# drugfindR

    library(tidyverse)
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    #> ℹ Use the conflicted package (<http://conflicted.r-lib.org/>) to force all conflicts to become errors
    library(drugfindR)

## Introduction

`drugfindR` provides end-users with a convenient method for accessing the Library of Integrated Network-Based Cellular Signatures (LINCS). The LINCS project aims to create a network-based understanding of biology by systematically cataloging changes in cellular processes, namely gene expression, that occur when cells are exposed to a variety of perturbing agents. iLINCS is an integrated web-based platform designed for the analysis of omics data and signatures of cellular perturbagens. While the iLINCS analysis workflows integrate vast omics data resources and a range of analytic and visual tools into a comprehensive platform, `drugfindR` is advantageous in that it is scriptable and usable from within R without relying on the iLINCS web platform. `drugfindR` also possesses the capability of running all input signatures simultaneously, which makes investigating a particular gene or drug extremely efficient. From the output data generated by `drugfindR`, end-users may understand how the overexpression or knockdown of a specific gene affects the expression of genes within the same cellular system, identify downstream molecular consequences of gene perturbation within a system, and investigate candidate drugs that may be repurposed for other physiological reasons.

## Installation

`drugfindR` can be installed from BioConductor using the `BiocManager` package:

    BiocManager::install("drugfindR") # nolint: nonportable_path_linter.

## Use Cases

`drugfindR` has multiple features that make interfacing with the iLINCS database and analyzing LINCS data simple and efficient. However, the package is explicitly designed for two primary use cases:

1. Using an input transcriptomic signature to identify candidate drugs in the iLINCS database
2. Identifying drugs or other genes that are similar (or opposite) in function to a given drug or gene.

## Package Design

This package provides two different ways to achieve these use cases. First, there is a set of five functions that can be deployed in a pipeline for the results. Then, there are two functions `investigateTarget()` and `investigateSignature()` that perform the entire pipeline in one function call with sensible defaults.

### Pipeline Components

The five pipeline functions are:

1. `getSignature()`: This function takes a LINCS ID and returns the corresponding signature.
2. `prepareSignature()`: This function takes a transcriptomic signature and prepares it for analysis by `drugfindR`.
3. `filterSignature()`: This function takes a signature and filters it to given thresholds.
4. `getConcordants()`: This function takes a signature and returns the concordant signatures from the iLINCS database.
5. `consensusConcordants()`: This function takes a list of concordant signatures and returns a list of consensus signature.

## Use Case 1: Identifying Candidate Drugs from an Input Signature

For this case, we will use one of the signatures that was used in the paper “Identification of candidate repurposable drugs to combat COVID - 19 using a signature - based approach” by O’Donovan, Imami, et al.

In that paper, the authors used the available gene expression data from cells infected with SARS-CoV-2 to identify potential drugs that could be repurposed to treat COVID-19. We will use one of the signatures that they have provided in their paper to showcase how `drugfindR` can be used to identify candidate drugs from an input signature. We will use the `dCovid_diffexp.tsv` signature from the paper.

### Step 1: Get the Signature

This signature is available with the package. Our first step is to import the signature so we can work with it. The `read_tsv()` function from the `readr` package can be used to read the signature into R from a remote URL or a local file.

    # Load the signature from the paper

    diffexp <- read_tsv(
        system.file("extdata", "dCovid_diffexp.tsv",
            package = "drugfindR"
        )
    )
    #> Rows: 4090 Columns: 3
    #> ── Column specification ────────────────────────────────────────────────────────
    #> Delimiter: "\t"
    #> chr (1): hgnc_symbol
    #> dbl (2): logFC, PValue
    #>
    #> ℹ Use `spec()` to retrieve the full column specification for this data.
    #> ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.

    # Take a look at the signature

    head(diffexp) |>
        knitr::kable()

hgnc_symbol | logFC | PValue
--- | --- | ---
CCL4L2 | -3.976903 | 1.80e-06
IL5RA | -4.830082 | 8.70e-06
FN1 | 4.942690 | 1.17e-05
GSTM1 | -8.211472 | 1.53e-05
CD180 | 2.153878 | 2.02e-05
FAM20C | 3.112871 | 2.55e-05

We can see that the signature has `ncol(diffexp)` columns and `nrow(diffexp)` rows. The names of the columns are typical of what you would get from edgeR or DESeq2.

### Step 2: Prepare the Signature

The next step is to prepare the signature for analysis by `drugfindR`. This step is necessary because the signature can be in many different formats, with different names for columns. iLINCS needs columns to be in a specific order and with specific names. The `prepareSignature()` function takes care of this for us.

`prepareSignature()` takes three optional arguments:

1. `geneColumn`: The name of the column in the input that contains the gene names. The default is `"Symbol"`.
2. `logfcColumn`: The name of the column in the input that contains the log fold change values. The default is `"logFC"`.
3. `pvalColumn`: The name of the column in the input that contains the p-values. The default is `"PValue"`.

    # Prepare the signature for analysis
    # The only thing that is different from the defaults is the gene_column
    # so we will specify that and rely on defaults for others.

    signature <- prepareSignature(diffexp,
        geneColumn = "hgnc_symbol"
    )

    # Take a look at the signature

    head(signature) |>
        knitr::kable()

| signatureID | ID_geneid | Name_GeneSymbol | Value_LogDiffExp | Significance_pvalue
--- | --- | --- | --- | --- | ---
1 | InputSig | 4860 | PNP | 1.709692 | 0.0024364
5 | InputSig | 4125 | MAN2B1 | 1.100506 | 0.0030275
8 | InputSig | 2274 | FHL2 | 1.330287 | 0.1423757
14 | InputSig | 351 | APP | 1.050513 | 0.0108441
26 | InputSig | 7077 | TIMP2 | 1.795990 | 0.0124167
29 | InputSig | 23659 | PLA2G15 | 1.113303 | 0.0495367

We can see that the signature has been reordered and renamed. The first column is now `names(signature)[1]`, the second column is now `names(signature)[2]`, and the third column is now `names(signature)[3]`, which is what iLINCS expects.

### Step 3: Filter the Signature

Now that we have the signature in the correct format and filtered to the L1000 genes, we can filter it to the thresholds that we want. This filter step is necessary because we would like to use the genes that have a high enough change for it to matter.

The `filterSignature()` function can filter based on logFC values in two ways:

1. Absolute Threshold: You can give an absolute threshold ( or a pair of absolute thresholds) for the logFC values. Any genes that do not meet the threshold will be removed from the signature.

2. Percentile Threshold: You can give a percentile threshold (or a pair of percentile thresholds) for the logFC values. Any genes that do not meet the threshold will be removed from the signature.

The `filterSignature()` function takes three arguments:

1. `signature`: The signature to filter.
2. `direction`: This argument specifies whether to filter for upregulated genes, downregulated genes, or both. The default is `"any"`.
3. One of `threshold` or `prop`: The threshold argument is used to specify an absolute threshold (or a pair of absolute thresholds) for the logFC values. The prop argument is used to specify a percentile threshold (or a pair of percentile thresholds) for the logFC values. They can not be specified together.

    # Filter the signature to only include genes that are upregulated by at least
    # 1.5 logFC

    filteredSignatureUp <- filterSignature(signature,
        direction = "up",
        threshold = 1.5
    )

    filteredSignatureUp |>
        head() |>
        knitr::kable()

signatureID | ID_geneid | Name_GeneSymbol | Value_LogDiffExp | Significance_pvalue
--- | --- | --- | --- | ---
InputSig | 4860 | PNP | 1.709692 | 0.0024364
InputSig | 7077 | TIMP2 | 1.795990 | 0.0124167
InputSig | 890 | CCNA2 | 1.750080 | 0.0163805
InputSig | 991 | CDC20 | 3.399809 | 0.0003236
InputSig | 9093 | DNAJA3 | 2.340929 | 0.0199756
InputSig | 1509 | CTSD | 1.547209 | 0.0019360

    # Filter the signature to only include genes that are downregulated by at least
    # 1.5 logFC
    filteredSignatureDn <- filterSignature(signature,
        direction = "down",
        threshold = 1.5
    )

    filteredSignatureDn |>
        head() |>
        knitr::kable()

signatureID | ID_geneid | Name_GeneSymbol | Value_LogDiffExp | Significance_pvalue
--- | --- | --- | --- | ---
InputSig | 2810 | SFN | -2.056475 | 0.0151082
InputSig | 3815 | KIT | -1.635917 | 0.0225406
InputSig | 2625 | GATA3 | -2.310024 | 0.0018368
InputSig | 10123 | ARL4C | -1.638483 | 0.0005734
InputSig | 2624 | GATA2 | -2.728454 | 0.0110850
InputSig | 2946 | GSTM2 | -1.666703 | 0.0055149

### Step 4: Get the Concordant Signatures

Now that we have the filtered signatures for both upregulated and downregulated genes, we can get the concordant signatures from the iLINCS database. The `getConcordants()` function takes a signature and returns the concordant signatures from the iLINCS database. It also requires specification of the database to target for the concordant signatures.

The `getConcordants()` function takes the following arguments:

1. `signature`: The signature to get concordant signatures for.
2. `ilincsLibrary`: The iLINCS library to target for concordant signatures. This can be one of c(“OE”, “KD”, “CP”), standing for overexpression, knockdown, and chemical perturbagens, respectively.
3. `direction`: This argument specifies whether the input signature is upregulated or downregulated. This is useful to annotate the output. This is `NULL` by default.

    # Get the concordant signatures for the upregulated signature

    upConcordants <- getConcordants(filteredSignatureUp, ilincsLibrary = "CP")

    upConcordants |>
        head() |>
        knitr::kable()

signatureid | treatment | concentration | time | cellline | similarity | pValue | sig_direction | sig_type
--- | --- | --- | --- | --- | --- | --- | --- | ---
LINCSCP_99767 | Mitoxantrone | 0.04uM | 24h | HCC515 | 0.6259367 | 0e+00 | Up | Chemical Perturbagen
LINCSCP_207881 | Navitoclax | 10uM | 6h | H1299 | 0.6121090 | 0e+00 | Up | Chemical Perturbagen
LINCSCP_207785 | Myriocin | 10uM | 6h | H1299 | 0.5997346 | 0e+00 | Up | Chemical Perturbagen
LINCSCP_285868 | CHEMBL259628 | 10uM | 6h | SKM1 | 0.5835506 | 1e-07 | Up | Chemical Perturbagen
LINCSCP_163143 | Crizotinib | 0.12uM | 3h | SKBR3 | 0.5759650 | 1e-07 | Up | Chemical Perturbagen
LINCSCP_64976 | PP-242 | 0.5uM | 24h | VCAP | -0.5674210 | 2e-07 | Up | Chemical Perturbagen

    # Get the concordant signatures for the downregulated signature

    dnConcordants <- getConcordants(filteredSignatureDn, ilincsLibrary = "CP")

    dnConcordants |>
        head() |>
        knitr::kable()

signatureid | treatment | concentration | time | cellline | similarity | pValue | sig_direction | sig_type
--- | --- | --- | --- | --- | --- | --- | --- | ---
LINCSCP_260147 | PD-98059 | 10uM | 6h | NEU | 0.9276283 | 6.00e-07 | Down | Chemical Perturbagen
LINCSCP_149954 | 848193-68-0 | 0.12uM | 24h | NPC.TAK | -0.9235333 | 9.00e-07 | Down | Chemical Perturbagen
LINCSCP_40982 | Devazepide | 10uM | 24h | NEU | 0.9133641 | 2.00e-06 | Down | Chemical Perturbagen
LINCSCP_292245 | MLS003568137 | 10uM | 24h | VCAP | -0.8890042 | 9.30e-06 | Down | Chemical Perturbagen
LINCSCP_150380 | VX-745 | 0.04uM | 24h | NPC.TAK | 0.8854791 | 1.13e-05 | Down | Chemical Perturbagen
LINCSCP_150801 | Mitoxantrone | 1.11uM | 24h | PC3 | 0.8736419 | 2.09e-05 | Down | Chemical Perturbagen

### Step 5: Get the list of Consensus Concordant Signatures

Now that we have the concordant signatures for both the upregulated and downregulated signatures, we can get the list of consensus concordant signatures. The `consensusConcordants()` function takes a list of concordant signatures and returns a list of consensus signatures. This function also takes a number of optional arguments that can be used to control the consensus list generation.

By default the consensus list performs the following steps:

1. Combine the list of concordant signatures into a single data frame.
2. For each individual signature origin (Gene or Drug), choose the one with the largest absolute concordance value.

Additionally, we can filter by the cell line to only include the cell lines of interest.

The `consensusConcordants()` function takes the following arguments:

1. `...`: One or Two (see paired) Data Frames with the concordants
2. `paired`: A logical value indicating whether the input is a single data frame with paired signatures or two data frames with unpaired signatures. The default is `FALSE`.
3. `cellLines`: A character vector of cell lines to filter the consensus list to. The default is `NULL`, which means no filtering.
4. `cutoff`: The absolute cutoff value of similarity to use when filtering the consensus list. The default is `0.321`.

    # Get the consensus concordant signatures for the upregulated signature

    consensus <- consensusConcordants(upConcordants, dnConcordants,
        paired = TRUE, cutoff = 0.2
    )

    consensus |>
        head() |>
        knitr::kable()

TargetSignature | Target | TargetCellLine | TargetTime | TargetConcentration | InputSigDirection | SignatureType | Similarity | pValue
--- | --- | --- | --- | --- | --- | --- | --- | ---
LINCSCP_260147 | PD-98059 | NEU | 6h | 10uM | Down | Chemical Perturbagen | 0.9276283 | 6.00e-07
LINCSCP_149954 | 848193-68-0 | NPC.TAK | 24h | 0.12uM | Down | Chemical Perturbagen | -0.9235333 | 9.00e-07
LINCSCP_40982 | Devazepide | NEU | 24h | 10uM | Down | Chemical Perturbagen | 0.9133641 | 2.00e-06
LINCSCP_292245 | MLS003568137 | VCAP | 24h | 10uM | Down | Chemical Perturbagen | -0.8890042 | 9.30e-06
LINCSCP_150380 | VX-745 | NPC.TAK | 24h | 0.04uM | Down | Chemical Perturbagen | 0.8854791 | 1.13e-05
LINCSCP_150801 | Mitoxantrone | PC3 | 24h | 1.11uM | Down | Chemical Perturbagen | 0.8736419 | 2.09e-05

### Alternate One-Step Method

The above method breaks down the entire method into five steps. However, `drugfindR` also provides two functions that perform the entire pipeline in one function call with sensible defaults. These functions are `investigateTarget()` and `investigateSignature()`.

For this use case, `investigateSignature()` is the function that we want to use. It takes the following required arguments:

1. `expr`: The signature to investigate.
2. `outputLib`: The iLINCS library to target for concordant signatures. This can be one of c(“OE”, “KD”, “CP”), standing for overexpression, knockdown, and chemical perturbagens, respectively.
3. `filterThreshold`: The absolute threshold (or a pair of absolute thresholds) for the logFC values. Any genes that do not meet the threshold will be removed from the signature.
4. `filterProp`: The percentile threshold (or a pair of percentile thresholds) for the logFC values. Any genes that do not meet the threshold will be removed from the signature.

Other arguments that have sensible defaults are:

1. `similarityThreshold`: The absolute cutoff value of similarity to use when filtering the consensus list. The default is `0.2`.
2. `paired`: A logical value indicating whether the to split the input dataframe in up and downregulated signatures. The default is `TRUE`.
3. `outputCellLines`: A character vector of cell lines to filter the consensus list to. The default is `NULL`, which means no filtering.
4. `geneColumn`: The name of the column in the input that contains the gene names. The default is `"Symbol"`.
5. `logfcColumn`: The name of the column in the input that contains the log fold change values. The default is `"logFC"`.
6. `pvalColumn`: The name of the column in the input that contains the p-values. The default is `"PValue"`.
7. `sourceName`: The name of the source of the signature. The default is `"Input"`.
8. `sourceCellLine`: The cell line of the source of the signature. The default is `NA`.
9. `sourceTime`: The time of the source of the signature. The default is `NA`.
10. `sourceConcentration`: The concentration of the source of the signature. The default is `NA`.

    investigated <- investigateSignature(diffexp,
        outputLib = "CP", filterThreshold = 1.5,
        geneColumn = "hgnc_symbol", logfcColumn = "logFC",
        pvalColumn = "PValue"
    )

    investigated |>
        head() |>
        knitr::kable()

Source | Target | Similarity | SourceSignature | SourceCellLine | SourceConcentration | SourceTime | TargetSignature | TargetCellLine | TargetConcentration | TargetTime | InputSigDirection | SignatureType | pValue
--- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | ---
Input | PD-98059 | 0.9276283 | InputSig | NA | NA | NA | LINCSCP_260147 | NEU | 10uM | 6h | Down | Chemical Perturbagen | 6.00e-07
Input | 848193-68-0 | -0.9235333 | InputSig | NA | NA | NA | LINCSCP_149954 | NPC.TAK | 0.12uM | 24h | Down | Chemical Perturbagen | 9.00e-07
Input | Devazepide | 0.9133641 | InputSig | NA | NA | NA | LINCSCP_40982 | NEU | 10uM | 24h | Down | Chemical Perturbagen | 2.00e-06
Input | MLS003568137 | -0.8890042 | InputSig | NA | NA | NA | LINCSCP_292245 | VCAP | 10uM | 24h | Down | Chemical Perturbagen | 9.30e-06
Input | VX-745 | 0.8854791 | InputSig | NA | NA | NA | LINCSCP_150380 | NPC.TAK | 0.04uM | 24h | Down | Chemical Perturbagen | 1.13e-05
Input | Mitoxantrone | 0.8736419 | InputSig | NA | NA | NA | LINCSCP_150801 | PC3 | 1.11uM | 24h | Down | Chemical Perturbagen | 2.09e-05

## Environment Setup

    devtools::session_info()
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    #>  setting  value
    #>  version  R version 4.5.2 (2025-10-31)
    #>  os       Ubuntu 24.04.3 LTS
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    #>  tz       Etc/UTC
    #>  date     2026-01-15
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    #>  quarto   1.8.24 @ /usr/local/bin/quarto
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    #>  usethis        3.2.1     2025-09-06 [2] RSPM (R 4.5.0)
    #>  vctrs          0.6.5     2023-12-01 [2] RSPM (R 4.5.0)
    #>  vroom          1.6.7     2025-11-28 [2] RSPM (R 4.5.0)
    #>  withr          3.0.2     2024-10-28 [2] RSPM (R 4.5.0)
    #>  xfun           0.55      2025-12-16 [2] RSPM (R 4.5.0)
    #>  yaml           2.3.12    2025-12-10 [2] RSPM (R 4.5.0)
    #>
    #>  [1] /tmp/RtmppGvIiL/Rinst1ad74d73a3dc
    #>  [2] /github/workspace/pkglib
    #>  [3] /usr/local/lib/R/site-library
    #>  [4] /usr/lib/R/site-library
    #>  [5] /usr/lib/R/library
    #>  * ── Packages attached to the search path.
    #>
    #> ──────────────────────────────────────────────────────────────────────────────