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Rscript ../scripts/ssgsea_enrich_diff.r -h usage: ../scripts/ssgsea_enrich_diff.r [-h] -g GMTFILE -i EXPR -m META [-n GROUP_NAME] [--log2] [-t method] [-k kcdf] [--group1 GROUP1] [--group2 GROUP2] [-p PVALUECUTOFF] [--no_diff] [-o OUTDIR] [-f PREFIX] Gene set variation analysis (GSVA):https://www.亿速云.com/article/1586 optional arguments: -h, --help show this help message and exit -g GMTFILE, --gmtfile GMTFILE GSEA gmtfile function class file[required] -i EXPR, --expr EXPR Input gene expression file path[required] -m META, --meta META Input the clinical information file path that contains the grouping[required] -n GROUP_NAME, --group_name GROUP_NAME Specifies the column name that contains grouping information[optional,default:m6acluster] --log2 Whether to perform log2 processing[optional,default:False] -t method, --method method Method to employ in the estimation of gene-set enrichment scores per sample. By default this is set to gsva (Hänzelmann et al, 2013) and other options are ssgsea (Barbie et al, 2009), zscore (Lee et al, 2008) or plage (Tomfohr et al, 2005). The latter two standardize first expression profiles into z-scores over the samples and, in the case of zscore, it combines them together as their sum divided by the square-root of the size of the gene set, while in the case of plage they are used to calculate the singular value decomposition (SVD) over the genes in the gene set and use the coefficients of the first right- singular vector as pathway activity profile[default gsva] -k kcdf, --kcdf kcdf Character string denoting the kernel to use during the non-parametric estimation of the cumulative distribution function of expression levels across samples when method="gsva". By default, kcdf="Gaussian" which is suitable when input expression values are continuous, such as microarray fluorescent units in logarithmic scale, RNA-seq log- CPMs, log-RPKMs or log-TPMs. When input expression values are integer counts, such as those derived from RNA-seq experiments, then this argument should be set to kcdf="Poisson"[default Gaussian] --group1 GROUP1 Designate the first group[optional,default C1] --group2 GROUP2 Designate the second group[optional,default C2] -p PVALUECUTOFF, --pvalueCutoff PVALUECUTOFF pvalue cutoff on enrichment tests to report[optional,default:0.05] --no_diff No screening was performed based on the difference analysis results[optional,default:False] -o OUTDIR, --outdir OUTDIR output file directory[optional,default cwd] -f PREFIX, --prefix PREFIX out file name prefix[optional,default kegg]
-g 参考基因集,如从MSigDB下载的KEGG基因集c2.cp.kegg.v6.2.symbols.gmt
KEGG_GLYCOLYSIS_GLUCONEOGENESIS | http://www.broadinstitute.org/gsea/msigdb/cards/KEGG_GLYCOLYSIS_GLUCONEOGENESIS | ACSS2 | GCK |
KEGG_CITRATE_CYCLE_TCA_CYCLE | http://www.broadinstitute.org/gsea/msigdb/cards/KEGG_CITRATE_CYCLE_TCA_CYCLE | IDH3B | DLST |
KEGG_PENTOSE_PHOSPHATE_PATHWAY | http://www.broadinstitute.org/gsea/msigdb/cards/KEGG_PENTOSE_PHOSPHATE_PATHWAY | RPE | RPIA |
-i 基因表达矩阵
ID | TCGA-A3-3319-01A-02R-1325-07 | TCGA-A3-3323-01A-02R-1325-07 |
YTHDC2 | 16.5128725081007 | 20.6535652352011 |
ELAVL1 | 44.3876796198438 | 31.8729000784291 |
-m 包含样本分组信息的样本信息文件
barcode | patient | sample |
TCGA-BP-4766-01A-01R-1289-07 | TCGA-BP-4766 | TCGA-BP-4766-01A |
TCGA-A3-3352-01A-01R-0864-07 | TCGA-A3-3352 | TCGA-A3-3352-01A |
--log2 是否对基因表达矩阵进行log2转换
-n 指定样本信息文件中分组信息的列名
--group1 --group2 指定分组组名
-t 指定用于估计基因集的方法,默认为gsva
-k 指定gsva函数中的kcdf参数,使用read count数据时一般设为“Poisson”,使用log后的TPM等数据时一般就用默认值“Gaussian”
-p 指定p的阈值
--no_diff GSVA会先将表达矩阵转换成富集分数矩阵,然后再通过差异表达分析筛选富集结果,设置这个参数则不对富集结果进行筛选
Rscript ../scripts/ssgsea_enrich_diff.r -g enrich/c2.cp.kegg.v6.2.symbols.gmt \ -i ../02.sample_select/TCGA-KIRC_gene_expression_TPM_immu.tsv -m metadata_group.tsv \ -n m6acluster --log2 --group1 C1 --group2 C2 -p 0.05 -o enrich/C1_vs_C2
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