Different sources: Legacy vs Harmonized
There are two available sources to download GDC data using TCGAbiolinks:
- GDC Legacy Archive : provides access to an unmodified copy of data that was previously stored in
[CGHub](https://cghub.ucsc.edu/) and in the TCGA Data Portal hosted by the TCGA Data Coordinating Center (DCC), in which uses
as references GRCh37 (hg19) and GRCh36 (hg18).
- GDC harmonized database: data available was harmonized against GRCh38 (hg38) using GDC Bioinformatics Pipelines
which provides methods to the standardization of biospecimen and
clinical data.
Understanding the barcode
A TCGA barcode is composed of a collection of identifiers. Each specifically identifies a TCGA data element. Refer to the following figure for an illustration of how metadata identifiers comprise a barcode. An aliquot barcode contains the highest number of identifiers.
Example:
- Aliquot barcode: TCGA-G4-6317-02A-11D-2064-05
- Participant: TCGA-G4-6317
- Sample: TCGA-G4-6317-02
For more information check [GDC TCGA barcodes](https://docs.gdc.cancer.gov/Encyclopedia/pages/TCGA_Barcode/)
# Searching arguments
You can easily search GDC data using the `GDCquery` function.
Using a summary of filters as used in the TCGA portal, the function works
with the following arguments:
| ?project | A list of valid project (see table below)] | |
|----------------------- |------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- |------------------------------------- |
| data.category | A valid project (see list with TCGAbiolinks:::getProjectSummary(project)) | |
| data.type | A data type to filter the files to download | |
| workflow.type | GDC workflow type | |
| legacy | Search in the legacy repository | |
| access | Filter by access type. Possible values: controlled, open | |
| platform | Example: | |
| | CGH- 1x1M_G4447A | IlluminaGA_RNASeqV2 |
| | AgilentG4502A_07 | IlluminaGA_mRNA_DGE |
| | Human1MDuo | HumanMethylation450 |
| | HG-CGH-415K_G4124A | IlluminaGA_miRNASeq |
| | HumanHap550 | IlluminaHiSeq_miRNASeq |
| | ABI | H-miRNA_8x15K |
| | HG-CGH-244A | SOLiD_DNASeq |
| | IlluminaDNAMethylation_OMA003_CPI | IlluminaGA_DNASeq_automated |
| | IlluminaDNAMethylation_OMA002_CPI | HG-U133_Plus_2 |
| | HuEx- 1_0-st-v2 | Mixed_DNASeq |
| | H-miRNA_8x15Kv2 | IlluminaGA_DNASeq_curated |
| | MDA_RPPA_Core | IlluminaHiSeq_TotalRNASeqV2 |
| | HT_HG-U133A | IlluminaHiSeq_DNASeq_automated |
| | diagnostic_images | microsat_i |
| | IlluminaHiSeq_RNASeq | SOLiD_DNASeq_curated |
| | IlluminaHiSeq_DNASeqC | Mixed_DNASeq_curated |
| | IlluminaGA_RNASeq | IlluminaGA_DNASeq_Cont_automated |
| | IlluminaGA_DNASeq | IlluminaHiSeq_WGBS |
| | pathology_reports | IlluminaHiSeq_DNASeq_Cont_automated |
| | Genome_Wide_SNP_6 | bio |
| | tissue_images | Mixed_DNASeq_automated |
| | HumanMethylation27 | Mixed_DNASeq_Cont_curated |
| | IlluminaHiSeq_RNASeqV2 | Mixed_DNASeq_Cont |
| file.type | To be used in the legacy database for some platforms, to define which file types to be used. | |
| barcode | A list of barcodes to filter the files to download | |
| experimental.strategy | Filter to experimental strategy. Harmonized: WXS, RNA-Seq, miRNA-Seq, Genotyping Array. Legacy: WXS, RNA-Seq, miRNA-Seq, Genotyping Array, DNA-Seq, Methylation array, Protein expression array, WXS,CGH array, VALIDATION, Gene expression array,WGS, MSI-Mono-Dinucleotide Assay, miRNA expression array, Mixed strategies, AMPLICON, Exon array, Total RNA-Seq, Capillary sequencing, Bisulfite-Seq | |
| sample.type | A sample type to filter the files to download | |
## project options
The options for the field `project` are below:
```{r, eval = TRUE, echo = FALSE}
datatable(
TCGAbiolinks:::getGDCprojects(),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 10),
rownames = FALSE,
caption = "List of projects"
)
```
## sample.type options
The options for the field `sample.type` are below:
```{r, eval = TRUE, echo = FALSE}
datatable(
TCGAbiolinks:::getBarcodeDefinition(),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 10),
rownames = FALSE,
caption = "List sample types"
)
```
The other fields (data.category, data.type, workflow.type, platform, file.type) can be found below.
Please, note that these tables are still incomplete.
## Harmonized data options (`legacy = FALSE`)
```{r}
datatable(readr::read_csv("https://docs.google.com/spreadsheets/d/1f98kFdj9mxVDc1dv4xTZdx8iWgUiDYO-qiFJINvmTZs/export?format=csv&gid=2046985454",col_types = readr::cols()),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 40),
rownames = FALSE)
```
## Legacy archive data options (`legacy = TRUE`)
```{r}
datatable(readr::read_csv("https://docs.google.com/spreadsheets/d/1f98kFdj9mxVDc1dv4xTZdx8iWgUiDYO-qiFJINvmTZs/export?format=csv&gid=1817673686",col_types = readr::cols()),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 40),
rownames = FALSE)
```
# Harmonized database examples
## DNA methylation data: Recurrent tumor samples
In this example we will access the harmonized database (`legacy = FALSE`)
and search for all DNA methylation data for recurrent glioblastoma multiform (GBM)
and low grade gliomas (LGG) samples.
```{r message=FALSE, warning=FALSE}
query <- GDCquery(
project = c("TCGA-GBM", "TCGA-LGG"),
data.category = "DNA Methylation",
legacy = FALSE,
platform = c("Illumina Human Methylation 450"),
sample.type = "Recurrent Tumor"
)
datatable(getResults(query),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 5),
rownames = FALSE)
```
## Samples with DNA methylation and gene expression data
In this example we will access the harmonized database (`legacy = FALSE`)
and search for all patients with DNA methylation (platform HumanMethylation450k) and gene expression data
for Colon Adenocarcinoma tumor (TCGA-COAD).
```{r message=FALSE, warning = FALSE, eval = FALSE}
query.met <- GDCquery(
project = "TCGA-COAD",
data.category = "DNA Methylation",
legacy = FALSE,
platform = c("Illumina Human Methylation 450")
)
query.exp <- GDCquery(
project = "TCGA-COAD",
data.category = "Transcriptome Profiling",
data.type = "Gene Expression Quantification",
workflow.type = "HTSeq - FPKM-UQ"
)
# Get all patients that have DNA methylation and gene expression.
common.patients <- intersect(
substr(getResults(query.met, cols = "cases"), 1, 12),
substr(getResults(query.exp, cols = "cases"), 1, 12)
)
# Only seelct the first 5 patients
query.met <- GDCquery(
project = "TCGA-COAD",
data.category = "DNA Methylation",
legacy = FALSE,
platform = c("Illumina Human Methylation 450"),
barcode = common.patients[1:5]
)
query.exp <- GDCquery(
project = "TCGA-COAD",
data.category = "Transcriptome Profiling",
data.type = "Gene Expression Quantification",
workflow.type = "HTSeq - FPKM-UQ",
barcode = common.patients[1:5]
)
```
```{r results_matched, message=FALSE, warning=FALSE, eval = FALSE}
datatable(
getResults(query.met, cols = c("data_type","cases")),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 5),
rownames = FALSE
)
datatable(
getResults(query.exp, cols = c("data_type","cases")),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 5),
rownames = FALSE
)
```
## Raw Sequencing Data: Finding the match between file names and barcode for Controlled data.
This exmaple shows how the user can search for breast cancer Raw Sequencing Data ("Controlled")
and verify the name of the files and the barcodes associated with it.
```{r message=FALSE, warning=FALSE}
query <- GDCquery(
project = c("TCGA-BRCA"),
data.category = "Sequencing Reads",
sample.type = "Primary Tumor"
)
# Only first 100 to make render faster
datatable(
getResults(query, rows = 1:100,cols = c("file_name","cases")),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 5),
rownames = FALSE
)
```
# Legacy archive examples
## DNA methylation
### Array-based assays
This example shows how the user can search for glioblastoma multiform (GBM)
and DNA methylation data
for platform Illumina Human Methylation 450 and Illumina Human Methylation 27.
```{r message=FALSE, warning=FALSE}
query <- GDCquery(
project = c("TCGA-GBM"),
legacy = TRUE,
data.category = "DNA methylation",
platform = c("Illumina Human Methylation 450", "Illumina Human Methylation 27")
)
datatable(
getResults(query, rows = 1:100),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 5),
rownames = FALSE
)
```
### whole-genome bisulfite sequencing (WGBS)
```{r message = FALSE, warning = FALSE, eval = FALSE}
query <- GDCquery(
project = c("TCGA-LUAD"),
legacy = TRUE,
data.category = "DNA methylation",
data.type = "Methylation percentage",
experimental.strategy = "Bisulfite-Seq"
)
# VCF - controlled data
query <- GDCquery(
project = c("TCGA-LUAD"),
legacy = TRUE,
data.category = "DNA methylation",
data.type = "Bisulfite sequence alignment",
experimental.strategy = "Bisulfite-Seq"
)
# WGBS BAM files - controlled data
query <- GDCquery(
project = c("TCGA-LUAD"),
legacy = TRUE,
data.type = "Aligned reads",
data.category = "Raw sequencing data",
experimental.strategy = "Bisulfite-Seq"
)
```
## Gene expression
This exmaple shows how the user can search for glioblastoma multiform (GBM)
gene expression data with the normalized results for expression of a gene.
For more information about file.types check [GDC TCGA file types](https://gdc.cancer.gov/resources-tcga-users/legacy-archive-tcga-tag-descriptions)
```{r message=FALSE, warning=FALSE}
# Gene expression aligned against hg19.
query.exp.hg19 <- GDCquery(
project = "TCGA-GBM",
data.category = "Gene expression",
data.type = "Gene expression quantification",
platform = "Illumina HiSeq",
file.type = "normalized_results",
experimental.strategy = "RNA-Seq",
barcode = c("TCGA-14-0736-02A-01R-2005-01", "TCGA-06-0211-02A-02R-2005-01"),
legacy = TRUE
)
datatable(
getResults(query.exp.hg19),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 5),
rownames = FALSE
)
```
# Get Manifest file
If you want to get the manifest file from the query object you can use the function *getManifest*. If you
set save to TRUEm a txt file that can be used with GDC-client Data transfer tool (DTT) or with its GUI version [ddt-ui](https://github.com/NCI-GDC/dtt-ui) will be created.
```{r message=FALSE, warning=FALSE}
getManifest(query.exp.hg19,save = FALSE)
```
# ATAC-seq data
For the moment, ATAC-seq data is available at the [GDC publication page](https://gdc.cancer.gov/about-data/publications/ATACseq-AWG).
Also, for more details, you can check an ATAC-seq workshop at http://rpubs.com/tiagochst/atac_seq_workshop
The list of file available is below:
```{r message=FALSE, warning=FALSE}
datatable(
getResults(TCGAbiolinks:::GDCquery_ATAC_seq())[,c("file_name","file_size")],
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 5),
rownames = FALSE
)
```
You can use the function `GDCquery_ATAC_seq` filter the manifest table and use `GDCdownload` to save the data locally.
```{r message=FALSE, warning=FALSE,eval = FALSE}
query <- TCGAbiolinks:::GDCquery_ATAC_seq(file.type = "rds")
GDCdownload(query,method = "client")
query <- TCGAbiolinks:::GDCquery_ATAC_seq(file.type = "bigWigs")
GDCdownload(query,method = "client")
```
# Summary of available files per patient
Retrieve the numner of files under each data_category + data_type + experimental_strategy + platform.
Almost like https://portal.gdc.cancer.gov/exploration
```{r message=FALSE, warning=FALSE,eval = TRUE}
tab <- getSampleFilesSummary(project = "TCGA-ACC")
datatable(
head(tab),
filter = 'top',
options = list(scrollX = TRUE, keys = TRUE, pageLength = 5),
rownames = FALSE
)
```