# escape
#### Easy single cell analysis platform for enrichment
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<img align="right" src="https://github.com/ncborcherding/escape/blob/dev/www/escape_hex.png" width="352" height="352">
### Introduction
Single-cell sequencing (SCS) is a fundamental technology in investigating a diverse array of biological fields. Part of the struggle with the high-resolution approach of SCS, is distilling the data down to meaningful scientific hypotheses. *escape* was created to bridge SCS results, either from raw counts or from popular R-based single-cell pipelines, like [Seurat](https://satijalab.org/seurat/) or [SingleCellExperiment](https://bioconductor.org/books/release/OSCA/book-contents.html#basics), with gene set enrichment analyses (GSEA). The *escape* package allows users to easily incorporate multiple methods of GSEA and offers several visualization and analysis methods. The package accesses the entire [Molecular Signature Database v7.0](https://www.gsea-msigdb.org/gsea/msigdb/search.jsp) and enables users to select single, multiple gene sets, and even libraries to perform enrichment analysis on.
#### Methods of GSEA Available
* Gene Set Variation Analysis (GSVA) - [citation](https://pubmed.ncbi.nlm.nih.gov/23323831/)
* Single-sample Gene Set Enrichment Analysis (ssGSEA) - [citation](https://pubmed.ncbi.nlm.nih.gov/19847166/)
* AUCell - [citation](https://pubmed.ncbi.nlm.nih.gov/28991892/)
* UCell -[citation](https://pubmed.ncbi.nlm.nih.gov/34285779/)
More information on each method is available in the *escape* manual for ```escape.matrix()``` and the citation links. If using these methods, users should cite the original works as well.
### Installation
#### GSVA requirement
*escape* requires GSVA v1.51.5 (not on Bioconductor 3.18). The easiest way to install is:
```r
devtools::install_github("rcastelo/GSVA")
```
#### Install Via GitHub
```r
devtools::install_github("ncborcherding/escape")
```
#### Install via Bioconductor
For now, the newest version of escape is available in the Bioconductor dev version (3.19).
```r
BiocManager::install(version='devel')
BiocManager::install("escape")
```
### Learning To Use escape:
#### Basic Usage
#### Running escape.matrix()
The basic function of enrichment analysis is done using the ```escape.matrix()``` function, with the user defining the **method** and **gene.sets** to use.
```r
#Defining Gene Set To Use:
GS <- list(Bcells = c("MS4A1", "CD79B", "CD79A", "IGH1", "IGH2"),
Tcells = c("CD3E", "CD3D", "CD3G", "CD7","CD8A"))
#Using Seurat Built-In Example:
pbmc_small <- SeuratObject::pbmc_small
#Running Enrichment
trial.ssGSEA <- escape.matrix(pbmc_small,
method = "ssGSEA",
gene.sets = GS,
min.size = NULL)
```
#### runEscape()
Alternatively, ```runEscape()``` will perform the enrichment calculations as above, but also automatically amend the single-cell object with the values added as an assay, which is named via the **new.assay.name** parameter. This facilitates easy downstream visualization and analysis.
```r
pbmc_small <- runEscape(pbmc_small,
method = "ssGSEA",
new.assay.name = "escape.ssGSEA",
gene.sets = GS,
min.size = NULL)
```
### Vignette
A more comprehensive vignette including the visualizations, principal component analysis and differential testing is available [here](https://www.borch.dev/uploads/screpertoire/articles/Running_Escape.html).
### Citation
If using escape, please cite the [article](https://www.nature.com/articles/s42003-020-01625-6): Borcherding, N., Vishwakarma, A., Voigt, A.P. et al. Mapping the immune environment in clear cell renal carcinoma by single-cell genomics. Commun Biol 4, 122 (2021).
### Contact
Questions, comments, suggestions, please feel free to contact Nick Borcherding via this repository, [email](mailto:ncborch@gmail.com), or using [twitter](https://twitter.com/theHumanBorch).
