GTEx_genes
ERM
2023-06-14
Last updated: 2023-06-14
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Knit directory: Cardiotoxicity/
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| Rmd | ff02989 | reneeisnowhere | 2023-06-14 | update for website |
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| Rmd | e9f1a70 | reneeisnowhere | 2023-06-14 | update for website |
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GTEx egenes and analysis
library(ComplexHeatmap)
library(tidyverse)
library(ggsignif)
library(biomaRt)
library(RColorBrewer)
library(cowplot)
library(scales)
library(sjmisc)
library(kableExtra)
library(broom)
library(ggstats)Data import
toplistall<- read.csv("output/toplistall.csv", row.names = 1)
my_exp_genes <- read.csv("data/backGL.txt")
egenes_set <- read.csv("output/egenes_set.csv",row.names = 1)
egenes_hgnc <- read.csv("output/egenes_hgnc.csv",row.names = 1)
GTEx_genes <- read.csv("data/GTEx_gene_list.csv",row.names = 1)
not_eqtls <- read.csv("output/not_eqtls_GTEX.csv",row.names = 1)
heart_gtex <- read.csv("output/heart_gtex.csv",row.names = 1)
egenes <- read.csv("output/egenes.csv",row.names = 1)Obtaining the GTEx data set
I downloaded the GTEx_Analysis_v8.metasoft.txt.gz files from the Consortium at https://www.gtexportal.org/home/datasets .
I the extracted the Heart_Left_Ventricle.v8.egenes.txt file and uploaded into R under the data folder.
heart_gtex <-
readr::read_delim("data/Heart_Left_Ventricle.v8.egenes.txt",
delim = "\t",
escape_double = FALSE,
trim_ws = TRUE)
egenes <- heart_gtex %>%
dplyr::select(gene_id, gene_name, qval) %>%
filter(qval<0.05) %>%
separate(gene_id, into =c('ensembl_gene_id', 'gene_version'))
not_eqtl <- heart_gtex %>%
dplyr::select(gene_id, gene_name, qval) %>%
filter(qval>0.05) %>%
separate(gene_id, into =c('ensembl_gene_id', 'gene_version'))
egenes_set <- getBM(attributes=my_attributes,
filters ='ensembl_gene_id',
values =egenes$ensembl_gene_id,
mart = ensembl)
egenes_hgnc <- getBM(attributes=my_attributes,
filters ='hgnc_symbol',
values =egenes$gene_name,
mart = ensembl)
not_eqtl_set <- getBM(attributes=my_attributes,
filters ='ensembl_gene_id',
values =not_eqtl$ensembl_gene_id,
mart = ensembl)
not_eqtls <- not_eqtl_set %>%
distinct(entrezgene_id,.keep_all = TRUE) %>%
filter(entrezgene_id %in% my_exp_genes$ENTREZID)
##6711 not_eqtls
GTEx_genes <- egenes_set %>%
distinct(entrezgene_id,.keep_all = TRUE)This file contains several columns gene_id, gene_name, gene_chr, gene_start, gene_end, strand, num_var, beta_shape1, beta_shape2, true_df, pval_true_df, variant_id, tss_distance, chr, variant_pos, ref, alt, num_alt_per_site, rs_id_dbSNP151_GRCh38p7, minor_allele_samples, minor_allele_count, maf, ref_factor, pval_nominal, slope, slope_se, pval_perm, pval_beta, qval, pval_nominal_threshold, log2_aFC, log2_aFC_lower, log2_aFC_upper.
I then chose the the ‘gene_id’,‘gene_name’, and ‘qval’ columns. This
left me with 21353 genes. Next I filtered the tissue specific expressed
genes using a the ‘qval < 0.05’ for a total of 9642. I then took the
gene_name column and used biomart to convert to ‘entrezgene_id’.
Because results vary by which way I look up genes in BioMart, I tested
both egenes using ensemble_gene_id and hgnc_symbol columns. I found 7813
for the ensemble_gene_ set and 7271 for the hgnc_symbol set.
I went with using ensemble_gene_id because I found ~600 more genes
overall than using the hgnc_symbol filter.
GTEx <- intersect(GTEx_genes$entrezgene_id,my_exp_genes$ENTREZID)
nQTL <- not_eqtls
# '%!in%' <- function(x,y)!('%in%'(x,y))
# nQTL <- toplistall %>%
# filter(adj.P.Val<0.05) %>%
# distinct(ENTREZID) %>%
# dplyr:: filter(!ENTREZID %in%GTEx)
# testset <- toplistall %>%
# filter(adj.P.Val<0.05) %>%
# select(ENTREZID) %>% distinct(ENTREZID) %>%
# dplyr:: filter(ENTREZID %in%GTEx_genes$entrezgene_id)To find out how many genes from the gtex egenes were expressed in my data, I intersected my expressed genes list of 14084 genes with the GTEx_genes and found 6261 genes were shared between them. I called this set ‘GTEx’. Using the other eGenes from GTEx, I made another set intersected with my expressed gene set called ‘nQTL’. This nQTL set contains 6711 genes. Next, I then took my DEG top list and filtered out genes with an adj.P.value < 0.05.
drug_palspc <- c("#8B006D","#DF707E","#8B006D","#DF707E")
GTEX_table <- toplistall %>%
mutate(id = as.factor(id)) %>%
mutate(time=factor(time, levels=c("3_hours","24_hours"))) %>%
dplyr::filter(adj.P.Val <0.05) %>%
mutate(nQTL=if_else(ENTREZID %in% nQTL$entrezgene_id,'nQTL_y','nQTL_no')) %>%
mutate(GTEx=if_else(ENTREZID %in%GTEx,"GTEx_y","GTEx_no")) %>%
dplyr::select( id, time,ENTREZID,GTEx,nQTL) %>%
group_by(id,time,GTEx,nQTL) %>%
tally() %>% as.data.frame()#
nQTL_table <- toplistall %>%
mutate(id = as.factor(id)) %>%
mutate(time=factor(time, levels=c("3_hours","24_hours"))) %>%
dplyr::filter(adj.P.Val <0.05) %>%
mutate(nQTL=if_else(ENTREZID %in%nQTL$entrezgene_id,'nQTL_y','nQTL_no')) %>%
dplyr::select( id, time,ENTREZID,nQTL) %>%
group_by(id,time,nQTL) %>%
summarize(total=n()) %>% as.data.frame()
GTEX_table_chi <- toplistall %>%
mutate(id = as.factor(id)) %>%
mutate(time=factor(time, levels=c("3_hours","24_hours"))) %>%
dplyr::filter(adj.P.Val <0.05) %>%
mutate(nQTL=if_else(ENTREZID %in%nQTL$entrezgene_id, 'nQTL_y','nQTL_no')) %>%
mutate(GTEx=if_else(ENTREZID %in%GTEx,"GTEx_y","GTEx_no")) %>%
dplyr::select( id, time,ENTREZID,GTEx,nQTL) %>%
group_by(id,time) %>%
summarise(pvalue= chisq.test(nQTL, GTEx)$p.value)
toplistall %>%
mutate(id = as.factor(id)) %>%
mutate(time=factor(time, levels=c("3_hours","24_hours"))) %>%
dplyr::filter(adj.P.Val <0.05) %>%
mutate(nQTL= if_else(ENTREZID %in% nQTL$entrezgene_id,'nQTL_y','nQTL_no')) %>%
mutate(GTEx=if_else(ENTREZID %in%GTEx,"GTEx_y","GTEx_no")) %>%
dplyr::select( id, time,ENTREZID,GTEx,nQTL) %>%
group_by(id,time,nQTL,GTEx) %>%
# summarize(value=n())#%>%
pivot_longer(!c(id,time, ENTREZID)) %>%
ggplot(., aes(x=name))+
geom_bar(aes(fill=value),position="fill", stat="count")+
# guides(fill="none")+
facet_wrap(time~id,nrow=2,ncol=4)+
theme_classic()+
# scale_color_manual(values=drug_palNoVeh[c(1,2)])+
scale_fill_manual(values=drug_palspc)+
scale_y_continuous(label = scales::percent)
GTEX_table %>% kable(., caption= "Genes from Gtex that overlap and do not overlap DE genes, and all not_gtex genes that overlap and do not overlap DE genes")%>%
kable_paper("striped", full_width = FALSE) %>%
kable_styling(full_width = FALSE, position = "left",bootstrap_options = c("striped"),font_size = 18) %>%
scroll_box(width = "60%", height = "800px")| id | time | GTEx | nQTL | n |
|---|---|---|---|---|
| Daunorubicin | 3_hours | GTEx_no | nQTL_no | 22 |
| Daunorubicin | 3_hours | GTEx_no | nQTL_y | 343 |
| Daunorubicin | 3_hours | GTEx_y | nQTL_no | 190 |
| Daunorubicin | 24_hours | GTEx_no | nQTL_no | 511 |
| Daunorubicin | 24_hours | GTEx_no | nQTL_y | 3321 |
| Daunorubicin | 24_hours | GTEx_y | nQTL_no | 3028 |
| Daunorubicin | 24_hours | GTEx_y | nQTL_y | 4 |
| Doxorubicin | 3_hours | GTEx_no | nQTL_y | 14 |
| Doxorubicin | 3_hours | GTEx_y | nQTL_no | 2 |
| Doxorubicin | 24_hours | GTEx_no | nQTL_no | 471 |
| Doxorubicin | 24_hours | GTEx_no | nQTL_y | 3168 |
| Doxorubicin | 24_hours | GTEx_y | nQTL_no | 2875 |
| Doxorubicin | 24_hours | GTEx_y | nQTL_y | 2 |
| Epirubicin | 3_hours | GTEx_no | nQTL_no | 6 |
| Epirubicin | 3_hours | GTEx_no | nQTL_y | 137 |
| Epirubicin | 3_hours | GTEx_y | nQTL_no | 77 |
| Epirubicin | 24_hours | GTEx_no | nQTL_no | 458 |
| Epirubicin | 24_hours | GTEx_no | nQTL_y | 3058 |
| Epirubicin | 24_hours | GTEx_y | nQTL_no | 2684 |
| Epirubicin | 24_hours | GTEx_y | nQTL_y | 2 |
| Mitoxantrone | 3_hours | GTEx_no | nQTL_no | 3 |
| Mitoxantrone | 3_hours | GTEx_no | nQTL_y | 42 |
| Mitoxantrone | 3_hours | GTEx_y | nQTL_no | 13 |
| Mitoxantrone | 24_hours | GTEx_no | nQTL_no | 104 |
| Mitoxantrone | 24_hours | GTEx_no | nQTL_y | 689 |
| Mitoxantrone | 24_hours | GTEx_y | nQTL_no | 534 |
GTEX_table_chi %>%
kable(., caption= "Genes from Gtex that overlap and do not overlap DE genes, and all not_gtex genes that overlap and do not overlap DE genes")%>%
kable_paper("striped", full_width = FALSE) %>%
kable_styling(full_width = FALSE, position = "left",bootstrap_options = c("striped"),font_size = 18) %>%
scroll_box(width = "60%", height = "800px")| id | time | pvalue |
|---|---|---|
| Daunorubicin | 3_hours | 0.0000000 |
| Daunorubicin | 24_hours | 0.0000000 |
| Doxorubicin | 3_hours | 0.0042747 |
| Doxorubicin | 24_hours | 0.0000000 |
| Epirubicin | 3_hours | 0.0000000 |
| Epirubicin | 24_hours | 0.0000000 |
| Mitoxantrone | 3_hours | 0.0000000 |
| Mitoxantrone | 24_hours | 0.0000000 |
The next step is to wrangle the data so that I can test the difference between the proportions of significantly DE genes found in the GTEx and nQTLs.
sessionInfo()R version 4.2.2 (2022-10-31 ucrt)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 10 x64 (build 19045)
Matrix products: default
locale:
[1] LC_COLLATE=English_United States.utf8
[2] LC_CTYPE=English_United States.utf8
[3] LC_MONETARY=English_United States.utf8
[4] LC_NUMERIC=C
[5] LC_TIME=English_United States.utf8
attached base packages:
[1] grid stats graphics grDevices utils datasets methods
[8] base
other attached packages:
[1] ggstats_0.3.0 broom_1.0.5 kableExtra_1.3.4
[4] sjmisc_2.8.9 scales_1.2.1 cowplot_1.1.1
[7] RColorBrewer_1.1-3 biomaRt_2.52.0 ggsignif_0.6.4
[10] lubridate_1.9.2 forcats_1.0.0 stringr_1.5.0
[13] dplyr_1.1.2 purrr_1.0.1 readr_2.1.4
[16] tidyr_1.3.0 tibble_3.2.1 ggplot2_3.4.2
[19] tidyverse_2.0.0 ComplexHeatmap_2.12.1 workflowr_1.7.0
loaded via a namespace (and not attached):
[1] colorspace_2.1-0 rjson_0.2.21 sjlabelled_1.2.0
[4] rprojroot_2.0.3 circlize_0.4.15 XVector_0.36.0
[7] GlobalOptions_0.1.2 fs_1.6.2 clue_0.3-64
[10] rstudioapi_0.14 farver_2.1.1 bit64_4.0.5
[13] AnnotationDbi_1.58.0 fansi_1.0.4 xml2_1.3.4
[16] codetools_0.2-19 doParallel_1.0.17 cachem_1.0.8
[19] knitr_1.43 jsonlite_1.8.5 cluster_2.1.4
[22] dbplyr_2.3.2 png_0.1-8 compiler_4.2.2
[25] httr_1.4.6 backports_1.4.1 fastmap_1.1.1
[28] cli_3.6.1 later_1.3.1 htmltools_0.5.5
[31] prettyunits_1.1.1 tools_4.2.2 gtable_0.3.3
[34] glue_1.6.2 GenomeInfoDbData_1.2.8 rappdirs_0.3.3
[37] Rcpp_1.0.10 Biobase_2.56.0 jquerylib_0.1.4
[40] vctrs_0.6.2 Biostrings_2.64.1 svglite_2.1.1
[43] iterators_1.0.14 insight_0.19.2 xfun_0.39
[46] ps_1.7.5 rvest_1.0.3 timechange_0.2.0
[49] lifecycle_1.0.3 XML_3.99-0.14 getPass_0.2-2
[52] zlibbioc_1.42.0 hms_1.1.3 promises_1.2.0.1
[55] parallel_4.2.2 yaml_2.3.7 curl_5.0.1
[58] memoise_2.0.1 sass_0.4.6 stringi_1.7.12
[61] RSQLite_2.3.1 highr_0.10 S4Vectors_0.34.0
[64] foreach_1.5.2 BiocGenerics_0.42.0 filelock_1.0.2
[67] shape_1.4.6 GenomeInfoDb_1.32.4 rlang_1.1.1
[70] pkgconfig_2.0.3 systemfonts_1.0.4 matrixStats_1.0.0
[73] bitops_1.0-7 evaluate_0.21 labeling_0.4.2
[76] bit_4.0.5 processx_3.8.1 tidyselect_1.2.0
[79] magrittr_2.0.3 R6_2.5.1 IRanges_2.30.1
[82] generics_0.1.3 DBI_1.1.3 pillar_1.9.0
[85] whisker_0.4.1 withr_2.5.0 KEGGREST_1.36.3
[88] RCurl_1.98-1.12 crayon_1.5.2 utf8_1.2.3
[91] BiocFileCache_2.4.0 tzdb_0.4.0 rmarkdown_2.22
[94] GetoptLong_1.0.5 progress_1.2.2 blob_1.2.4
[97] callr_3.7.3 git2r_0.32.0 digest_0.6.31
[100] webshot_0.5.4 httpuv_1.6.11 stats4_4.2.2
[103] munsell_0.5.0 viridisLite_0.4.2 bslib_0.5.0