FiRE - Finder of Rare Entities

Update: FiRE is now available via cran. install FiRE using

install.packages('FiRE')

Introduction
External dependencies
Installation
Python Package
-Prerequisites
-Installation Steps
-Usage
R Package
-Prerequisites
-Installation Steps
-Usage
Publication
Copyright
Patent

Introduction

Tested on Ubuntu 14.04 and Ubuntu 16.04.

All results in manuscript have been generated using python module.

FiRE is available for python and R. Required versions and modules for both are as mentioned below. cpp modules are necessary for both of them.

External Dependencies

Following packages are required to run/install the FiRE software.

Required cpp modules

    g++ >= 4.8.4
    boost >= 1.54.0

FiRE only needs <boost/random.hpp> from boost. So, full installation is not necessary. It can be downloaded from boost.org and used as is.

Installation

    [sudo] ./INSTALL [ --boost-path <boost-path> | --log-file <log-file> | --inplace | --py | --R | --help ]
    [sudo] ./UNINSTALL_python
    [sudo] ./UNINSTALL_R

    --boost-path <boost-path>  : python        : Path to boost-library, if boost is not installed at default location, this value needs to be provided.
    --inplace                  : python        : Required only for python, if set, inplace build will be run and resulting lib will be stored in python/FiRE.
    --log-file <log-file>      : python        : Required only for python, ignored with --inplace set.
    --py                       : python        : Install FiRE in python environment.
    --R                        : R             : Install FiRE in R environment.
    --help                     : python | R    : Display this help.

    Info:

    UNINSTALL_[python | R] files are generated upon installation.

Typically, FiRE module takes a few seconds to install. A snippet of installation time taken by FiRE (in seconds) on a machine with Intel® Core™ i5-7200U (CPU @ 2.50GHz × 4), with 8GB memory, and OS Ubuntu 16.04 LTS is as follows

real 2.92
user 2.73
sys 0.18

Python Package

Prerequisites

Required python modules

    python 2.7
    # [EDIT] python 3 can also be used with standard installation. 
    # however, with --inplace option uninstallation may fail. 
    # As a workround generated .so file can be removed manually if uninstallation of FiRE is desired.

For FiRE module

    cython >= 0.23.4
    distutils >= 2.7.12

For preprocessing

    numpy >= 1.13.3
    pandas >= 0.20.3
    statsmodels >= 0.8.0

For demo

    gzip >= 1.2.11 (zlib)
    scipy >= 1.1.0
    matplotlib >= 2.1.0
    cmocean >= 1.2
    sklearn >= 0.19.1

Installation Steps

with virtual environment avoid using sudo. (Thanks to chenxofhit)

[sudo] chomd +x ./INSTALL

If boost is installed at default location

[sudo] ./INSTALL --py

If boost is installed at custom location

[sudo] ./INSTALL --boost-path <full-path> --py

Example:

[sudo] ./INSTALL --boost-path $HOME/boost/boost_1_54_0 --py

Above installation steps will generate fireInstall.log file. It is advisable to keep this file, since it will be needed for uninstallation. Name of the log file can be modified during installation.

./INSTALL --log-file <log-file-name> --py

Above steps will install FiRE at the default location.

For inplace installation

./INSTALL --inplace --py

Uninstallation of FiRE Software.

[sudo] ./UNINSTALL_python

Usage

Run demo from FiRE directory as follows

python example/jurkat_simulation.py

Since data (data/jurkat_two_species_1580.txt.gz) is large, this may require large amount of RAM to load and pre-process. We have also providee pre-processed data (data/preprocessedData_jurkat_two_species_1580.txt.gz). Pre-processing was done using the script present in utils/preprocess.py. Demo using this data as follows

python example/jurkat_simulation_small.py

Small demo takes seconds to complete. Exact time taken by the demo on a machine with Intel® Core™ i5-7200U (CPU @ 2.50GHz × 4), with 8GB memory, and OS Ubuntu 16.04 LTS is as follows

Loading preprocessed Data : 1.850723s
Running FiRE : 1.134673s

Total Demo time:

real 4.33
user 3.55
sys 0.76

Step-by-step description of full demo (example/jurkat_simulation.py) is as follows

Load libraries

```python import sys sys.path.append(‘utils’)

import numpy as np import gzip from scipy import stats

import preprocess as pp import misc import FiRE

2. <h4>Load Data in current environment.</h4>
```python
#Data matrix should only consist of values where rows represent cells and columns represent genes.

with gzip.GzipFile('data/jurkat_two_species_1580.txt.gz', 'r') as fid:
    data = np.genfromtxt(fid)

data = data.T #Samples * Features

labels = np.genfromtxt('data/labels_jurkat_two_species_1580.txt', dtype=np.int) #Cells with label '1' represent abundant, while cells with label '2' represent rare.

Data Pre-processing

Call function ranger_preprocess for selecting thousand variable genes.

```python

#Genes genes = np.arange(1, data.shape[1]+1) #It can be replaced with original gene names

#Filter top 1k genes preprocessedData, selGenes = pp.ranger_preprocess(data, genes, optionToSave=True, dataSave=outputFolder)

|Parameter | Description | Required or Optional| Datatype | Default Value |
| -----:| -----:| -----:|-----:|-----:|
|data | Data for processing | Required | `np.array [nCells, nGenes]` | - |
|genes | Names of Genes | Required | `np.array [nGenes]` | - |
|ngenes_keep | Number of genes to keep | Optional | `integer` | 1000 |
|dataSave | Path to save results | Optional | `string` | Current working Directory (Used only when optionToSave is True) |
|optionToSave | Save processed output or not | Optional | `boolean` | False(Does not save) |
|minLibSize | Minimum number of expressed features | Optional | `integer` | 0 |
|verbose | Display progress | Optional | `boolean` | True(Prints intermediate results) |

```python
'''
Returned Value :
    preprocessedData : processed data matrix (log2 transformed) : np.array [nCells, nVariableGenes]
    selGenes         : Names of thousand variable genes selected : np.array [nVariableGenes]
'''

Create model of FiRE.

model = FiRE.FiRE(L=100, M=50, H=1017881, seed=5489, verbose=0)

Parameter	Description	Required or Optional	Datatype	Default Value
L	Total number of estimators	Required	`int`	-
M	Number of features to be randomly sampled for each estimator	Required	`int`	-
H	Number of bins in hash table	Optional	`int`	1017881
seed	Seed for random number generator	Optional	`unsigned int`	5489
verbose	Controls verbosity of program at run time (0/1)	Optional	`int`	0 (silent)

Apply model to the above dataset.
```
model.fit(preprocessedData)
```
Calculate FiRE score of every cell.

```python score = np.array(model.score(preprocessedData)) ‘’’ Returned Value : score : FiRE score of every cell : np.array[nCells]

Higher values of FiRE score represent rare cells. ‘’’

7. <h4>Select cells with higher values of FiRE score, that satisfy IQR-based thresholding criteria.</h4>

```python

q3 = np.percentile(score, 75)
iqr = stats.iqr(score)
th = q3 + 1.5*iqr

indIqr = np.where(score >= th)[0]

dataSel = preprocessedData[indIqr,:] #Select subset of rare cells

#Create a file with binary predictions
predictions = np.zeros(data.shape[0])
predictions[indIqr] = 1 #Replace predictions for rare cells with '1'.

Access to model parameters.

Sampled dimensions can be accessed via
```
# type : 2d list
# shape : L x M
model.dims
```
Chosen thresholds can be accessed via
```
# type : 2d list
# shape : L x M
model.thresholds
```

Weights can be accessed via

# type : 2d list
# shape : L X M
model.weights

Hash tables can be accessed via

# type : 3d list
# shape : L x H x <dynamic>
# <dynamic> : as per number of samples in a bin (H) for a given estimator (L).
model.bins

FiRE recovers artifitially planted rare cells (Figure).

(a) t-SNE based 2D embedding of the cells with color-coded identities (b) FiRE score intensities plotted on the t-SNE based 2D map. (c) Rare cells detected by FiRE.

R Package

Prerequisites

Required R modules

    R >= 3.2.0

For FiRE module

    Rcpp >= 0.12.19
    BH >= 1.66

For preprocessing and demo

    Matrix >= 1.2.14
    plyr >= 1.8.4

Installation Steps

[sudo] chomd +x ./INSTALL

Installation of FiRE Software.

[sudo] ./INSTALL --R

Uninstallation of FiRE Software.

[sudo] ./UNINSTALL_R

Usage

Run demo from FiRE directory as follows

Rscript example/jurkat_simulation.R

Rscript example/jurkat_simulation_small.R

Small demo takes seconds to complete. Exact time taken by the demo on a machine with Intel® Core™ i5-7200U (CPU @ 2.50GHz × 4), with 8GB memory, and OS Ubuntu 16.04 LTS is as follows

Total Demo time:

real 4.11
user 3.16
sys 1.13

Step-by-step description of full demo (example/jurkat_simulation.R) is as follows

Load libraries

library('FiRE')
source('utils/preprocess.R')

Load Data in current environment.

```R #Read data data <- read.table(gzfile(‘data/jurkat_two_species_1580.txt.gz’)) data <- t(data) #Samples * Features

#Read Labels labels <- read.table(‘data/labels_jurkat_two_species_1580.txt’) #Cells with label ‘1’ represent abundant, while cells with label ‘2’ represent rare.

#Genes genes <- c(1:dim(data)[2]) #It can be replaced with original gene names

data_mat <- list(mat=data, gene_symbols=genes)

3. <h4>Call function ranger_preprocess for selecting thousand variable genes.</h4>
```R
preprocessedList <- ranger_preprocess(data_mat)
preprocessedData <- as.matrix(preprocessedList$preprocessedData)

Parameter	Description	Required or Optional	Datatype	Default Value
data_mat	List consisting of data for processing and gene symbols	Required	`list(mat=data, gene_symbols=genes)`	-
ngenes_keep	Number of genes to keep	Optional	`integer`	1000
dataSave	Path to save results	Optional	`string`	Current working Directory (Used only when optionToSave is True)
optionToSave	Save processed output or not	Optional	`boolean`	False(Does not save)
minLibSize	Minimum number of expressed features	Optional	`integer`	0
verbose	Display progress	Optional	`boolean`	True(Prints intermediate results)

Create model of FiRE.

# model <- new(FiRE::FiRE, L, M, H, seed, verbose)
model <- new(FiRE::FiRE, 100, 50, 1017881, 5489, 0)

Parameter	Description	Required or Optional	Datatype	Default Value
L	Total number of estimators	Required	`int`	-
M	Number of features to be randomly sampled for each estimator	Required	`int`	-
H	Number of bins in hash table	Optional	`int`	1017881
seed	Seed for random number generator	Optional	`int`	5489
verbose	Controls verbosity of program at run time (0/1)	Optional	`int`	0 (silent)

Apply model to the above dataset.
```
model$fit(preprocessedData)
```
Acceptable datatype is of matrix class and of type double (Numeric matrix).

Calculate FiRE score of every cell.

# Returns a numeric vector
score <- model$score(preprocessedData)

Select cells with higher values of FiRE score, that satisfy IQR-based thresholding criteria.

```R #Apply IQR-based criteria to identify rare cells for further downstream analysis. q3 <- quantile(score, 0.75) iqr <- IQR(score) th <- q3 + (1.5*iqr)

#Select indexes that satisfy IQR-based thresholding criteria. indIqr <- which(score >= th)

#Create a file with binary predictions predictions <- integer(dim(data)[1]) predictions[indIqr] <- 1 #Replace predictions for rare cells with ‘1’.

8. <h4>Access to model parameters.</h4>
Sampled dimensions can be accessed via
```R
# type : Integer matrix
# shape : L x M
model$d

Chosen thresholds can be accessed via

# type : Numeric matrix
# shape : L x M
model$ths

Weights can be accessed via

# type : Numeric matrix
# shape : 0 x 0
model$w

Hash tables can be accessed via

# type : List
# shape : L x H x <dynamic>
# <dynamic> : as per number of samples in a bin (H) for a given estimator (L).
model$b

Publication

Jindal, A., Gupta, P., Jayadeva and Sengupta, D., 2018. Discovery of rare cells from voluminous single cell expression data. Nature communications, 9(1), p.4719. DOI: https://doi.org/10.1038/s41467-018-07234-6

Copyright

This software package is distributed under GNU GPL v3.

This work is free to use for academic and research purposes. Please contact Dr. Debarka for commercial use of this work.

FiRE - Finder of Rare Entities

Contents

Introduction

External Dependencies

Required cpp modules

Installation

Python Package

Prerequisites

Required python modules

Installation Steps

If boost is installed at default location

If boost is installed at custom location

For inplace installation

Uninstallation of FiRE Software.

Usage

Load libraries

Data Pre-processing

Call function ranger_preprocess for selecting thousand variable genes.

Create model of FiRE.

Apply model to the above dataset.

Calculate FiRE score of every cell.

Access to model parameters.

FiRE recovers artifitially planted rare cells (Figure).

R Package

Prerequisites

Required R modules

Installation Steps

Installation of FiRE Software.

Uninstallation of FiRE Software.

Usage

Load libraries

Load Data in current environment.

Create model of FiRE.

Apply model to the above dataset.

Calculate FiRE score of every cell.

Select cells with higher values of FiRE score, that satisfy IQR-based thresholding criteria.

Publication

Copyright