT workflow, only two slots of phyloseq
slot will be occupied: the otu_table slot containing the features table and
the sam_data slot containing metadata information. Both can be accessed by
typing either features(siamcat) or meta(siamcat).
Additional slots inside the phyloseq slots do not have dedicated accessors,
but can easily be reached once the phyloseq object is exported from the
siamcat object:
phyloseq <- physeq(siamcat)
tax_tab <- tax_table(phyloseq)
head(tax_tab)## Taxonomy Table: [6 taxa by 7 taxonomic ranks]:
## Kingdom Phylum Class Order Family
## 549322 "Archaea" "Crenarchaeota" "Thermoprotei" NA NA
## 522457 "Archaea" "Crenarchaeota" "Thermoprotei" NA NA
## 951 "Archaea" "Crenarchaeota" "Thermoprotei" "Sulfolobales" "Sulfolobaceae"
## 244423 "Archaea" "Crenarchaeota" "Sd-NA" NA NA
## 586076 "Archaea" "Crenarchaeota" "Sd-NA" NA NA
## 246140 "Archaea" "Crenarchaeota" "Sd-NA" NA NA
## Genus Species
## 549322 NA NA
## 522457 NA NA
## 951 "Sulfolobus" "Sulfolobusacidocaldarius"
## 244423 NA NA
## 586076 NA NA
## 246140 NA NAIf you want to find out more about the phyloseq data structure, head over to the phyloseq BioConductor page. # Session Info
sessionInfo()## R version 4.4.0 beta (2024-04-15 r86425)
## Platform: x86_64-pc-linux-gnu
## Running under: Ubuntu 22.04.4 LTS
##
## Matrix products: default
## BLAS: /home/biocbuild/bbs-3.19-bioc/R/lib/libRblas.so
## LAPACK: /usr/lib/x86_64-linux-gnu/lapack/liblapack.so.3.10.0
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##
## attached base packages:
## [1] stats graphics grDevices utils datasets methods base
##
## other attached packages:
## [1] ggpubr_0.6.0 SIAMCAT_2.8.0 phyloseq_1.48.0 mlr3_0.19.0
## [5] lubridate_1.9.3 forcats_1.0.0 stringr_1.5.1 dplyr_1.1.4
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