From f54b0d327ae741507c9f63f0824df1a6fb9a4bbb Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 11:58:27 -0400 Subject: [PATCH 01/18] update Readme --- README.md | 54 ++++++++++++++++++++++++++++++++++++++++-------------- 1 file changed, 40 insertions(+), 14 deletions(-) diff --git a/README.md b/README.md index 665341c..6bbd89c 100644 --- a/README.md +++ b/README.md @@ -1,31 +1,47 @@ -# D3b: Loss of Heterozygosity (LOH) +# Kids First Loss of Heterozygosity (LOH) -This CWL workflow assesses the loss of heterozygosity(LOH) in the tumor for rare germline variants (gnomad_3_1_1_AF_popmax < 0.01 or gnomad_3_1_1_AF_popmax == '.'). This workflow has been deployed on [cavatica](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/apps/Loss_of_Heterozygosity). It was tested with a [proband only run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/522d128a-2195-4c9c-8339-1709da16821d/) and a [trio run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/d7f6b667-35ef-46a7-a666-970a78ef3175/) +![data service logo](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/kfdrc-logo-sm.png) -### Repo Description +The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This workflow is designed to analyze LOH for family trios as well as multiple proband tumor samples. -This workflow is divided into two tools: Germline tool and tumor tool. + This workflow has been deployed on [cavatica](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/apps/Loss_of_Heterozygosity). It was tested with a [proband only run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/522d128a-2195-4c9c-8339-1709da16821d/) and a [trio run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/d7f6b667-35ef-46a7-a666-970a78ef3175/) -Here are the basic steps for the LOH assessment workflow: +### App Description -* Filter germline annotations to retain variants with gnomad_3_1_1_AF_popmax < 0.01 or gnomad_3_1_1_AF_popmax not defined performed by the germline tool (this gets us rare germline variants). -* Gather variant information (gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, VAF) covered by the germline tool. -* Search in a paired tumor sample (example match to above germline) and, if applicable, parental germline samples, for the same variant and calculate the VAF covered by tumor tool. -* LOH will create an output file with headers (BS_id, gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, VAF) for each of: proband germline, proband tumor, paternal germline, maternal germline covered by the tumor tool. +The Kids First Loss of Heterozygosity app is divided into two tools: Germline tool and tumor tool. +#### Germline tool -(Optional) This workflow has the ability to analyze LOH for trios, with multiple tumor samples. The input requirement for family trios is a trio vcf, a peddy file, and a tumor cram file for the proband. Note: The user can provide multiple .cram or.bam files. +Germline tool filters germline annotations to retain variants based on gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. It requires vcf file, proband sample id, ram as required inputs and peddy file as optional input which is required for family trios. It outputs variant information such as gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, variant allele frequency and list of coordinates that will be an input to tumor tool. -### Running it locally on a laptop? +#### Tumor tool +Tumor tool search in paired proband tumor sample for aligned reads in the regions where rare variants from the germline tool exists and exact allele/reference count, allele/reference depth and calculate the variant allele frequency VAF. Tumor tool have the capability to search multiple tumor samples for proband and if applicable, parental and maternal tumor samples. To exact reads from the bam/cram files, this tool utilizes [bam-readcount](https://github.com/genome/bam-readcount) and wraps it with python script to shape the output in a tabular format. +#### Inputs +``` +Germline tool + BS_ID: { doc: provide BS id for germline normal,type: string } + frequency: { doc: provide popmax cutoff for rare germline variants, type: 'float?', default: 0.01 } + ram_germline: { doc: Provide ram (in GB) based on the size of vcf,type: 'int?', default: 8} + peddy_file: { doc: provide ped file for the trio, type: 'File?' } +Tumor tool + participant_id: { doc: provide participant id for this run, type: string } + bamscrams: { doc: tumor input file in cram or bam format with their index file, type: 'File[]' , secondaryFiles: [ { pattern: ".crai", required: false }, { pattern: ".bai", required: false } ] } + minDepth: { doc: provide minDepth to consider for tumor reads, type: 'int?', default: 1 } + reference: { doc: human reference in fasta format with index file, type: File,secondaryFiles: [ .fai ] } + sample_vcf_file: { doc: provide germline vcf file for this sample, type: File } + bamcramsampleIDs: { doc: provide unique identifers (in the same order) for cram/bam files provided under bamcrams tag. Default is sample ID pulled from bam/cram files., type: 'string[]?' } + ram_tumor: { doc: Provide ram (in GB) size and number of cram/bam inputs, type: 'int?', default: 16} + minCore: { type: 'int?', default: 16, doc: "Minimum number of cores for tumor tool" } +``` -It is recommended to run this workflow on a system with a high number of CPUs and memory (>=16 GB). The basic requirement is a running docker engine and CWL tools. Command line to run the LOH workflow locally is: +#### Output ``` -cwltool workflow/run_LOH_app.cwl sample_input.yml +output_file: { type: File, doc: output file from LOH app, outputSource: run_tumor_tool/loh_output_file_tool } + ``` -Note: Inputs to the workflow need to be defined in sample_input.yml. ### Output headers @@ -55,3 +71,13 @@ LOH workflow will generate a output file with following headers: | proband_sample_id_tumor_depth | Depth of coverage for specific tumor sample | | proband_sample_id_tumor_alt_depth | Allele count at site for proband for specific tumor sample| | proband_sample_id_tumor_ref_depth | Reference count at site for proband for specific tumor sample | + +### Running it locally on a laptop? + + +It is recommended to run this workflow on a system with a high number of CPUs and memory (>=16 GB). The basic requirement is a running docker engine and CWL tools. Command line to run the LOH workflow locally is: + +``` +cwltool workflow/run_LOH_app.cwl sample_input.yml +``` +Note: Inputs to the workflow need to be defined in sample_input.yml. \ No newline at end of file From 0c405e45a6db1d6654e0728bed944a93dc78a2d5 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 12:13:41 -0400 Subject: [PATCH 02/18] update Readme --- README.md | 50 +++++++++++++++++++++++++------------------------- 1 file changed, 25 insertions(+), 25 deletions(-) diff --git a/README.md b/README.md index 6bbd89c..3479665 100644 --- a/README.md +++ b/README.md @@ -4,21 +4,23 @@ The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This workflow is designed to analyze LOH for family trios as well as multiple proband tumor samples. - This workflow has been deployed on [cavatica](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/apps/Loss_of_Heterozygosity). It was tested with a [proband only run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/522d128a-2195-4c9c-8339-1709da16821d/) and a [trio run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/d7f6b667-35ef-46a7-a666-970a78ef3175/) +Moreover, this workflow has been deployed on [cavatica](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/apps/Loss_of_Heterozygosity). It was tested with a [proband only run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/522d128a-2195-4c9c-8339-1709da16821d/) and a [trio run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/d7f6b667-35ef-46a7-a666-970a78ef3175/) -### App Description +### Application Description -The Kids First Loss of Heterozygosity app is divided into two tools: Germline tool and tumor tool. +The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and tumor tool. -#### Germline tool +#### Docker +Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH -Germline tool filters germline annotations to retain variants based on gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. It requires vcf file, proband sample id, ram as required inputs and peddy file as optional input which is required for family trios. It outputs variant information such as gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, variant allele frequency and list of coordinates that will be an input to tumor tool. +#### Germline Tool +Germline tool filters germline annotations to retain variants based on gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. It requires vcf file, proband sample id, ram as required inputs and peddy file as optional input which is required for family trios. It outputs variant information such as gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, variant allele frequency and list of coordinates that will be an input to tumor tool. -#### Tumor tool +#### Tumor Tool Tumor tool search in paired proband tumor sample for aligned reads in the regions where rare variants from the germline tool exists and exact allele/reference count, allele/reference depth and calculate the variant allele frequency VAF. Tumor tool have the capability to search multiple tumor samples for proband and if applicable, parental and maternal tumor samples. To exact reads from the bam/cram files, this tool utilizes [bam-readcount](https://github.com/genome/bam-readcount) and wraps it with python script to shape the output in a tabular format. -#### Inputs +### LOH Inputs ``` Germline tool BS_ID: { doc: provide BS id for germline normal,type: string } @@ -36,16 +38,15 @@ Tumor tool minCore: { type: 'int?', default: 16, doc: "Minimum number of cores for tumor tool" } ``` -#### Output +### LOH Output ``` output_file: { type: File, doc: output file from LOH app, outputSource: run_tumor_tool/loh_output_file_tool } - ``` ### Output headers -LOH workflow will generate a output file with following headers: +LOH workflow will generate a tab-separated values file with following headers and their description: | Headers | Description | |:-------:|:--------:| | BS_ID | Sample Id for germline sample | @@ -55,26 +56,25 @@ LOH workflow will generate a output file with following headers: | end | End position | | ref | Reference allele | | alt | Alternate Allele | -| proband_germline_ref_depth | Reference depth from germline for proband | -| proband_germline_alt_depth | Alternate depth from germline for proband | -| proband_germline_depth | Total number of reads overlapping a site for proband | -| proband_germline_vaf | Fraction of reads with the alternate allele for proband | -| paternal_germline_ref_depth | Reference depth from germline for father | -| paternal_germline_alt_depth | Alternate depth from germline for father | -| paternal_germline_depth | Total number of reads overlapping a site for father | -| paternal_germline_vaf | Fraction of reads with the alternate allele for father | -| maternal_germline_ref_depth | Reference depth from germline for mother | -| maternal_germline_alt_depth | Alternate depth from germline for mother | -| maternal_germline_depth | Total number of reads overlapping a site for mother | -| maternal_germline_vaf | Fraction of reads with the alternate allele for mother | -| proband_sample_id_tumor_vaf | Variant Allele frequency from tumor for proband for specific tumor sample| -| proband_sample_id_tumor_depth | Depth of coverage for specific tumor sample | +| proband_germline_ref_depth | Reference depth from germline from germline proband | +| proband_germline_alt_depth | Alternate depth from germline from germline proband | +| proband_germline_depth | Total number of reads overlapping a site from germline proband | +| proband_germline_vaf | Fraction of reads with the alternate allele from germline proband | +| paternal_germline_ref_depth | Reference depth from germline from paternal germline | +| paternal_germline_alt_depth | Alternate depth from germline from paternal germline | +| paternal_germline_depth | Total number of reads overlapping a site from paternal germline | +| paternal_germline_vaf | Fraction of reads with the alternate allele from paternal germline | +| maternal_germline_ref_depth | Reference depth from maternal germline | +| maternal_germline_alt_depth | Alternate depth from maternal germline | +| maternal_germline_depth | Total number of reads overlapping a site from mother | +| maternal_germline_vaf | Fraction of reads with the alternate allele from mother | +| proband_sample_id_tumor_vaf | Proband variant allele frequency from specific tumor sample| +| proband_sample_id_tumor_depth | Depth of coverage from specific tumor sample | | proband_sample_id_tumor_alt_depth | Allele count at site for proband for specific tumor sample| | proband_sample_id_tumor_ref_depth | Reference count at site for proband for specific tumor sample | ### Running it locally on a laptop? - It is recommended to run this workflow on a system with a high number of CPUs and memory (>=16 GB). The basic requirement is a running docker engine and CWL tools. Command line to run the LOH workflow locally is: ``` From 322a5402e1ac61ef8f087cc72c7630aa7c7862d2 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 12:43:55 -0400 Subject: [PATCH 03/18] Add image --- README.md | 22 +++++++++++++++------- docs/logo/loh.png | Bin 0 -> 105063 bytes 2 files changed, 15 insertions(+), 7 deletions(-) create mode 100644 docs/logo/loh.png diff --git a/README.md b/README.md index 3479665..26f74dd 100644 --- a/README.md +++ b/README.md @@ -23,30 +23,36 @@ Tumor tool search in paired proband tumor sample for aligned reads in the region ### LOH Inputs ``` Germline tool + # Required BS_ID: { doc: provide BS id for germline normal,type: string } frequency: { doc: provide popmax cutoff for rare germline variants, type: 'float?', default: 0.01 } + # Optional ram_germline: { doc: Provide ram (in GB) based on the size of vcf,type: 'int?', default: 8} + # Required for family trios otherwise not required peddy_file: { doc: provide ped file for the trio, type: 'File?' } Tumor tool + # Required participant_id: { doc: provide participant id for this run, type: string } bamscrams: { doc: tumor input file in cram or bam format with their index file, type: 'File[]' , secondaryFiles: [ { pattern: ".crai", required: false }, { pattern: ".bai", required: false } ] } - minDepth: { doc: provide minDepth to consider for tumor reads, type: 'int?', default: 1 } reference: { doc: human reference in fasta format with index file, type: File,secondaryFiles: [ .fai ] } sample_vcf_file: { doc: provide germline vcf file for this sample, type: File } + # Optional + minDepth: { doc: provide minDepth to consider for tumor reads, type: 'int?', default: 1 } bamcramsampleIDs: { doc: provide unique identifers (in the same order) for cram/bam files provided under bamcrams tag. Default is sample ID pulled from bam/cram files., type: 'string[]?' } - ram_tumor: { doc: Provide ram (in GB) size and number of cram/bam inputs, type: 'int?', default: 16} - minCore: { type: 'int?', default: 16, doc: "Minimum number of cores for tumor tool" } + ram_tumor: { doc: Provide ram (in GB) for tumor tool based on the number cram/bam inputs, type: 'int?', default: 16} + minCore: { type: 'int?', default: 16, doc: "Minimum number of cores for tumor tool based on the number cram/bam inputs" } ``` ### LOH Output +LOH application will output a tab-separated values file mapped data from germline tool and tumor tool. ``` output_file: { type: File, doc: output file from LOH app, outputSource: run_tumor_tool/loh_output_file_tool } ``` -### Output headers +#### Output headers -LOH workflow will generate a tab-separated values file with following headers and their description: +LOH workflow will generate a tab-separated values file with following headers: | Headers | Description | |:-------:|:--------:| | BS_ID | Sample Id for germline sample | @@ -70,8 +76,10 @@ LOH workflow will generate a tab-separated values file with following headers an | maternal_germline_vaf | Fraction of reads with the alternate allele from mother | | proband_sample_id_tumor_vaf | Proband variant allele frequency from specific tumor sample| | proband_sample_id_tumor_depth | Depth of coverage from specific tumor sample | -| proband_sample_id_tumor_alt_depth | Allele count at site for proband for specific tumor sample| -| proband_sample_id_tumor_ref_depth | Reference count at site for proband for specific tumor sample | +| proband_sample_id_tumor_alt_depth | Allele count at site from specific proband tumor sample| +| proband_sample_id_tumor_ref_depth | Reference count at site from specific proband tumor sample | + +![WF Visualized](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/loh.png) ### Running it locally on a laptop? 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zI0F$I_&6mN?Arh9hnK()|Nn3P7q$KWRr-J7gan*kSdLN2)UR#s!2ti1Z)(Vw%bExN EAK72tK>z>% literal 0 HcmV?d00001 From d724045e89838cc043520d673574d5278d64c319 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 12:46:20 -0400 Subject: [PATCH 04/18] correct the path --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 26f74dd..7a69007 100644 --- a/README.md +++ b/README.md @@ -79,7 +79,7 @@ LOH workflow will generate a tab-separated values file with following headers: | proband_sample_id_tumor_alt_depth | Allele count at site from specific proband tumor sample| | proband_sample_id_tumor_ref_depth | Reference count at site from specific proband tumor sample | -![WF Visualized](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/loh.png) +![WF Visualized](https://github.com/d3b-center/tumor-loh-app-dev/tree/feature/public-release/docs/logo/loh.png) ### Running it locally on a laptop? From fe86aa26857217ecc195bb0857f60a64eb0e0d8a Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 12:52:53 -0400 Subject: [PATCH 05/18] test images --- README.md | 2 +- docs/logo/loh.png | Bin 105063 -> 0 bytes docs/logo/loh_wf.svg | 605 +++++++++++++++++++++++++++++++++++++++++++ 3 files changed, 606 insertions(+), 1 deletion(-) delete mode 100644 docs/logo/loh.png create mode 100644 docs/logo/loh_wf.svg diff --git a/README.md b/README.md index 7a69007..582ad5b 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,6 @@ # Kids First Loss of Heterozygosity (LOH) -![data service logo](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/kfdrc-logo-sm.png) +![data service logo](https://github.com/d3b-center/tumor-loh-app-dev/tree/feature/public-release/docs/logo/kids_first_logo.png) The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. 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zI0F$I_&6mN?Arh9hnK()|Nn3P7q$KWRr-J7gan*kSdLN2)UR#s!2ti1Z)(Vw%bExN EAK72tK>z>% diff --git a/docs/logo/loh_wf.svg b/docs/logo/loh_wf.svg new file mode 100644 index 0000000..bf2a4c4 --- /dev/null +++ b/docs/logo/loh_wf.svg @@ -0,0 +1,605 @@ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + tool2 + + + run_germline + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + tool2 + + + run_tumor_tool + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + type_input + + + BS_ID + + + + + + + + + + + + + + + + + + type_input + + + participant_id + + + + + + + + + + + + + + + + + + type_input + + + frequency + + + + + + + + + + + + + + + + + + file_input + + + peddy_file + + + + + + + + + + + + + + + + + + file_input + + + bamscrams + + + + + + + + + + + + + + + + + + type_input + + + minDepth + + + + + + + + + + + + + + + + + + file_input + + + reference + + + + + + + + + + + + + + + + + + file_input + + + sample_vcf_file + + + + + + + + + + + + + + + + + + type_input + + + bamcramsampleIDs + + + + + + + + + + + + + + + + + + type_input + + + ram_germline + + + + + + + + + + + + + + + + + + type_input + + + ram_tumor + + + + + + + + + + + + + + + + + + type_input + + + minCore + + + + + + + + + + + + + + + + + + file_output + + + output_file + + + + + + + + + + + + + \ No newline at end of file From cb753964409df8549e515dd691ef5a37c6c9be78 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 12:54:58 -0400 Subject: [PATCH 06/18] correct the images --- README.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/README.md b/README.md index 582ad5b..0c4a227 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,6 @@ # Kids First Loss of Heterozygosity (LOH) -![data service logo](https://github.com/d3b-center/tumor-loh-app-dev/tree/feature/public-release/docs/logo/kids_first_logo.png) +![data service logo](https://github.com/d3b-center/tumor-loh-app-dev/tree/feature/public-release/docs/logo/kids_first_logo.svg) The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This workflow is designed to analyze LOH for family trios as well as multiple proband tumor samples. @@ -79,7 +79,7 @@ LOH workflow will generate a tab-separated values file with following headers: | proband_sample_id_tumor_alt_depth | Allele count at site from specific proband tumor sample| | proband_sample_id_tumor_ref_depth | Reference count at site from specific proband tumor sample | -![WF Visualized](https://github.com/d3b-center/tumor-loh-app-dev/tree/feature/public-release/docs/logo/loh.png) +![WF Visualized](https://github.com/d3b-center/tumor-loh-app-dev/tree/feature/public-release/docs/logo/loh_wf.svg) ### Running it locally on a laptop? From eec3375e38e0454466fa39ec700c4a8e5ead0038 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 12:59:40 -0400 Subject: [PATCH 07/18] images --- README.md | 3 +-- docs/logo/loh.png | Bin 0 -> 105063 bytes 2 files changed, 1 insertion(+), 2 deletions(-) create mode 100644 docs/logo/loh.png diff --git a/README.md b/README.md index 0c4a227..3325bbe 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,6 @@ # Kids First Loss of Heterozygosity (LOH) -![data service logo](https://github.com/d3b-center/tumor-loh-app-dev/tree/feature/public-release/docs/logo/kids_first_logo.svg) +![data service logo](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/kfdrc-logo-sm.png) The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This workflow is designed to analyze LOH for family trios as well as multiple proband tumor samples. @@ -79,7 +79,6 @@ LOH workflow will generate a tab-separated values file with following headers: | proband_sample_id_tumor_alt_depth | Allele count at site from specific proband tumor sample| | proband_sample_id_tumor_ref_depth | Reference count at site from specific proband tumor sample | -![WF Visualized](https://github.com/d3b-center/tumor-loh-app-dev/tree/feature/public-release/docs/logo/loh_wf.svg) ### Running it locally on a laptop? diff --git a/docs/logo/loh.png b/docs/logo/loh.png new file mode 100644 index 0000000000000000000000000000000000000000..1ac7970d75f068a8ac01153e15ef6b63b14d7e90 GIT binary patch literal 105063 zcmb5VWmp_r(=`l%;10pv3GVJ5+}(o*cbA~SEkJMx7Tg_zLvRW1?(X_F_lcbIe80Y2 zGZ&Md*}c1K*REP?RduMmtT;RjHVg;|2)v|(h$09G7(58bt63-r;K(KCkQE3BOo_R$ zu)L(OFrmD?jfwdOV-OID(1b)t<+xcC-{a?ecFk9>6oueLqh&#;h^pm<>Dh!(2;(uq zK)Zwb)?i9sBO}YB7l-&;g1#?NFE59BJEIdVDxiY_%ci`9i0j{U+r?*7S>{84fOMBz?cPgkTu@-Rq_j>e|+X0`KRZbe66` z%sQ58llm@?N1lr)VFR!{K?vzOp#9;zxjHD2@K&Y1CWB;N5qp0;knRW96HOu)#TL04 z1E-KV?E|NfX2RyhLPhQoQ^y10N$!m!2NhrwdAOkQMu`nR2rN%fCqy9$_k^_G-IW4| zB60GAMj;V&CFW;dZQ_yi8Av7^>0c-7ce9|6^S?$hmKb*(cpjNPZ6o8AVbkfvRIsm{ zq=&1J&>L-;Bm=carvML@u99HD;_vTgYU?xKGA&=v)A)W46I~_)i;%h{5`!q7P7#O7 zIMe}md}98Y4mxi7U3{g+#9-X;WJ0dkkH>zYNFE1QEEOL9}O@Md=%Ym)=QM2H#BGd7h@b^Qb#dO4Xl`M5!ysaP2&X5)0hyQY(%@Q` z@__yR8XM&86f+-$y^G`nq=nD^MH|U%s2$f@*`Z3sCGpAPUlB zg3y15az*SRgz6AP3xP2gXpBaFk1#7B-3Nab^eT&i68%$fEFmN5M^&NyOyUB}x53}C zGTSfH>MU<$#I-VYM!k0=SO{9sTCiJ?bt3sftVQ&Mmc_2K#7CDI(8AGsaM#0E^$i&$ zlRij;=Al&)IG{LSO+;XQZ|*g$uuzAXLf#LK=+4tu(`PqyF~BL`uF#qBKA=Php7@@; zu5Cxeflr63<~twC9nKxx6o~WPt1IrL(G{l|kq?2sXM0uZH2B2ywC?2N4WmCwd&r73 z6cH5E0CWnp2Q+O!N&sjFrb2QbVIvAj2vt{27n33HDbp$PDQ&38I0a{nkR-7pU17Lm zyko+o_??I-Cex3qymjCro|Ym~|HGVz)}M1P?E5P$v6Klhcm zU}m{kmQa?$>gNt>QUB~1Vkx^A!$`xzHM{PVPxJ{;?|R;SAWI*&$c4#OdgqcLmQeU( z>Icz}-5)p!h*Ww?_=@d?m_@EdP|9a2R|VS2yZPA#5Xx)`aysIRQZ3SWnFi)?*ukHL zsI&31oJPfzRg@2vbCeScRSVJ;4poE;v5M>_$0yl}@Rary?s=p<&@DgPP6cUPR#^|- zW%6k)3hNZ;7A$2hl{{&DEL|;$`BtHlBe`Bdt~nvitjw(CP`70~lvrh6rDJk#iZRT` zcJGH?Y@GKduSUo+O0{XKpf<9$tX9M$|4Q^Qk;{S$pNoo%jLXz1$f>Z=x{-JhWznH= zWcQmRI&XxoSeppJ*u?nxMBJ{*zSB;`F7pM+<@p891py8cd=f+`1Q|pL1W8b1kYErs z{5~83CUSU^Sicx|_*^(2CUbaB#54{&79!&$7OogVPULA{fZ>{T<>c3%jYz7gc=In7 zjRVyaaBNlVQfx&GP38zG+(}`BF3B!K?Spn-NYa&E0~U%7Mft0C7aH^ zzqKe`U8rx++O8dXG%>bGFsYf|%c}WQ)8>@!6n2S&!;I4^6F7vY^{FK45PRNae%g7< zS*U5z`TC&nO7Lp`Q0uUbi-Su7Zx1gB4<2tFj~efo$y}Pwf#m!fYES=!dBlWyzT7av zaPn|(>a%@?t)vs@1`|sZqa26LakICYms+}Ijb$0mBJI+lX`QYMuS>c|#Jv(C7@}XV zca-2~D(XO|q{|-@Ka}>|#?Qg#lUqJ{Ex~SX?yKzlTrF9{+toRG>=Iw5=+7TE?V4V4 z2|RY=57+ZuR&8s-wL-{H8o)YrxzOA6q4LE=!k5QCVHu^7$DTm}hrETff^|pxfJTF0 z4FgJ)8f`@s)c|*-cvO}(BX4acx>LolU!GuKwK{pyy&BxHEbc1yA<`Ao9EB?ELxf_u zCXx!yCD9ypBS#~x`#kr@?D&Cqk~Q&12xQLdG3X8AHqMK13%-@lZj5r5%Q=`q`F- zc5CvnmgBWwSKnW6jBg|o*A{4BaCOt?zTFhIkl6E^JgwfT4s!H#_q?O3_{5>&X&yQp zYb@1Bx)u9M@>p>@cRn#v0_^KOb7puT)=)Cbdk%}ly0%k+HB4xconG^eNz)ILRGCRWmJ!AGa;O2zh2Q?o13g@ z+tk>6v)!VF(NapL$IZ{)_6)g9um^(%^YyjMbW?HJ9r9z&PH!Z9EsnGGjVdK&v+jEp zWz7p?5})xFA`>S)S~-SSt5X}R0tDH2`OTvb_=EmW)v9?GT4_ou9*7B5xiR76!W ze9#{~%jW7DD|+hl91)%6rqWG4a;|C*jgE>I%Np9uo~Oaqz*-In|QA(5SZH9%FojK=i(PkN>?0#A)M~=J6xyIV;L)H4W{cuEjVycfD(XNos z{@PS9h7sLT1Dbo`y8ZXfA~nxa?GHvZX-#|Xmlv$tmieX3W$cT#^H(@+Kw1hII+w)%3d5 zDW3ao!|U$80x^()l@I@+>tXVW^X_~_{R(3tv9c^rPpkE{rw24%c5fK*|vrQP=g7AVCsBn1)H1p|Z6D zNO(X#SW-{#-n!VUW+3Huf+$;R+2QdVZzF8_`}9rXKI<$$R|}+H+Fa?q4|*Mh^(}TD zz^|&t>XIfhG9Yh(&rl$s!R8>~z$Z}PjSaj3s2dvu0tx&@0p22+uYP|E2A>J``}3>W zmmdl$2}?=>f0c~vjg74x%xoMNAG)b8v7$40Z)0do=W1p9G6aOj zl?(W2W$b7`=xX)B+JVcJm-v6b-~v9s9Hu8G{NGO;EqRI6W#kEkZS0K++2|PP7>M~` z2nh*!?BAPkDT;{w8V>xAm)OkF(Uyyz-o?d*&V`xI#@>{kk&}~?o`H#;iHR2Y1+9ad zwWEP6t+fNmKPUNp9uZ>)BYSgOM{^r%!k67M0%Qy6lbPV)=&J7IZc{$1@Z|-XRK|{pc3NQ~a2Om2VC(r+W|Nr^s?~MN%ss68# z9E|@S`LA#O??@E~V|!s6D_}}TzP~H;Yw*9n{56n={^id9Wr=^x{J*1so%vvR=>M!4 zA58G-?hy!x0EncBpt39IK?`&>uDIK^4%1Yt&F6^shxesju^;-w-p^Lj&L-)=?L3AV@`8!F81+1eZOF#bl5gOw

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zI0F$I_&6mN?Arh9hnK()|Nn3P7q$KWRr-J7gan*kSdLN2)UR#s!2ti1Z)(Vw%bExN EAK72tK>z>% literal 0 HcmV?d00001 From b74c7311184b8ff9a6eb2ad09978454488968041 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 14:55:05 -0400 Subject: [PATCH 08/18] improve the doc --- README.md | 4 +-- workflow/run_LOH_app.cwl | 60 ++++++++++++++++++++++++++++++++++++++-- 2 files changed, 60 insertions(+), 4 deletions(-) diff --git a/README.md b/README.md index 3325bbe..019a6ec 100644 --- a/README.md +++ b/README.md @@ -8,7 +8,7 @@ Moreover, this workflow has been deployed on [cavatica](https://cavatica.sbgenom ### Application Description -The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and tumor tool. +The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and Tumor tool. #### Docker Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH @@ -47,7 +47,7 @@ Tumor tool LOH application will output a tab-separated values file mapped data from germline tool and tumor tool. ``` -output_file: { type: File, doc: output file from LOH app, outputSource: run_tumor_tool/loh_output_file_tool } +output_file: { type: File, doc: A tsv file with gathered data from germline and tumor tool } ``` #### Output headers diff --git a/workflow/run_LOH_app.cwl b/workflow/run_LOH_app.cwl index 4c294f9..c4ef1ab 100644 --- a/workflow/run_LOH_app.cwl +++ b/workflow/run_LOH_app.cwl @@ -3,7 +3,57 @@ cwlVersion: v1.2 class: Workflow id: run_LOH_app -doc: This CWL workflow assesses the loss of heterozygosity (LOH) in the tumor for rare germline variants. Order of operations- This workflow runs bcftools to extract data and prepare list of locations from vcf and feed it into bam-readcount to compute VAF and later parse, merge germline and tumor data together. Github- https://github.com/d3b-center/tumor-loh-app-dev +label: Kids First Loss of Heterozygosity +doc: | + # Kids First Loss of Heterozygosity (LOH) + + ![data service logo](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/kfdrc-logo-sm.png) + + The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This workflow is designed to analyze LOH for family trios as well as multiple proband tumor samples. + + ### Description + + The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and Tumor tool. + + #### Docker + Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH + + #### Germline Tool + + Germline tool filters germline annotations to retain variants based on gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. It requires vcf file, proband sample id, ram as required inputs and peddy file as optional input which is required for family trios. It outputs variant information such as gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, variant allele frequency and list of coordinates that will be an input to tumor tool. + + #### Tumor Tool + Tumor tool search in paired proband tumor sample for aligned reads in the regions where rare variants from the germline tool exists and exact allele/reference count, allele/reference depth and calculate the variant allele frequency VAF. Tumor tool have the capability to search multiple tumor samples for proband and if applicable, parental and maternal tumor samples. To exact reads from the bam/cram files, this tool utilizes [bam-readcount](https://github.com/genome/bam-readcount) and wraps it with python script to shape the output in a tabular format. + + ### LOH Inputs + ``` + Germline tool + # Required + BS_ID: { doc: provide BS id for germline normal,type: string } + frequency: { doc: provide popmax cutoff for rare germline variants, type: 'float?', default: 0.01 } + # Optional + ram_germline: { doc: Provide ram (in GB) based on the size of vcf,type: 'int?', default: 8} + # Required for family trios otherwise not required + peddy_file: { doc: provide ped file for the trio, type: 'File?' } + Tumor tool + # Required + participant_id: { doc: provide participant id for this run, type: string } + bamscrams: { doc: tumor input file in cram or bam format with their index file, type: 'File[]' , secondaryFiles: [ { pattern: ".crai", required: false }, { pattern: ".bai", required: false } ] } + reference: { doc: human reference in fasta format with index file, type: File,secondaryFiles: [ .fai ] } + sample_vcf_file: { doc: provide germline vcf file for this sample, type: File } + # Optional + minDepth: { doc: provide minDepth to consider for tumor reads, type: 'int?', default: 1 } + bamcramsampleIDs: { doc: provide unique identifers (in the same order) for cram/bam files provided under bamcrams tag. Default is sample ID pulled from bam/cram files., type: 'string[]?' } + ram_tumor: { doc: Provide ram (in GB) for tumor tool based on the number cram/bam inputs, type: 'int?', default: 16} + minCore: { type: 'int?', default: 16, doc: "Minimum number of cores for tumor tool based on the number cram/bam inputs" } + ``` + + ### LOH Output + + LOH application will output a tab-separated values file mapped data from germline tool and tumor tool. + ``` + output_file: { type: File, doc: A tsv file with gathered data from germline and tumor tool} + ``` requirements: - class: StepInputExpressionRequirement @@ -49,4 +99,10 @@ steps: ram: ram_tumor minCore: minCore out: - [ loh_output_file_tool,log_output ] \ No newline at end of file + [ loh_output_file_tool,log_output ] + +"sbg:license": Apache License 2.0 +"sbg:publisher": KFDRC +"sbg:links": +- id: 'https://github.com/d3b-center/tumor-loh-app-dev/releases/tag/v1.0.1' + label: github-release \ No newline at end of file From 0e88465f11a45a5eadb72d38f2e2efd20c9a16de Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 15:00:13 -0400 Subject: [PATCH 09/18] del the file --- docs/logo/loh_wf.svg | 605 ------------------------------------------- 1 file changed, 605 deletions(-) delete mode 100644 docs/logo/loh_wf.svg diff --git a/docs/logo/loh_wf.svg b/docs/logo/loh_wf.svg deleted file mode 100644 index bf2a4c4..0000000 --- a/docs/logo/loh_wf.svg +++ /dev/null @@ -1,605 +0,0 @@ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - tool2 - - - run_germline - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - tool2 - - - run_tumor_tool - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - type_input - - - BS_ID - - - - - - - - - - - - - - - - - - type_input - - - participant_id - - - - - - - - - - - - - - - - - - type_input - - - frequency - - - - - - - - - - - - - - - - - - file_input - - - peddy_file - - - - - - - - - - - - - - - - - - file_input - - - bamscrams - - - - - - - - - - - - - - - - - - type_input - - - minDepth - - - - - - - - - - - - - - - - - - file_input - - - reference - - - - - - - - - - - - - - - - - - file_input - - - sample_vcf_file - - - - - - - - - - - - - - - - - - type_input - - - bamcramsampleIDs - - - - - - - - - - - - - - - - - - type_input - - - ram_germline - - - - - - - - - - - - - - - - - - type_input - - - ram_tumor - - - - - - - - - - - - - - - - - - type_input - - - minCore - - - - - - - - - - - - - - - - - - file_output - - - output_file - - - - - - - - - - - - - \ No newline at end of file From 7576c74dd16482ce7b1f25ab757b0e65588163e3 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 15:09:36 -0400 Subject: [PATCH 10/18] add category --- workflow/run_LOH_app.cwl | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/workflow/run_LOH_app.cwl b/workflow/run_LOH_app.cwl index c4ef1ab..f2bba2b 100644 --- a/workflow/run_LOH_app.cwl +++ b/workflow/run_LOH_app.cwl @@ -103,6 +103,11 @@ steps: "sbg:license": Apache License 2.0 "sbg:publisher": KFDRC +"sbg:categories": +- DNA +- WGS +- WXS +- GVCF "sbg:links": - id: 'https://github.com/d3b-center/tumor-loh-app-dev/releases/tag/v1.0.1' label: github-release \ No newline at end of file From fd2420b0a6bcc68e89cbb3ee87d20f2aa23805bb Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Mon, 26 Jun 2023 16:24:13 -0400 Subject: [PATCH 11/18] add LOH doc file --- docs/LOH_readme.md | 25 +++++++++++++++++++++++++ 1 file changed, 25 insertions(+) create mode 100644 docs/LOH_readme.md diff --git a/docs/LOH_readme.md b/docs/LOH_readme.md new file mode 100644 index 0000000..5a362f2 --- /dev/null +++ b/docs/LOH_readme.md @@ -0,0 +1,25 @@ +# Kids First Loss of Heterozygosity (LOH) + +![data service logo](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/kfdrc-logo-sm.png) + +LOH workflow assesses the loss of heterozygosity (LOH) in the tumor for rare germline variants. +Order of operations: This workflow runs bcftools to extract data and prepare list of locations from vcf provided and feed it into bam-readcount to compute VAF and later parse, merge germline and tumor data together. + +## Inputs + +- BS_ID : Sample id for proband +- frequency: cut off for rare germline variants based on gnomad_3_1_1_AF_popmax tag +- ram_germline : provide ram for germline tool which is directly related with size of VCF file provide in sample_vcf_file +- peddy_file: details about the family trio +- participant_id: provide participant_id for proband. It is used just to name the output file. +- bamscrams: provide multiple bam/cram files for proband and family trios +- reference: human reference file in fasta format +- sample_vcf_file: vcf file to exact germline calls +- minDepth: minimum depth of the reads that should be considered in the tunor analysis +- bamcramsampleIDs: provide bam/cram sampleids in the same order as provided in bamscrams. +- ram_tumor: ram for tumor tool, which is strongly connected with the size of the cram files and number of cram files provided +- mincore: provide number of processor. Each cram will be split into 32 parts for multiprocessing and results will be merge back. High number of processors are recommended. + +## output + +- output_file: a tsv file with mapped variant data from germline and tumor tool containing germline VAF and tumor VAF From d3f24536b489b5de6e12a4918c0e70a123726992 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Tue, 27 Jun 2023 09:50:52 -0400 Subject: [PATCH 12/18] update the doc --- README.md | 27 +++++++++++++-------------- workflow/run_LOH_app.cwl | 2 +- 2 files changed, 14 insertions(+), 15 deletions(-) diff --git a/README.md b/README.md index 019a6ec..4b5290c 100644 --- a/README.md +++ b/README.md @@ -4,8 +4,6 @@ The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This workflow is designed to analyze LOH for family trios as well as multiple proband tumor samples. -Moreover, this workflow has been deployed on [cavatica](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/apps/Loss_of_Heterozygosity). It was tested with a [proband only run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/522d128a-2195-4c9c-8339-1709da16821d/) and a [trio run](https://cavatica.sbgenomics.com/u/d3b-bixu/tumor-loh-dev/tasks/d7f6b667-35ef-46a7-a666-970a78ef3175/) - ### Application Description The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and Tumor tool. @@ -18,36 +16,37 @@ Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH Germline tool filters germline annotations to retain variants based on gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. It requires vcf file, proband sample id, ram as required inputs and peddy file as optional input which is required for family trios. It outputs variant information such as gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, variant allele frequency and list of coordinates that will be an input to tumor tool. #### Tumor Tool -Tumor tool search in paired proband tumor sample for aligned reads in the regions where rare variants from the germline tool exists and exact allele/reference count, allele/reference depth and calculate the variant allele frequency VAF. Tumor tool have the capability to search multiple tumor samples for proband and if applicable, parental and maternal tumor samples. To exact reads from the bam/cram files, this tool utilizes [bam-readcount](https://github.com/genome/bam-readcount) and wraps it with python script to shape the output in a tabular format. +Tumor tool search in paired proband tumor sample for aligned reads in the regions where rare variants from the germline tool exists and exact allele/reference count, allele/reference depth and calculate variant allele frequency(VAF). Tumor tool have the capability to search in multiple tumor samples for proband and if applicable, parental and maternal tumor samples. To exact reads from the bam/cram files, this tool utilizes [bam-readcount](https://github.com/genome/bam-readcount) and wraps it with python script to shape the output in a tabular format. ### LOH Inputs ``` Germline tool # Required - BS_ID: { doc: provide BS id for germline normal,type: string } - frequency: { doc: provide popmax cutoff for rare germline variants, type: 'float?', default: 0.01 } + BS_ID:{ doc: provide BS id for germline normal,type: string } + frequency:{ doc: provide popmax cutoff for rare germline variants, type: 'float?', default: 0.01 } # Optional - ram_germline: { doc: Provide ram (in GB) based on the size of vcf,type: 'int?', default: 8} + ram_germline:{ doc: Provide ram (in GB) based on the size of vcf,type: 'int?', default: 8} # Required for family trios otherwise not required - peddy_file: { doc: provide ped file for the trio, type: 'File?' } + peddy_file:{ doc: provide ped file for the trio, type: 'File?' } + Tumor tool # Required - participant_id: { doc: provide participant id for this run, type: string } - bamscrams: { doc: tumor input file in cram or bam format with their index file, type: 'File[]' , secondaryFiles: [ { pattern: ".crai", required: false }, { pattern: ".bai", required: false } ] } - reference: { doc: human reference in fasta format with index file, type: File,secondaryFiles: [ .fai ] } + participant_id:{ doc: provide participant id for this run, type: string } + bamscrams:{ doc: tumor input file in cram or bam format with their index file, type: 'File[]' , secondaryFiles: [ { pattern: ".crai", required: false }, { pattern: ".bai", required: false } ] } + reference:{ doc: human reference in fasta format with index file, type: File,secondaryFiles: [ .fai ] } sample_vcf_file: { doc: provide germline vcf file for this sample, type: File } # Optional - minDepth: { doc: provide minDepth to consider for tumor reads, type: 'int?', default: 1 } + minDepth:{ doc: provide minDepth to consider for tumor reads, type: 'int?', default: 1 } bamcramsampleIDs: { doc: provide unique identifers (in the same order) for cram/bam files provided under bamcrams tag. Default is sample ID pulled from bam/cram files., type: 'string[]?' } - ram_tumor: { doc: Provide ram (in GB) for tumor tool based on the number cram/bam inputs, type: 'int?', default: 16} - minCore: { type: 'int?', default: 16, doc: "Minimum number of cores for tumor tool based on the number cram/bam inputs" } + ram_tumor:{ doc: Provide ram (in GB) for tumor tool based on the number cram/bam inputs, type: 'int?', default: 16} + minCore:{ type: 'int?', default: 16, doc: "Minimum number of cores for tumor tool based on the number cram/bam inputs" } ``` ### LOH Output LOH application will output a tab-separated values file mapped data from germline tool and tumor tool. ``` -output_file: { type: File, doc: A tsv file with gathered data from germline and tumor tool } +output_file:{ type: File, doc: A tsv file with gathered data from germline and tumor tool } ``` #### Output headers diff --git a/workflow/run_LOH_app.cwl b/workflow/run_LOH_app.cwl index f2bba2b..f2b5bd6 100644 --- a/workflow/run_LOH_app.cwl +++ b/workflow/run_LOH_app.cwl @@ -39,7 +39,7 @@ doc: | # Required participant_id: { doc: provide participant id for this run, type: string } bamscrams: { doc: tumor input file in cram or bam format with their index file, type: 'File[]' , secondaryFiles: [ { pattern: ".crai", required: false }, { pattern: ".bai", required: false } ] } - reference: { doc: human reference in fasta format with index file, type: File,secondaryFiles: [ .fai ] } + reference: { doc: human reference in fasta format with index file, type: File,secondaryFiles: [ .fai ],"sbg:suggestedValue": {class: File, path: 60639014357c3a53540ca7a3, name: Homo_sapiens_assembly38.fasta} } sample_vcf_file: { doc: provide germline vcf file for this sample, type: File } # Optional minDepth: { doc: provide minDepth to consider for tumor reads, type: 'int?', default: 1 } From e98122330df3ad746eeb3d0fae7621365abb6cde Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Tue, 27 Jun 2023 09:55:06 -0400 Subject: [PATCH 13/18] Test image --- README.md | 3 +++ 1 file changed, 3 insertions(+) diff --git a/README.md b/README.md index 4b5290c..d2fc583 100644 --- a/README.md +++ b/README.md @@ -41,6 +41,9 @@ Tumor tool ram_tumor:{ doc: Provide ram (in GB) for tumor tool based on the number cram/bam inputs, type: 'int?', default: 16} minCore:{ type: 'int?', default: 16, doc: "Minimum number of cores for tumor tool based on the number cram/bam inputs" } ``` +### LOH schematic + +![LOH schematic](https://github.com/d3b-center/tumor-loh-app-dev/blob/master/docs/logo/chop_logo.svg) ### LOH Output From 31a16862c28e42b8d44dad11e84a0d7f22e12972 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Tue, 27 Jun 2023 11:10:38 -0400 Subject: [PATCH 14/18] Add screenshots --- README.md | 18 +++++++++++------- docs/{LOH_readme.md => README.md} | 10 +++++++++- docs/logo/family_trio_run.png | Bin 0 -> 177367 bytes docs/logo/proband_run.png | Bin 0 -> 169109 bytes workflow/run_LOH_app.cwl | 2 +- 5 files changed, 21 insertions(+), 9 deletions(-) rename docs/{LOH_readme.md => README.md} (84%) create mode 100644 docs/logo/family_trio_run.png create mode 100644 docs/logo/proband_run.png diff --git a/README.md b/README.md index d2fc583..118ef36 100644 --- a/README.md +++ b/README.md @@ -8,8 +8,11 @@ The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and Tumor tool. -#### Docker -Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH +#### Basic info +- Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH/1.0.1 +- tested with + - Seven Bridges Cavatica Platform: https://cavatica.sbgenomics.com/ + - cwltool: https://github.com/common-workflow-language/cwltool/releases/tag/3.1.20221201130942 #### Germline Tool @@ -19,13 +22,13 @@ Germline tool filters germline annotations to retain variants based on gnomad_3_ Tumor tool search in paired proband tumor sample for aligned reads in the regions where rare variants from the germline tool exists and exact allele/reference count, allele/reference depth and calculate variant allele frequency(VAF). Tumor tool have the capability to search in multiple tumor samples for proband and if applicable, parental and maternal tumor samples. To exact reads from the bam/cram files, this tool utilizes [bam-readcount](https://github.com/genome/bam-readcount) and wraps it with python script to shape the output in a tabular format. ### LOH Inputs -``` +```yaml Germline tool # Required - BS_ID:{ doc: provide BS id for germline normal,type: string } + BS_ID:{ doc: provide BS id for germline normal, type: string } frequency:{ doc: provide popmax cutoff for rare germline variants, type: 'float?', default: 0.01 } # Optional - ram_germline:{ doc: Provide ram (in GB) based on the size of vcf,type: 'int?', default: 8} + ram_germline:{ doc: Provide ram (in GB) based on the size of vcf, type: 'int?', default: 8} # Required for family trios otherwise not required peddy_file:{ doc: provide ped file for the trio, type: 'File?' } @@ -43,12 +46,12 @@ Tumor tool ``` ### LOH schematic -![LOH schematic](https://github.com/d3b-center/tumor-loh-app-dev/blob/master/docs/logo/chop_logo.svg) +![LOH schematic](https://github.com/d3b-center/tumor-loh-app-dev/blob/master/docs/logo/loh.png) ### LOH Output LOH application will output a tab-separated values file mapped data from germline tool and tumor tool. -``` +```yaml output_file:{ type: File, doc: A tsv file with gathered data from germline and tumor tool } ``` @@ -81,6 +84,7 @@ LOH workflow will generate a tab-separated values file with following headers: | proband_sample_id_tumor_alt_depth | Allele count at site from specific proband tumor sample| | proband_sample_id_tumor_ref_depth | Reference count at site from specific proband tumor sample | +More detailed can be found [here](https://github.com/d3b-center/tumor-loh-app-dev/tree/master/docs/README.md) ### Running it locally on a laptop? diff --git a/docs/LOH_readme.md b/docs/README.md similarity index 84% rename from docs/LOH_readme.md rename to docs/README.md index 5a362f2..0dad64f 100644 --- a/docs/LOH_readme.md +++ b/docs/README.md @@ -20,6 +20,14 @@ Order of operations: This workflow runs bcftools to extract data and prepare lis - ram_tumor: ram for tumor tool, which is strongly connected with the size of the cram files and number of cram files provided - mincore: provide number of processor. Each cram will be split into 32 parts for multiprocessing and results will be merge back. High number of processors are recommended. -## output +## Output - output_file: a tsv file with mapped variant data from germline and tumor tool containing germline VAF and tumor VAF + +## Demo Proband-only Cavatica Task + +![LOH schematic](https://github.com/d3b-center/tumor-loh-app-dev/blob/master/docs/logo/proband_run.png) + +## Demo Family-trio Cavatica Task + +![LOH schematic](https://github.com/d3b-center/tumor-loh-app-dev/blob/master/docs/logo/proband_run.png) \ No newline at end of file diff --git a/docs/logo/family_trio_run.png b/docs/logo/family_trio_run.png new file mode 100644 index 0000000000000000000000000000000000000000..d011071cb7cc03cce319f8ef2199e456c5fa6d85 GIT binary patch literal 177367 zcmeFZgMVMk);OFrY0%hB(%80bG`7>&wryLD+1R%Gjcwa#%r`yf9zOT>_Yb_u&S%e_ z#aXikd)C?sk&zOC`GD~O2nYy9OjJ+~2ngH`2na+6;@w+~?J*e7+YQ)GPJ|z*eDu@7 zTbbHGMa)oA5{Uc_009I7j0gns6Xop}2$&w|Cl&JBAAOJs5a@69KtRH8cOZINdIok{ zW_EfOTsm5Irnid?2naeF88wQn~i>1Ae 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WOYYjZ0=vI}Ke_u#QU&*(zWRSPT Date: Tue, 27 Jun 2023 11:12:55 -0400 Subject: [PATCH 15/18] fix readme --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 118ef36..d611d7f 100644 --- a/README.md +++ b/README.md @@ -84,7 +84,7 @@ LOH workflow will generate a tab-separated values file with following headers: | proband_sample_id_tumor_alt_depth | Allele count at site from specific proband tumor sample| | proband_sample_id_tumor_ref_depth | Reference count at site from specific proband tumor sample | -More detailed can be found [here](https://github.com/d3b-center/tumor-loh-app-dev/tree/master/docs/README.md) +More information can be found [here](https://github.com/d3b-center/tumor-loh-app-dev/tree/master/docs/README.md) ### Running it locally on a laptop? From c516aad816322b039153dca235328945b87f524d Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Wed, 28 Jun 2023 15:27:24 -0400 Subject: [PATCH 16/18] Update README.md Co-authored-by: Alex Sickler --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index d611d7f..1572193 100644 --- a/README.md +++ b/README.md @@ -19,7 +19,7 @@ The Kids First Loss of Heterozygosity application is divided into two tools: Ger Germline tool filters germline annotations to retain variants based on gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. It requires vcf file, proband sample id, ram as required inputs and peddy file as optional input which is required for family trios. It outputs variant information such as gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, variant allele frequency and list of coordinates that will be an input to tumor tool. #### Tumor Tool -Tumor tool search in paired proband tumor sample for aligned reads in the regions where rare variants from the germline tool exists and exact allele/reference count, allele/reference depth and calculate variant allele frequency(VAF). Tumor tool have the capability to search in multiple tumor samples for proband and if applicable, parental and maternal tumor samples. To exact reads from the bam/cram files, this tool utilizes [bam-readcount](https://github.com/genome/bam-readcount) and wraps it with python script to shape the output in a tabular format. +Search in paired proband tumor sample for aligned reads in the regions where rare variants from the germline exist and exact allele/reference count, allele/reference depth, and calculate variant allele frequency(VAF). Tumor tool has the capability to search in multiple tumor samples for proband and if applicable, paternal and maternal tumor samples. To extract reads from the bam/cram files, this tool utilizes [bam-readcount](https://github.com/genome/bam-readcount) and wraps it with a python script to shape the output in a tabular format. ### LOH Inputs ```yaml From ab5546c5eb0d259665be993811065dc36e669b9c Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Thu, 29 Jun 2023 10:59:50 -0400 Subject: [PATCH 17/18] Address PR feedback --- README.md | 14 +++++++------- docs/README.md | 2 +- tools/run_gene_extract_list_prepare.cwl | 2 +- tools/run_readcount_parser.cwl | 1 + workflow/run_LOH_app.cwl | 19 ++++++++++++------- 5 files changed, 22 insertions(+), 16 deletions(-) diff --git a/README.md b/README.md index 1572193..f770dcb 100644 --- a/README.md +++ b/README.md @@ -2,11 +2,7 @@ ![data service logo](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/kfdrc-logo-sm.png) -The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This workflow is designed to analyze LOH for family trios as well as multiple proband tumor samples. - -### Application Description - -The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and Tumor tool. +The Kids First Loss of Heterozygosity Preprocessing (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This preprocessing is designed to compute variant allele frequency (VAF) for multiple proband tumor samples and can also map germline VAF for family trios if trio germline VCF file is provided. #### Basic info - Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH/1.0.1 @@ -14,6 +10,10 @@ The Kids First Loss of Heterozygosity application is divided into two tools: Ger - Seven Bridges Cavatica Platform: https://cavatica.sbgenomics.com/ - cwltool: https://github.com/common-workflow-language/cwltool/releases/tag/3.1.20221201130942 +### Application Description + +The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and Tumor tool. + #### Germline Tool Germline tool filters germline annotations to retain variants based on gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. It requires vcf file, proband sample id, ram as required inputs and peddy file as optional input which is required for family trios. It outputs variant information such as gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, variant allele frequency and list of coordinates that will be an input to tumor tool. @@ -57,7 +57,7 @@ output_file:{ type: File, doc: A tsv file with gathered data from germline and t #### Output headers -LOH workflow will generate a tab-separated values file with following headers: +Preprocessing LOH will generate a tab-separated values file with following headers: | Headers | Description | |:-------:|:--------:| | BS_ID | Sample Id for germline sample | @@ -88,7 +88,7 @@ More information can be found [here](https://github.com/d3b-center/tumor-loh-app ### Running it locally on a laptop? -It is recommended to run this workflow on a system with a high number of CPUs and memory (>=16 GB). The basic requirement is a running docker engine and CWL tools. Command line to run the LOH workflow locally is: +It is recommended to run this CWL workflow on a system with a high number of CPUs and memory (>=16 GB). The basic requirement is a running docker engine and CWL tools. Command line to run the LOH workflow locally is: ``` cwltool workflow/run_LOH_app.cwl sample_input.yml diff --git a/docs/README.md b/docs/README.md index 0dad64f..4778fa5 100644 --- a/docs/README.md +++ b/docs/README.md @@ -2,7 +2,7 @@ ![data service logo](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/kfdrc-logo-sm.png) -LOH workflow assesses the loss of heterozygosity (LOH) in the tumor for rare germline variants. +Preprocessing LOH assesses the loss of heterozygosity (LOH) in the tumor for rare germline variants. Order of operations: This workflow runs bcftools to extract data and prepare list of locations from vcf provided and feed it into bam-readcount to compute VAF and later parse, merge germline and tumor data together. ## Inputs diff --git a/tools/run_gene_extract_list_prepare.cwl b/tools/run_gene_extract_list_prepare.cwl index ef63e57..029ca14 100644 --- a/tools/run_gene_extract_list_prepare.cwl +++ b/tools/run_gene_extract_list_prepare.cwl @@ -3,7 +3,7 @@ cwlVersion: v1.2 class: CommandLineTool id: run_gene_extract_list_prepare -label: run_germline +label: germline_tool doc: collects info from bcftool and add gene, prepare directory of list for bam-readcount tool requirements: - class: ShellCommandRequirement diff --git a/tools/run_readcount_parser.cwl b/tools/run_readcount_parser.cwl index f5fae67..b13b17a 100644 --- a/tools/run_readcount_parser.cwl +++ b/tools/run_readcount_parser.cwl @@ -3,6 +3,7 @@ cwlVersion: v1.2 class: CommandLineTool id: run_readcount_parser +label: tumor_tool doc: run bam-readcountparser the bam-readcount output and extract inner join between data from vcf (germline) and cram/bam file (tumor) requirements: - class: ShellCommandRequirement diff --git a/workflow/run_LOH_app.cwl b/workflow/run_LOH_app.cwl index b4a5a93..12f7457 100644 --- a/workflow/run_LOH_app.cwl +++ b/workflow/run_LOH_app.cwl @@ -2,22 +2,25 @@ cwlVersion: v1.2 class: Workflow -id: run_LOH_app +id: kf-loss-of-heterozygosity-preprocessing label: Kids First Loss of Heterozygosity doc: | # Kids First Loss of Heterozygosity (LOH) ![data service logo](https://github.com/d3b-center/d3b-research-workflows/raw/master/doc/kfdrc-logo-sm.png) - The Kids First Loss of Heterozygosity (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This workflow is designed to analyze LOH for family trios as well as multiple proband tumor samples. + The Kids First Loss of Heterozygosity Preprocessing (aka LOH) is a CWL workflow that assesses the loss of heterozygosity in the tumor for rare germline calls filtered by gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. This preprocessing is designed to compute variant allele frequency (VAF) for multiple proband tumor samples and can also map germline VAF for family trios if trio germline VCF file is provided. + + #### Basic info + - Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH/1.0.1 + - tested with + - Seven Bridges Cavatica Platform: https://cavatica.sbgenomics.com/ + - cwltool: https://github.com/common-workflow-language/cwltool/releases/tag/3.1.20221201130942 ### Description The Kids First Loss of Heterozygosity application is divided into two tools: Germline tool and Tumor tool. - #### Docker - Dockerfile: https://github.com/d3b-center/bixtools/tree/master/LOH - #### Germline Tool Germline tool filters germline annotations to retain variants based on gnomad_3_1_1_AF_popmax (typically < 0.01) or when gnomad_3_1_1_AF_popmax is not defined. It requires vcf file, proband sample id, ram as required inputs and peddy file as optional input which is required for family trios. It outputs variant information such as gene, chr, start, stop, ref/alt alleles, ref/alt allele depths, variant allele frequency and list of coordinates that will be an input to tumor tool. @@ -39,7 +42,7 @@ doc: | # Required participant_id: { doc: provide participant id for this run, type: string } bamscrams: { doc: tumor input file in cram or bam format with their index file, type: 'File[]' , secondaryFiles: [ { pattern: ".crai", required: false }, { pattern: ".bai", required: false } ] } - reference: { doc: human reference in fasta format with index file, type: File,secondaryFiles: [ .fai ],"sbg:suggestedValue": {class: File, path: 60639014357c3a53540ca7a3, name: Homo_sapiens_assembly38.fasta} } + reference: { doc: human reference in fasta format with index file, type: File, secondaryFiles: [ .fai ], "sbg:suggestedValue": { class: File, path: 60639014357c3a53540ca7a3, name: Homo_sapiens_assembly38.fasta, secondaryFiles: [{class: File, path: 60639016357c3a53540ca7af, name: Homo_sapiens_assembly38.fasta.fai}]} } sample_vcf_file: { doc: provide germline vcf file for this sample, type: File } # Optional minDepth: { doc: provide minDepth to consider for tumor reads, type: 'int?', default: 1 } @@ -104,10 +107,12 @@ steps: "sbg:license": Apache License 2.0 "sbg:publisher": KFDRC "sbg:categories": -- DNA +- VAF +- LOH - WGS - WXS - GVCF +- TRIOS "sbg:links": - id: 'https://github.com/d3b-center/tumor-loh-app-dev/releases/tag/v1.0.2' label: github-release \ No newline at end of file From fedd1cab99a78ed9590342f60115cc1c9a18a939 Mon Sep 17 00:00:00 2001 From: Saksham Phul Date: Thu, 29 Jun 2023 11:34:50 -0400 Subject: [PATCH 18/18] Final fix --- ..._app.cwl => kf-loss-of-heterozygosity-preprocessing-wf.cwl} | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) rename workflow/{run_LOH_app.cwl => kf-loss-of-heterozygosity-preprocessing-wf.cwl} (99%) diff --git a/workflow/run_LOH_app.cwl b/workflow/kf-loss-of-heterozygosity-preprocessing-wf.cwl similarity index 99% rename from workflow/run_LOH_app.cwl rename to workflow/kf-loss-of-heterozygosity-preprocessing-wf.cwl index 12f7457..5531c46 100644 --- a/workflow/run_LOH_app.cwl +++ b/workflow/kf-loss-of-heterozygosity-preprocessing-wf.cwl @@ -103,7 +103,8 @@ steps: minCore: minCore out: [ loh_output_file_tool,log_output ] - +$namespaces: + sbg: https://sevenbridges.com "sbg:license": Apache License 2.0 "sbg:publisher": KFDRC "sbg:categories":