Last updated: 2020-06-17
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Knit directory: apaQTL/analysis/
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Rmd | 3f63045 | brimittleman | 2019-05-24 | add code to prepare non norm qtl |
In order to compare effect sizes for the QTLs I have previously identified in an interpretable manner, I need to run the linear model with the non normalized usage. To do this I will separate the the usage (with annotation) files by chromosome and run fastqtl on these files.
library(workflowr)
This is workflowr version 1.6.0
Run ?workflowr for help getting started
library(tidyverse)
── Attaching packages ───────────────────────────────────── tidyverse 1.2.1 ──
✔ ggplot2 3.1.1 ✔ purrr 0.3.2
✔ tibble 2.1.1 ✔ dplyr 0.8.0.1
✔ tidyr 0.8.3 ✔ stringr 1.3.1
✔ readr 1.3.1 ✔ forcats 0.3.0
── Conflicts ──────────────────────────────────────── tidyverse_conflicts() ──
✖ dplyr::filter() masks stats::filter()
✖ dplyr::lag() masks stats::lag()
library(cowplot)
Attaching package: 'cowplot'
The following object is masked from 'package:ggplot2':
ggsave
countsnum= APApeak_Phenotype_GeneLocAnno.Total.5perc.CountsNumeric, APApeak_Phenotype_GeneLocAnno.Nuclear.5perc.CountsNumeric
id file= APApeak_Phenotype_GeneLocAnno.Nuclear.5perc.fc.gz, APApeak_Phenotype_GeneLocAnno.Total.5perc.fc.gz
totAnno= read.table("../data/phenotype_5perc/APApeak_Phenotype_GeneLocAnno.Total.5perc.fc.gz", stringsAsFactors = F, header = T) %>% separate(chrom, into=c("Chrchrom", "Start", "End", "ID"),sep=":") %>% mutate(Chr=str_sub(Chrchrom, 4, str_length(Chrchrom)))
colnamesTot= colnames(totAnno)[5:56]
totUsage=read.table("../data/phenotype_5perc/APApeak_Phenotype_GeneLocAnno.Total.5perc.CountsNumeric", stringsAsFactors = F, header = F, col.names = colnamesTot)
totUsageAnno=as.data.frame(cbind(Chr=totAnno$Chr, start=totAnno$Start, end=totAnno$End, ID=totAnno$ID, totUsage ))
write.table(totUsageAnno,file="../data/nonNorm_pheno/TotalUsageAllChrom.txt", col.names = T, row.names = F, quote = F, sep="\t" )
nucAnno= read.table("../data/phenotype_5perc/APApeak_Phenotype_GeneLocAnno.Nuclear.5perc.fc.gz", stringsAsFactors = F, header = T)%>% separate(chrom, into=c("Chrchrom", "Start", "End", "ID"),sep=":") %>% mutate(Chr=str_sub(Chrchrom, 4, str_length(Chrchrom)))
colnamesNuc= colnames(nucAnno)[5:56]
nucUsage=read.table("../data/phenotype_5perc/APApeak_Phenotype_GeneLocAnno.Nuclear.5perc.CountsNumeric", stringsAsFactors = F, header = F, col.names = colnamesNuc)
nucUsageAnno=as.data.frame(cbind(Chr=nucAnno$Chr, start=nucAnno$Start, end=nucAnno$End, ID=nucAnno$ID, nucUsage ))
write.table(nucUsageAnno,file="../data/nonNorm_pheno/NuclearUsageAllChrom.txt", col.names = T, row.names = F, quote = F, sep="\t" )
I will create a python script to seperate the file into each chromosome for running fastQTL.
sbatch run_sepUsagephen.sh
sbatch ZipandTabPheno.sh
sbatch ApaQTL_nominalNonnorm.sh
Concatinate files:
cat ../data/nonNorm_pheno/TotalUsageChrom*.nominal.out > ../data/nonNorm_pheno/TotalUsageChrom_Nominal_AllChrom.txt
cat ../data/nonNorm_pheno/NuclearUsageChrom*.nominal.out > ../data/nonNorm_pheno/NuclearUsageChrom_Nominal_AllChrom.txt
python qtlsPvalOppFrac.py ../data/apaQTLs/Nuclear_apaQTLs4pc_5fdr.txt ../data/nonNorm_pheno/TotalUsageChrom_Nominal_AllChrom.txt ../data/QTLoverlap_nonNorm/NuclearQTLinTotalNominal_nonNorm.txt
python qtlsPvalOppFrac.py ../data/apaQTLs/Nuclear_apaQTLs4pc_5fdr.txt ../data/nonNorm_pheno/NuclearUsageChrom_Nominal_AllChrom.txt ../data/QTLoverlap_nonNorm/NuclearQTLinNuclearNominal_nonNorm.txt
python qtlsPvalOppFrac.py ../data/apaQTLs/Total_apaQTLs4pc_5fdr.txt ../data/nonNorm_pheno/TotalUsageChrom_Nominal_AllChrom.txt ../data/QTLoverlap_nonNorm/TotalQTLinTotalNominal_nonNorm.txt
python qtlsPvalOppFrac.py ../data/apaQTLs/Total_apaQTLs4pc_5fdr.txt ../data/nonNorm_pheno/NuclearUsageChrom_Nominal_AllChrom.txt ../data/QTLoverlap_nonNorm/TotalQTLinNuclearNominal_nonNorm.txt
totAPAinNuc=read.table("../data/QTLoverlap_nonNorm/TotalQTLinNuclearNominal_nonNorm.txt", header = F, stringsAsFactors = F, col.names=c("peakID", "snp", "dist", "pval", "slope"))
nucAPAinTot=read.table("../data/QTLoverlap_nonNorm/NuclearQTLinTotalNominal_nonNorm.txt", header = F, stringsAsFactors = F, col.names=c("peakID", "snp", "dist", "pval", "slope"))
totAPAinTot=read.table("../data/QTLoverlap_nonNorm/TotalQTLinTotalNominal_nonNorm.txt", header = F, stringsAsFactors = F, col.names=c("peakID", "snp", "dist", "pval", "slope")) %>% dplyr::select(peakID, snp, slope) %>% dplyr::rename("Originalslope"=slope)
nucAPAinNuc=read.table("../data/QTLoverlap_nonNorm/NuclearQTLinNuclearNominal_nonNorm.txt", header = F, stringsAsFactors = F, col.names=c("peakID", "snp", "dist", "pval", "slope")) %>% dplyr::select(peakID, snp, slope)%>% dplyr::rename("Originalslope"=slope)
TotBoth= totAPAinNuc %>% inner_join(totAPAinTot,by=c("peakID", "snp"))
summary(lm(TotBoth$slope ~ TotBoth$Originalslope))
Call:
lm(formula = TotBoth$slope ~ TotBoth$Originalslope)
Residuals:
Min 1Q Median 3Q Max
-0.66101 -0.03117 0.00361 0.03873 0.37205
Coefficients:
Estimate Std. Error t value Pr(>|t|)
(Intercept) -0.004458 0.004247 -1.05 0.295
TotBoth$Originalslope 0.638860 0.022554 28.33 <2e-16 ***
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Residual standard error: 0.0852 on 401 degrees of freedom
Multiple R-squared: 0.6668, Adjusted R-squared: 0.6659
F-statistic: 802.4 on 1 and 401 DF, p-value: < 2.2e-16
totbothplot=ggplot(TotBoth, aes(x=Originalslope, y=slope))+geom_point() + geom_smooth(method="lm") + labs(title="Total apaQTL effect sizes", x="Effect size in Total",y="Effect size in Nucler") + geom_density_2d(col="red") + annotate("text", y=1, x=0, label="R2=.67, slope=0.64")
NucBoth= nucAPAinTot %>% inner_join(nucAPAinNuc,by=c("peakID", "snp"))
summary(lm(NucBoth$slope ~ NucBoth$Originalslope))
Call:
lm(formula = NucBoth$slope ~ NucBoth$Originalslope)
Residuals:
Min 1Q Median 3Q Max
-0.23354 -0.03332 0.00028 0.03182 0.40042
Coefficients:
Estimate Std. Error t value Pr(>|t|)
(Intercept) 0.001628 0.002910 0.559 0.576
NucBoth$Originalslope 0.775971 0.017539 44.243 <2e-16 ***
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Residual standard error: 0.06408 on 483 degrees of freedom
Multiple R-squared: 0.8021, Adjusted R-squared: 0.8017
F-statistic: 1957 on 1 and 483 DF, p-value: < 2.2e-16
Nucbothplot=ggplot(NucBoth, aes(x=Originalslope, y=slope))+geom_point() + geom_smooth(method="lm") + labs(title="Nuclear apaQTL effect sizes", x="Effect size in Nuclear",y="Effect size in Total") + geom_density_2d(col="red") + annotate("text", y=1, x=0, label="R2=.8, slope=0.78")
plot_grid(totbothplot,Nucbothplot)
For the nuclear plot I want to include the PAS we cannot test in the total fraction. To do this I will write code that only writes out the lines for the PAS not in the total fraction. I need the nominal effect sizes for the nuclear qtls in PAS we could not test in total. First I will get all of the PAS tested in total and figure out the nuclear QTLs not in this set. I will then pull those associations out of the nominal nuclear set.
python nucSpeceffectsize.py
nucSpec=read.table("../data/QTLoverlap_nonNorm/NuclearSpecQTLinNuclearNominal_nonNorm.txt",header = F, stringsAsFactors = F, col.names=c("peakID", "snp", "dist", "pval", "Originalslope")) %>% mutate(slope=0,set="Specific") %>% select(peakID, snp, dist, pval, Originalslope, slope,set)
NucBoth_set= NucBoth %>% mutate(set="Both")
NucBothwSpec=bind_rows(NucBoth_set, nucSpec)
summary(lm(NucBothwSpec$slope ~ NucBothwSpec$Originalslope))
Call:
lm(formula = NucBothwSpec$slope ~ NucBothwSpec$Originalslope)
Residuals:
Min 1Q Median 3Q Max
-0.29479 -0.03377 0.00364 0.03780 0.34739
Coefficients:
Estimate Std. Error t value Pr(>|t|)
(Intercept) -0.009236 0.002974 -3.106 0.00199 **
NucBothwSpec$Originalslope 0.664058 0.018388 36.114 < 2e-16 ***
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Residual standard error: 0.07255 on 600 degrees of freedom
Multiple R-squared: 0.6849, Adjusted R-squared: 0.6844
F-statistic: 1304 on 1 and 600 DF, p-value: < 2.2e-16
cor.test(y=NucBothwSpec$slope, x=NucBothwSpec$Originalslope)$p.value
[1] 1.336923e-152
nucspecplot=ggplot(NucBothwSpec, aes(x=Originalslope, y=slope))+geom_point(aes( col=set)) + geom_smooth(method="lm") + labs(title="Nuclear apaQTL effect sizes", x="Effect size in Nuclear",y="Effect size in Total") + annotate("text", y=1, x=0, label="R2=.68, slope=0.66",size=6) + scale_color_brewer(palette = "Dark2")+ theme(text = element_text(size=16), legend.position = "bottom")
Do this in the other fraction to see what is looks like.
python totSeceffectsize.py
totSpec=read.table("../data/QTLoverlap_nonNorm/TotalSpecQTLinTotalNominal_nonNorm.txt",header = F, stringsAsFactors = F, col.names=c("peakID", "snp", "dist", "pval", "Originalslope")) %>% mutate(slope=0,set="Specific") %>% select(peakID, snp, dist, pval, Originalslope, slope,set)
TotBoth_set= TotBoth %>% mutate(set="Both")
TotBothwSpec=bind_rows(TotBoth_set, totSpec)
summary(lm(TotBothwSpec$slope ~ TotBothwSpec$Originalslope))
Call:
lm(formula = TotBothwSpec$slope ~ TotBothwSpec$Originalslope)
Residuals:
Min 1Q Median 3Q Max
-0.64645 -0.03081 0.00337 0.03800 0.37917
Coefficients:
Estimate Std. Error t value Pr(>|t|)
(Intercept) -0.006902 0.003969 -1.739 0.0827 .
TotBothwSpec$Originalslope 0.621773 0.021776 28.553 <2e-16 ***
---
Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1
Residual standard error: 0.08339 on 441 degrees of freedom
Multiple R-squared: 0.649, Adjusted R-squared: 0.6482
F-statistic: 815.2 on 1 and 441 DF, p-value: < 2.2e-16
totspecplot=ggplot(TotBothwSpec, aes(x=Originalslope, y=slope))+geom_point(aes( col=set)) + geom_smooth(method="lm") + labs(title="Total apaQTL effect sizes", x="Effect size in Total",y="Effect size in Nuclear") + annotate("text", y=1, x=0, label="R2=.65, slope=0.62", size=6) + scale_color_brewer(palette = "Dark2") + theme(text = element_text(size=16), legend.position = "bottom")
totspecplot
grideffectplot=plot_grid(nucspecplot,totspecplot)
grideffectplot
NucBothwSpec_onlyboth=NucBothwSpec %>% filter(set=="Both")
nucspecplot2=ggplot(NucBothwSpec_onlyboth, aes(x=Originalslope, y=slope))+geom_point(col="#1B9E77") + geom_smooth(method="lm") + labs( x="Effect size in Nuclear",y="Effect size in Total") + theme(text = element_text(size=16), legend.position = "bottom") + geom_abline(slope = 1)
nucspecplot2
TotBothwSpec %>% filter(set=="Specific") %>% nrow()
[1] 40
NucBothwSpec %>% filter(set=="Specific") %>% nrow()
[1] 117
prop.test(x=c(117,40), n=c(565,378), alternative ="greater")
2-sample test for equality of proportions with continuity
correction
data: c(117, 40) out of c(565, 378)
X-squared = 16.012, df = 1, p-value = 3.147e-05
alternative hypothesis: greater
95 percent confidence interval:
0.06079568 1.00000000
sample estimates:
prop 1 prop 2
0.2070796 0.1058201
specific PAS:
totalPAS=read.table("../data/PAS/APApeak_Peaks_GeneLocAnno.Total.5perc.sort.bed",stringsAsFactors = F,col.names = c("chr", "start","end","name", "score", "strand"))
nuclearPAS=read.table("../data/PAS/APApeak_Peaks_GeneLocAnno.Nuclear.5perc.sort.bed",stringsAsFactors = F,col.names = c("chr", "start","end","name", "score", "strand"))
totalSpec=totalPAS %>% anti_join(nuclearPAS,by="name")
40/nrow(totalSpec)
[1] 0.01509434
sessionInfo()
R version 3.5.1 (2018-07-02)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Scientific Linux 7.4 (Nitrogen)
Matrix products: default
BLAS/LAPACK: /software/openblas-0.2.19-el7-x86_64/lib/libopenblas_haswellp-r0.2.19.so
locale:
[1] LC_CTYPE=en_US.UTF-8 LC_NUMERIC=C
[3] LC_TIME=en_US.UTF-8 LC_COLLATE=en_US.UTF-8
[5] LC_MONETARY=en_US.UTF-8 LC_MESSAGES=en_US.UTF-8
[7] LC_PAPER=en_US.UTF-8 LC_NAME=C
[9] LC_ADDRESS=C LC_TELEPHONE=C
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C
attached base packages:
[1] stats graphics grDevices utils datasets methods base
other attached packages:
[1] cowplot_0.9.4 forcats_0.3.0 stringr_1.3.1 dplyr_0.8.0.1
[5] purrr_0.3.2 readr_1.3.1 tidyr_0.8.3 tibble_2.1.1
[9] ggplot2_3.1.1 tidyverse_1.2.1 workflowr_1.6.0
loaded via a namespace (and not attached):
[1] tidyselect_0.2.5 haven_1.1.2 lattice_0.20-38
[4] colorspace_1.3-2 generics_0.0.2 htmltools_0.3.6
[7] yaml_2.2.0 rlang_0.4.0 later_0.7.5
[10] pillar_1.3.1 glue_1.3.0 withr_2.1.2
[13] RColorBrewer_1.1-2 modelr_0.1.2 readxl_1.1.0
[16] plyr_1.8.4 munsell_0.5.0 gtable_0.2.0
[19] cellranger_1.1.0 rvest_0.3.2 evaluate_0.12
[22] labeling_0.3 knitr_1.20 httpuv_1.4.5
[25] broom_0.5.1 Rcpp_1.0.4.6 promises_1.0.1
[28] scales_1.0.0 backports_1.1.2 jsonlite_1.6
[31] fs_1.3.1 hms_0.4.2 digest_0.6.18
[34] stringi_1.2.4 grid_3.5.1 rprojroot_1.3-2
[37] cli_1.1.0 tools_3.5.1 magrittr_1.5
[40] lazyeval_0.2.1 crayon_1.3.4 whisker_0.3-2
[43] pkgconfig_2.0.2 MASS_7.3-51.1 xml2_1.2.0
[46] lubridate_1.7.4 assertthat_0.2.0 rmarkdown_1.10
[49] httr_1.3.1 rstudioapi_0.10 R6_2.3.0
[52] nlme_3.1-137 git2r_0.26.1 compiler_3.5.1