QTL Analysis - binary [ICI-Early vs PBS & ICI-Late ]
Belinda Cornes
2022-04-15
Last updated: 2022-04-15
Checks: 5 2
Knit directory: Serreze-T1D_Workflow/
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Unstaged changes:
Modified: analysis/4.1.1_qtl.analysis_binary_ici-early.vs.pbs_5.batches.Rmd
Modified: analysis/4.1.1_qtl.analysis_binary_ici-early.vs.pbs_5.batches_mis.Rmd
Modified: analysis/4.1.1_qtl.analysis_binary_ici-early.vs.pbs_snpsqc_5.batches.Rmd
Modified: analysis/4.1.1_qtl.analysis_binary_ici-early.vs.pbs_snpsqc_5.batches_mis.Rmd
Modified: analysis/4.1.1_qtl.analysis_binary_ici-early.vs.pbs_snpsqc_dis_no-x_updated_5.batches.Rmd
Modified: analysis/4.1.1_qtl.analysis_binary_ici-early.vs.pbs_snpsqc_dis_no-x_updated_5.batches_mis.Rmd
Modified: analysis/4.1.1_qtl.analysis_binary_ici.vs.eoi_snpsqc_dis_no-x_updated.Rmd
Modified: analysis/4.1.1_qtl.analysis_binary_ici.vs.pbs_snpsqc_dis_no-x_updated.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici-early.vs.pbs_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici-early.vs.pbs_pheno.corrected.cleaned_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici-early.vs.pbs_snpsqc_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici-early.vs.pbs_snpsqc_pheno.corrected.cleaned_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici-early.vs.pbs_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici-early.vs.pbs_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.eoi_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.eoi_pheno.corrected.cleaned_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.eoi_snpsqc_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.eoi_snpsqc_pheno.corrected.cleaned_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.eoi_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.eoi_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.pbs_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.pbs_pheno.corrected.cleaned_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.pbs_snpsqc_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.pbs_snpsqc_pheno.corrected.cleaned_5.batches_mis.Rmd
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Modified: analysis/4.1.2_qtl.analysis_cont_age_ici.vs.pbs_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici-early.vs.pbs_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici-early.vs.pbs_pheno.corrected.cleaned_5.batches_mis.Rmd
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Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici-early.vs.pbs_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici-early.vs.pbs_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.eoi_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.eoi_pheno.corrected.cleaned_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.eoi_snpsqc_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.eoi_snpsqc_pheno.corrected.cleaned_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.eoi_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.eoi_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.pbs_pheno.corrected.cleaned_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.pbs_pheno.corrected.cleaned_5.batches_mis.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.pbs_snpsqc_pheno.corrected.cleaned_5.batches.Rmd
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Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.pbs_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches.Rmd
Modified: analysis/4.1.2_qtl.analysis_cont_rz.age_ici.vs.pbs_snpsqc_pheno.corrected.cleaned_dis_no-xk_5.batches_mis.Rmd
Modified: analysis/index_5.batches.Rmd
Modified: analysis/index_5.batches_additional.Rmd
Note that any generated files, e.g. HTML, png, CSS, etc., are not included in this status report because it is ok for generated content to have uncommitted changes.
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Loading Data
We will load the data and subset indivials out that are in the groups of interest. We will create a binary phenotype from this (PBS & ICI-Late ==0, ICI-Early == 1).
load("data/gm_allqc_5.batches.RData")
#gm_allqc
gm=gm_allqc
gmObject of class cross2 (crosstype "bc")
Total individuals 308
No. genotyped individuals 308
No. phenotyped individuals 308
No. with both geno & pheno 308
No. phenotypes 1
No. covariates 6
No. phenotype covariates 0
No. chromosomes 20
Total markers 34537
No. markers by chr:
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
2643 2629 1857 1890 1774 1941 1672 1627 1878 1176 1871 1300 1549 1578 1257 935
17 18 19 X
501 913 1014 4532 table(gm$covar$"diabetic status")
Both Projects Diabetes Proj ICI-No Diabetes Myocarditis Spont. T1D
1 3 164 2 34
T1D <17 T1D<17 T1D>17
36 51 17 table(gm$covar$group)
(A2 Parental) (DQ8 Parental) B6.g7 Parental EOI F1
2 2 1 164 1
ICI PBS
104 34 #ICI-Early vs (PBS-T1D + ICI > 17 weeks of age)
##extracting animals with ici-early and pbs group status
miceinfo <- gm$covar[gm$covar$group == "PBS" | gm$covar$group == "ICI",]
table(miceinfo$group)
ICI PBS
104 34 mice.ids <- rownames(miceinfo)
gm <- gm[mice.ids]
gmObject of class cross2 (crosstype "bc")
Total individuals 138
No. genotyped individuals 138
No. phenotyped individuals 138
No. with both geno & pheno 138
No. phenotypes 1
No. covariates 6
No. phenotype covariates 0
No. chromosomes 20
Total markers 34537
No. markers by chr:
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
2643 2629 1857 1890 1774 1941 1672 1627 1878 1176 1871 1300 1549 1578 1257 935
17 18 19 X
501 913 1014 4532 table(gm$covar$"diabetic status")
Spont. T1D T1D <17 T1D<17 T1D>17
34 36 51 17 table(gm$covar$group)
ICI PBS
104 34 #Creating a new lable for pbs+ici late and ici-early
table(gm$covar$group)
ICI PBS
104 34 gm$covar$group.new <- ifelse(gm$covar$"diabetic status" == "T1D>17" | gm$covar$group == "PBS", "PBS_ICI.Late", "ICI.Early")
#binary Yes/No T1D
gm$covar$ICI.Early.vs.PBS_ICI.Late <- ifelse(gm$covar$group.new == "PBS_ICI.Late", 0, 1)
table(gm$covar$"diabetic status")
Spont. T1D T1D <17 T1D<17 T1D>17
34 36 51 17 table(gm$covar$group)
ICI PBS
104 34 table(gm$covar$group.new)
ICI.Early PBS_ICI.Late
87 51 table(gm$covar$ICI.Early.vs.PBS_ICI.Late)
0 1
51 87 gm.full <- gm
##removing problmetic marker
gm <- drop_markers(gm, "UNCHS013106")
##dropping monomorphic markers within the dataset
g <- do.call("cbind", gm$geno)
gf_mar <- t(apply(g, 2, function(a) table(factor(a, 1:2))/sum(a != 0)))
#gn_mar <- t(apply(g, 2, function(a) table(factor(a, 1:2))))
gf_mar <- gf_mar[gf_mar[,2] != "NaN",]
count <- rowSums(gf_mar <=0.05)
low_freq_df <- merge(as.data.frame(gf_mar),as.data.frame(count), by="row.names",all=T)
low_freq_df[is.na(low_freq_df)] <- ''
low_freq_df <- low_freq_df[low_freq_df$count == 1,]
rownames(low_freq_df) <- low_freq_df$Row.names
low_freq <- find_markerpos(gm, rownames(low_freq_df))
low_freq$id <- rownames(low_freq)
nrow(low_freq)[1] 7989low_freq_bad <- merge(low_freq,low_freq_df, by="row.names",all=T)
names(low_freq_bad)[1] <- c("marker")
gf_mar <- gf_mar[gf_mar[,2] != "NaN",]
MAF <- apply(gf_mar, 1, function(x) min(x))
MAF <- as.data.frame(MAF)
MAF$index <- 1:nrow(gf_mar)
gf_mar_maf <- merge(gf_mar,as.data.frame(MAF), by="row.names")
gf_mar_maf <- gf_mar_maf[order(gf_mar_maf$index),]
gfmar <- NULL
gfmar$gfmar_mar_0 <- sum(gf_mar_maf$MAF==0)
gfmar$gfmar_mar_1 <- sum(gf_mar_maf$MAF< 0.01)
gfmar$gfmar_mar_5 <- sum(gf_mar_maf$MAF< 0.05)
gfmar$gfmar_mar_10 <- sum(gf_mar_maf$MAF< 0.10)
gfmar$gfmar_mar_15 <- sum(gf_mar_maf$MAF< 0.15)
gfmar$gfmar_mar_25 <- sum(gf_mar_maf$MAF< 0.25)
gfmar$gfmar_mar_50 <- sum(gf_mar_maf$MAF< 0.50)
gfmar$total_snps <- nrow(as.data.frame(gf_mar_maf))
gfmar <- t(as.data.frame(gfmar))
gfmar <- as.data.frame(gfmar)
gfmar$count <- gfmar$V1
gfmar[c(2)] %>%
kable(escape = F,align = c("ccccccccc"),linesep ="\\hline") %>%
kable_styling(full_width = F) %>%
kable_styling("striped", full_width = F) %>%
row_spec(8 ,bold=T,color= "white",background = "black")| count | |
|---|---|
| gfmar_mar_0 | 3814 |
| gfmar_mar_1 | 4068 |
| gfmar_mar_5 | 7989 |
| gfmar_mar_10 | 8527 |
| gfmar_mar_15 | 8598 |
| gfmar_mar_25 | 9313 |
| gfmar_mar_50 | 33753 |
| total_snps | 34537 |
gm_qc <- drop_markers(gm, low_freq_bad$marker)
gm_qc <- drop_nullmarkers(gm_qc)
gm = gm_qc
gmObject of class cross2 (crosstype "bc")
Total individuals 138
No. genotyped individuals 138
No. phenotyped individuals 138
No. with both geno & pheno 138
No. phenotypes 1
No. covariates 8
No. phenotype covariates 0
No. chromosomes 20
Total markers 26548
No. markers by chr:
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
2374 2374 1623 1691 1568 1703 1464 1468 1685 974 1671 1134 1377 1411 876 751
17 18 19 X
330 770 920 384 markers <- marker_names(gm)
gmapdf <- read.csv("/Users/corneb/Documents/MyJax/CS/Projects/Serreze/haplotype.reconstruction/output_5.batches/genetic_map.csv")
pmapdf <- read.csv("/Users/corneb/Documents/MyJax/CS/Projects/Serreze/haplotype.reconstruction/output_5.batches/physical_map.csv")
#mapdf <- merge(gmapdf,pmapdf, by=c("marker","chr"), all=T)
#rownames(mapdf) <- mapdf$marker
#mapdf <- mapdf[markers,]
#names(mapdf) <- c('marker','chr','gmapdf','pmapdf')
#mapdfnd <- mapdf[!duplicated(mapdf[c(2:3)]),]
pr.qc <- calc_genoprob(gm)Genome-wide scan
For each of the phenotype analyzed, permutations were used for each model to obtain genome-wide LOD significance threshold for p < 0.01, p < 0.05, p < 0.10, respectively, separately for X and automsomes (A).
The table shows the estimated significance thresholds from permutation test.
We also looked at the kinship to see how correlated each sample is. Kinship values between pairs of samples range between 0 (no relationship) and 1.0 (completely identical). The darker the colour the more indentical the pairs are.
Xcovar <- get_x_covar(gm)
#addcovar = model.matrix(~Sex, data = covars)[,-1]
#K <- calc_kinship(pr.qc, type = "loco")
#heatmap(K[[1]])
#K.overall <- calc_kinship(pr.qc, type = "overall")
#heatmap(K.overall)
kinship <- calc_kinship(pr.qc)
heatmap(kinship)
#operm <- scan1perm(pr.qc, gm$covar$phenos, Xcovar=Xcovar, n_perm=2000)
#operm <- scan1perm(pr.qc, gm$covar$phenos, addcovar = addcovar, n_perm=2000)
#operm <- scan1perm(pr.qc, gm$covar$phenos, n_perm=2000)
operm <- scan1perm(pr.qc, gm$covar["ICI.Early.vs.PBS_ICI.Late"], model="binary", n_perm=10, perm_Xsp=TRUE, chr_lengths=chr_lengths(gm$gmap))
summary_table<-data.frame(unclass(summary(operm, alpha=c(0.01, 0.05, 0.1))))
names(summary_table) <- c("autosomes","X")
summary_table$significance.level <- rownames(summary_table)
rownames(summary_table) <- NULL
summary_table[c(3,1:2)] %>%
kable(escape = F,align = c("ccc")) %>%
kable_styling("striped", full_width = T) %>%
column_spec(1, bold=TRUE)| significance.level | autosomes | X |
|---|---|---|
| 0.01 | 3.519203 | 3.401153 |
| 0.05 | 3.305096 | 3.237562 |
| 0.1 | 3.036809 | 3.023626 |
The figures below show QTL maps for each phenotype
out <- scan1(pr.qc, gm$covar["ICI.Early.vs.PBS_ICI.Late"], Xcovar=Xcovar, model="binary")
summary_table<-data.frame(unclass(summary(operm, alpha=c(0.01, 0.05, 0.1))))
plot_lod<-function(out,map){
for (i in 1:dim(out)[2]){
#png(filename=paste0("/Users/chenm/Documents/qtl/Jai/",colnames(out)[i], "_lod.png"))
ymx <- maxlod(out) # overall maximum LOD score
plot(out, map, lodcolumn=i, col="slateblue", ylim=c(0, ymx+0.5))
legend("topright", lwd=2, colnames(out)[i], bg="gray90")
title(main = paste0(colnames(out)[i], " [positions in cM]"))
add_threshold(map, summary(operm,alpha=0.1), col = 'purple')
add_threshold(map, summary(operm, alpha=0.05), col = 'red')
add_threshold(map, summary(operm, alpha=0.01), col = 'blue')
#par(mar=c(5.1, 6.1, 1.1, 1.1))
ymx <- 11 # overall maximum LOD score
plot(out, map, lodcolumn=i, col="slateblue", ylim=c(0, ymx+0.5))
legend("topright", lwd=2, colnames(out)[i], bg="gray90")
title(main = paste0(colnames(out)[i], " [positions in cM] \n(using same scale as eoi vs ici for easier comparison)"))
add_threshold(map, summary(operm, alpha=0.1), col = 'purple')
add_threshold(map, summary(operm, alpha=0.05), col = 'red')
add_threshold(map, summary(operm, alpha=0.01), col = 'blue')
#for (j in 1: dim(summary_table)[1]){
# abline(h=summary_table[j, i],col="red")
# text(x=400, y =summary_table[j, i]+0.12, labels = paste("p=", row.names(summary_table)[j]))
#}
#dev.off()
}
}
plot_lod(out,gm$gmap)
LOD peaks
The table below shows QTL peaks associated with the phenotype. We use the 95% threshold from the permutations to find peaks.
Centimorgan (cM)
peaks <- find_peaks(out, gm$gmap, threshold=summary(operm,alpha=0.05)$A, thresholdX = summary(operm,alpha=0.05)$X, peakdrop=3, drop=1.5)
if(nrow(peaks) >0){
peaks$marker <- find_marker(gm$gmap, chr=peaks$chr,pos=peaks$pos)
names(peaks)[2] <- c("phenotype")
peaks <- peaks[-1]
rownames(peaks) <- NULL
print(kable(peaks, escape = F, align = c("cccccccc"), "html")
%>% kable_styling("striped", full_width = T)%>%
column_spec(1, bold=TRUE)
)
#plot only peak chromosomes
plot_lod_chr<-function(out,map,chrom){
for (i in 1:dim(out)[2]){
#png(filename=paste0("/Users/chenm/Documents/qtl/Jai/",colnames(out)[i], "_lod.png"))
#par(mar=c(5.1, 6.1, 1.1, 1.1))
ymx <- maxlod(out) # overall maximum LOD score
plot(out, map, chr = chrom, lodcolumn=i, col="slateblue", ylim=c(0, ymx+0.5))
#legend("topright", lwd=2, colnames(out)[i], bg="gray90")
title(main = paste0(colnames(out)[i], " - chr", chrom, " [positions in cM]"))
add_threshold(map, summary(operm,alpha=0.1), col = 'purple')
add_threshold(map, summary(operm, alpha=0.05), col = 'red')
add_threshold(map, summary(operm, alpha=0.01), col = 'blue')
#for (j in 1: dim(summary_table)[1]){
# abline(h=summary_table[j, i],col="red")
# text(x=400, y =summary_table[j, i]+0.12, labels = paste("p=", row.names(summary_table)[j]))
#}
#dev.off()
ymx <- 11
plot(out, map, chr = chrom, lodcolumn=i, col="slateblue", ylim=c(0, ymx+0.5))
#legend("topright", lwd=2, colnames(out)[i], bg="gray90")
title(main = paste0(colnames(out)[i], " - chr", chrom, " [positions in cM]\n(using same scale as eoi vs. ici for easier comparison)"))
add_threshold(map, summary(operm,alpha=0.1), col = 'purple')
add_threshold(map, summary(operm, alpha=0.05), col = 'red')
add_threshold(map, summary(operm, alpha=0.01), col = 'blue')
}
}
for(i in unique(peaks$chr)){
#for (i in 1:nrow(peaks)){
#plot_lod_chr(out,gm$gmap, peaks$chr[i])
plot_lod_chr(out,gm$gmap, i)
}
} else {
print(paste0("There are no peaks that have a LOD that reaches suggestive (p<0.05) level of ",summary(operm,alpha=0.05)$A, " [autosomes]/",summary(operm,alpha=0.05)$X, " [x-chromosome]"))
}[1] “There are no peaks that have a LOD that reaches suggestive (p<0.05) level of 3.30509555575412 [autosomes]/3.23756204890165 [x-chromosome]”
Megabase (MB)
print("peaks in MB positions")[1] “peaks in MB positions”
peaks_mba <- find_peaks(out, gm$pmap, threshold=summary(operm,alpha=0.05)$A, thresholdX = summary(operm,alpha=0.05)$X, peakdrop=3, drop=1.5)
if(nrow(peaks) >0){
peaks_mba$marker <- find_marker(gm$pmap, chr=peaks_mba$chr,pos=peaks_mba$pos)
names(peaks_mba)[2] <- c("phenotype")
peaks_mba <- peaks_mba[-1]
#peaks_mbl <- list()
##corresponding info in Mb
#for(i in 1:nrow(peaks)){
# #lodindex <- peaks$lodindex[i]
# phenotype <- peaks$phenotype[i]
# chr <- as.character(peaks$chr[i])
# lod <- peaks$lod[i]
# mark <- peaks$marker[i]
# pos <- mapdf[mapdf$marker==mark,]$pmapdf
# ci_lo <- mapdfnd$pmapdf[which(mapdfnd$gmapdf == peaks$ci_lo[i] & mapdfnd$chr == peaks$chr[i])]
# ci_hi <- mapdfnd$pmapdf[which(mapdfnd$gmapdf == peaks$ci_hi[i] & mapdfnd$chr == peaks$chr[i])]
# peaks_mb=as.data.frame(cbind(phenotype, chr, pos, lod, ci_lo, ci_hi, mark))
# names(peaks_mb)[7] <- c("marker")
# peaks_mbl[[i]] <- peaks_mb
#}
#peaks_mba2 <- do.call(rbind, peaks_mbl)
#peaks_mba2 <- as.data.frame(peaks_mba)
#peaks_mba[,c("chr", "pos", "lod", "ci_lo", "ci_hi")] <- sapply(peaks_mba[,c("chr", "pos", "lod", "ci_lo", "ci_hi")], as.numeric)
rownames(peaks_mba) <- NULL
print(kable(peaks_mba, escape = F, align = c("cccccccc"), "html")
%>% kable_styling("striped", full_width = T)%>%
column_spec(1, bold=TRUE)
)
plot_lod_chr_mb<-function(out,map,chrom){
for (i in 1:dim(out)[2]){
#png(filename=paste0("/Users/chenm/Documents/qtl/Jai/",colnames(out)[i], "_lod.png"))
#par(mar=c(5.1, 6.1, 1.1, 1.1))
ymx <- maxlod(out) # overall maximum LOD score
plot(out, map, chr = chrom, lodcolumn=i, col="slateblue", ylim=c(0, ymx+0.5))
#legend("topright", lwd=2, colnames(out)[i], bg="gray90")
title(main = paste0(colnames(out)[i], " - chr", chrom, " [positions in MB]"))
add_threshold(map, summary(operm,alpha=0.1), col = 'purple')
add_threshold(map, summary(operm, alpha=0.05), col = 'red')
add_threshold(map, summary(operm, alpha=0.01), col = 'blue')
#for (j in 1: dim(summary_table)[1]){
# abline(h=summary_table[j, i],col="red")
# text(x=400, y =summary_table[j, i]+0.12, labels = paste("p=", row.names(summary_table)[j]))
#}
#dev.off()
ymx <- 11
plot(out, map, chr = chrom, lodcolumn=i, col="slateblue", ylim=c(0, ymx+0.5))
#legend("topright", lwd=2, colnames(out)[i], bg="gray90")
title(main = paste0(colnames(out)[i], " - chr", chrom, " [positions in MB]\n(using same scale as eoi vs. ici for easier comparison)"))
add_threshold(map, summary(operm,alpha=0.1), col = 'purple')
add_threshold(map, summary(operm, alpha=0.05), col = 'red')
add_threshold(map, summary(operm, alpha=0.01), col = 'blue')
}
}
for(i in unique(peaks_mba$chr)){
#for (i in 1:nrow(peaks_mba)){
#plot_lod_chr_mb(out,gm$pmap, peaks_mba$chr[i])
plot_lod_chr_mb(out,gm$pmap,i)
}
} else {
print(paste0("There are no peaks that have a LOD that reaches suggestive (p<0.05) level of ",summary(operm,alpha=0.05)$A, " [autosomes]/",summary(operm,alpha=0.05)$X, " [x-chromosome]"))
}[1] “There are no peaks that have a LOD that reaches suggestive (p<0.05) level of 3.30509555575412 [autosomes]/3.23756204890165 [x-chromosome]”
QTL effects
For each peak LOD location we give a list of gene
query_variants <- create_variant_query_func("/Users/corneb/Documents/MyJax/CS/Projects/support.files/qtl2/cc_variants.sqlite")
query_genes <- create_gene_query_func("/Users/corneb/Documents/MyJax/CS/Projects/support.files/qtl2/mouse_genes_mgi.sqlite")
if(nrow(peaks) >0){
for (i in 1:nrow(peaks)){
#for (i in 1:1){
#Plot 1
#marker = find_marker(gm$gmap, chr=peaks$chr[i], pos=peaks$pos[i])
#gp <- g[,marker]
#gp[gp==1] <- "AA"
#gp[gp==2] <- "AB"
#gp[gp==0] <- NA
#plot_pxg(gp, gm$covar[,peaks$phenotype[i]], ylab=peaks$phenotype[i], sort=FALSE)
#title(main = paste0("chr: ", chr=peaks$chr[i],"; pos: ", peaks$pos[i], "cM /",peaks_mba$pos[i],"MB\n(",peaks$phenotype[i]," )"), line=0.2)
###dev.off()
g <- maxmarg(pr.qc, gm$gmap, chr=peaks$chr[i], pos=peaks$pos[i], return_char=TRUE)
#png(filename=paste0("/Users/chenm/Documents/qtl/Jai/","qtl_effect_", i, ".png"))
#par(mar=c(4.1, 4.1, 1.5, 0.6))
plot_pxg(g, gm$covar[,peaks$phenotype[i]], ylab=peaks$phenotype[i], sort=FALSE)
title(main = paste0("chr: ", chr=peaks$chr[i],"; pos: ", peaks$pos[i], "cM /",peaks_mba$pos[i],"MB\n(",peaks$phenotype[i]," )"), line=0.2)
##dev.off()
chr = peaks$chr[i]
# Plot 2
pr_sub <- pull_genoprobint(pr.qc, gm$gmap, chr, c(peaks$ci_lo[i], peaks$ci_hi[i]))
#coeff <- scan1coef(pr[,chr], cross$pheno[,peaks$lodcolumn[i]], addcovar = addcovar)
#coeff <- scan1coef(pr[,chr], cross$pheno[,peaks$lodcolumn[i]], Xcovar=Xcovar)
#coeff <- scan1coef(pr.qc[,chr], gm$covar[peaks$lodcolumn[i]], model="binary")
#coeff_sub <- scan1coef(pr_sub[,chr], gm$covar[peaks$lodcolumn[i]], model="binary")
blup <- scan1blup(pr.qc[,chr], gm$covar[peaks$phenotype[i]])
blup_sub <- scan1blup(pr_sub[,chr], gm$covar[peaks$phenotype[i]])
write.csv(as.data.frame(blup_sub), paste0("data/ici-early.vs.pbs.ici-late.ici-late_blup_sub_chr-",chr,"_peak.marker-",peaks$marker[i],"_lod.drop-1.5_snpsqc_5.batches.csv"), quote=F)
#plot_coef(coeff,
# gm$gmap, columns=1:2,
# bgcolor="gray95", legend="bottomleft",
# main = paste("chr", chr=peaks$chr[i],"; pos: ", peaks$pos[i], "cM /",peaks_mba$pos[i],"MB\n(",peaks$lodcolumn[i]," [scan1coeff; positions in cM])")
# )
#plot_coef(coeff_sub,
# gm$gmap, columns=1:2,
# bgcolor="gray95", legend="bottomleft",
# main = paste("chr", chr=peaks$chr[i],"; pos: ", peaks$pos[i], "cM /",peaks_mba$pos[i],"MB\n(",peaks$lodcolumn[i],"; 1.5 LOD drop interval [scan1coeff; positions in cM] ) ")
# )
plot_coef(blup,
gm$gmap, columns=1:2,
bgcolor="gray95", legend="bottomleft",
main = paste0("chr: ", chr=peaks$chr[i],"; pos: ", peaks$pos[i], "cM /",peaks_mba$pos[i],"MB\n(",peaks$phenotype[i]," [scan1blup; positions in cM])")
)
plot_coef(blup_sub,
gm$gmap, columns=1:2,
bgcolor="gray95", legend="bottomleft",
main = paste0("chr: ", chr=peaks$chr[i],"; pos: ", peaks$pos[i], "cM /",peaks_mba$pos[i],"MB\n(",peaks$phenotype[i],"; 1.5 LOD drop interval [scan1blup; positions in cM])")
)
# Plot 3
#c2effB <- scan1coef(pr.qc[,chr], gm$covar[peaks$lodcolumn[i]], model="binary", contrasts=cbind(a=c(-1, 0), d=c(0, -1)))
#c2effBb <- scan1blup(pr.qc[,chr], gm$covar[peaks$lodcolumn[i]], contrasts=cbind(a=c(-1, 0), d=c(0, -1)))
##c2effB <- scan1coef(pr[,chr], cross$pheno[,peaks$lodcolumn[i]], addcovar = addcovar, contrasts=cbind(mu=c(1,1,1), a=c(-1, 0, 1), d=c(0, 1, 0)))
##c2effB <- scan1coef(pr[,chr], cross$pheno[,peaks$lodcolumn[i]],Xcovar=Xcovar, contrasts=cbind(mu=c(1,1,1), a=c(-1, 0, 1), d=c(0, 1, 0)))
#plot(c2effB, gm$gmap[chr], columns=1:2,
# bgcolor="gray95", legend="bottomleft",
# main = paste("chr", chr=peaks$chr[i], "pos", peaks$pos[i], "(",peaks$lodcolumn[i],")")
# )
#plot(c2effBb, gm$gmap[chr], columns=1:2,
# bgcolor="gray95", legend="bottomleft",
# main = paste("chr", chr=peaks$chr[i], "pos", peaks$pos[i], "(",peaks$lodcolumn[i],")")
# )
##last_coef <- unclass(c2effB)[nrow(c2effB),2:3] # last two coefficients
##for(t in seq(along=last_coef))
## axis(side=4, at=last_coef[t], names(last_coef)[t], tick=FALSE)
#Table 1
chr = peaks_mba$chr[i]
start=as.numeric(peaks_mba$ci_lo[i])
end=as.numeric(peaks_mba$ci_hi[i])
genesgss = query_genes(chr, start, end)
write.csv(genesgss, file=paste0("data/ici-early.vs.pbs.ici-late_genes_chr-",chr,"_peak.marker-",peaks$marker[i],"_lod.drop-1.5_snpsqc_5.batches.csv"), quote=F)
rownames(genesgss) <- NULL
genesgss$strand_old = genesgss$strand
genesgss$strand[genesgss$strand=="+"] <- "positive"
genesgss$strand[genesgss$strand=="-"] <- "negative"
#genesgss <-
#table <-
#genesgss[,c("chr","type","start","stop","strand","ID","Name","Dbxref","gene_id","mgi_type","description")] %>%
#kable(escape = F,align = c("ccccccccccc")) %>%
#kable_styling("striped", full_width = T) #%>%
#cat #%>%
#column_spec(1, bold=TRUE)
#
#print(kable(genesgss[,c("chr","type","start","stop","strand","ID","Name","Dbxref","gene_id","mgi_type","description")], escape = F,align = c("ccccccccccc")))
print(kable(genesgss[,c("chr","type","start","stop","strand","ID","Name","Dbxref","gene_id","mgi_type","description")], "html") %>% kable_styling("striped", full_width = T))
}
} else {
print(paste0("There are no peaks that have a LOD that reaches suggestive (p<0.05) level of ",summary(operm,alpha=0.05)$A, " [autosomes]/",summary(operm,alpha=0.05)$X, " [x-chromosome]"))
}[1] “There are no peaks that have a LOD that reaches suggestive (p<0.05) level of 3.30509555575412 [autosomes]/3.23756204890165 [x-chromosome]”
R/qtl
scanone
gmObject of class cross2 (crosstype "bc")
Total individuals 138
No. genotyped individuals 138
No. phenotyped individuals 138
No. with both geno & pheno 138
No. phenotypes 1
No. covariates 8
No. phenotype covariates 0
No. chromosomes 20
Total markers 26548
No. markers by chr:
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
2374 2374 1623 1691 1568 1703 1464 1468 1685 974 1671 1134 1377 1411 876 751
17 18 19 X
330 770 920 384 #detach("package:qtl2", unload=TRUE)
#library(qtl)
cross <- qtl::read.cross("csv", file = "data/ici-early.vs.pbs.ici-late_gm_qtl_snpsqc_5.batches.csv",alleles=c("A","B")) --Read the following data:
138 individuals
26548 markers
3 phenotypes
--Cross type: bc cross <- qtl::jittermap(cross)
summary(cross) Backcross
No. individuals: 138
No. phenotypes: 3
Percent phenotyped: 100 100 100
No. chromosomes: 20
Autosomes: 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
X chr: X
Total markers: 26548
No. markers: 2374 2374 1623 1691 1568 1703 1464 1468 1685 974 1671
1134 1377 1411 876 751 330 770 920 384
Percent genotyped: 99.5
Genotypes (%):
Autosomes: AA:54.4 AB:45.6
X chromosome: AA:53.3 AB:46.7 cross.probs <- qtl::calc.genoprob(cross)
print("method == hk")[1] "method == hk"scanone.hk <-qtl::scanone(cross.probs, pheno.col="ICI.Early.vs.PBS_ICI.Late" , model="binary", method="hk")
operm.hk <- qtl::scanone(cross.probs, method = "hk", pheno.col="ICI.Early.vs.PBS_ICI.Late", n.perm = 10, perm.Xsp = TRUE, model="binary", verbose=FALSE)
plot(operm.hk)
print(summary(operm.hk, alpha=c(0.01, 0.05, 0.1)))Autosome LOD thresholds (10 permutations)
lod
1% 2.40
5% 2.35
10% 2.29
X chromosome LOD thresholds (186 permutations)
lod
1% 2.02
5% 1.95
10% 1.86#plot(scanone.hk, bandcol = "grey90",lty=1, cex=1, col = "steelblue")
#qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.01, col = 'blue')
#qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.05, col = 'red')
#qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.1, col = 'purple')
ymx <- maxlod(out) # overall maximum LOD score
plot(scanone.hk, bandcol = "grey90",lty=1, cex=1, col = "slateblue", ylim=c(0, ymx+0.5))
title(main = paste0(colnames(out), " [positions in cM]"))
qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.01, col = 'blue')
qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.05, col = 'red')
qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.1, col = 'purple')
ymx <- 11
plot(scanone.hk, bandcol = "grey90",lty=1, cex=1, col = "slateblue", ylim=c(0, ymx+0.5))
title(main = paste0(colnames(out), " [positions in cM]\n(using same scale as eoi vs ici for easier comparison)"))
qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.01, col = 'blue')
qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.05, col = 'red')
qtl::add.threshold(scanone.hk, perms= operm.hk, alpha=0.1, col = 'purple')
print(as.data.frame(summary(scanone.hk, perms=operm.hk, pvalues=TRUE, format="allpeaks"))) chr pos lod pval
JAX00266954 1 56.986485 0.5422253 1.0000000
UNC3037495 2 29.574388 1.7768438 0.9116914
UNC6262790 3 61.266218 0.7626221 1.0000000
UNCJPD008579 4 47.710737 0.7036325 1.0000000
JAX00584372 5 32.822667 0.7155026 1.0000000
sanger3399 6 12.622185 1.4779784 1.0000000
JAX00651930r 7 60.826242 0.6429546 1.0000000
UNC15363342 8 49.712289 0.4628329 1.0000000
UNC16706804 9 43.753042 0.3839544 1.0000000
UNCHS029502 10 69.917900 1.1417450 1.0000000
UNCHS030712 11 32.135777 1.4083023 1.0000000
UNC21550240 12 40.656904 0.3839466 1.0000000
UNC22384241 13 13.831342 0.3828726 1.0000000
UNC24913816 14 65.313402 0.3839651 1.0000000
UNCHS040394 15 24.333130 0.5373864 1.0000000
UNC27138651 16 43.246503 0.2979134 1.0000000
UNC27988525 17 26.059014 0.5045240 1.0000000
UNCHS046092 18 22.098257 0.6432228 1.0000000
UNCHS047064 19 3.995011 1.3894202 1.0000000
UNC31568433 X 77.941379 0.3180227 1.0000000print("all peaks with a p-value less or equal to 0.05 (suggestive)")[1] "all peaks with a p-value less or equal to 0.05 (suggestive)"print(as.data.frame(summary(scanone.hk, perms=operm.hk, alpha=0.05, pvalues=TRUE, format="allpeaks")))[1] chr pos lod
<0 rows> (or 0-length row.names)#print("method == ehk")
#scanone.ehk <-qtl::scanone(cross.probs, pheno.col="ICI.Early.vs.PBS_ICI.Late" , model="binary", method="ehk")
#operm.ehk <- qtl::scanone(cross.probs, method = "ehk", pheno.col="ICI.Early.vs.PBS_ICI.Late", n.perm = 1000, perm.Xsp = TRUE, model="binary", verbose=FALSE)
#plot(operm.ehk)
#print(summary(operm.ehk, alpha=c(0.01, 0.05, 0.1)))
#plot(scanone.ehk, bandcol = "grey90",lty=1, cex=1, col = "steelblue")
#qtl::add.threshold(scanone.ehk, perms= operm.ehk, alpha=0.01, col = 'blue')
#qtl::add.threshold(scanone.ehk, perms= operm.ehk, alpha=0.05, col = 'red')
#qtl::add.threshold(scanone.ehk, perms= operm.ehk, alpha=0.1, col = 'purple')
#print(as.data.frame(summary(scanone.ehk)))
#print(as.data.frame(summary(scanone.ehk, perms=operm.ehk, alpha=0.05, pvalues=TRUE, format="allpeaks")))
R version 3.6.2 (2019-12-12)
Platform: x86_64-apple-darwin15.6.0 (64-bit)
Running under: macOS Catalina 10.15.7
Matrix products: default
BLAS: /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libRblas.0.dylib
LAPACK: /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libRlapack.dylib
locale:
[1] en_AU.UTF-8/en_AU.UTF-8/en_AU.UTF-8/C/en_AU.UTF-8/en_AU.UTF-8
attached base packages:
[1] stats graphics grDevices utils datasets methods base
other attached packages:
[1] abind_1.4-5 qtl2_0.22 reshape2_1.4.4 ggplot2_3.3.5
[5] tibble_3.1.2 psych_2.0.7 readxl_1.3.1 cluster_2.1.0
[9] dplyr_1.0.8 optparse_1.6.6 rhdf5_2.28.1 mclust_5.4.6
[13] tidyr_1.0.2 data.table_1.14.0 knitr_1.33 kableExtra_1.1.0
[17] workflowr_1.6.2
loaded via a namespace (and not attached):
[1] httr_1.4.1 bit64_4.0.5 viridisLite_0.4.0 assertthat_0.2.1
[5] highr_0.9 blob_1.2.1 cellranger_1.1.0 yaml_2.2.1
[9] pillar_1.6.1 RSQLite_2.2.7 backports_1.2.1 lattice_0.20-38
[13] glue_1.4.2 digest_0.6.27 promises_1.1.0 rvest_0.3.5
[17] colorspace_2.0-2 htmltools_0.5.1.1 httpuv_1.5.2 plyr_1.8.6
[21] pkgconfig_2.0.3 purrr_0.3.4 scales_1.1.1 webshot_0.5.2
[25] qtl_1.46-2 getopt_1.20.3 later_1.0.0 git2r_0.26.1
[29] generics_0.0.2 ellipsis_0.3.2 cachem_1.0.5 withr_2.4.2
[33] cli_3.0.0 mnormt_1.5-7 magrittr_2.0.1 crayon_1.4.1
[37] memoise_2.0.0 evaluate_0.14 fs_1.4.1 fansi_0.5.0
[41] nlme_3.1-142 xml2_1.3.1 tools_3.6.2 hms_0.5.3
[45] lifecycle_1.0.1 stringr_1.4.0 Rhdf5lib_1.6.3 munsell_0.5.0
[49] compiler_3.6.2 rlang_1.0.2 grid_3.6.2 rstudioapi_0.13
[53] rmarkdown_2.1 gtable_0.3.0 DBI_1.1.1 R6_2.5.0
[57] fastmap_1.1.0 bit_4.0.4 utf8_1.2.1 rprojroot_1.3-2
[61] readr_1.3.1 stringi_1.7.2 parallel_3.6.2 Rcpp_1.0.7
[65] vctrs_0.3.8 tidyselect_1.1.2 xfun_0.24