#'@title Main function to perform Fast functional association analysis based on wavelet
#'@description Perform a screening of a signal for a given phenotype and a specified level of resolution
#'@param Y phenotype vector, has to be numeric. For case-control code, it as 0 and 1.
#'@param signal signals matrix (either data.frame or numeric matrix). Lines=SNPs in increasing order in terms of base pair, columns=individuals. No missing values are allowed.
#'@param pos vector of spatial position of the variables. It has to be in the same order and length as the signal line order/length.
#'@param confounder the confounding matrix with the same sample order as Y. The intercept should not be included. If missing will generate an intercept matrix.
#'@param lev_res the maximum level of resolution needed
#'@param sigma_b the parameter of the NIG prior used for the Bayes Factor computation. We advised setting this value between 0.1 and 0.2
#'@param para logical parameter for parallelization, if not specified, set at FALSE.
#'@details The ffw function computes the Likelihood ratio used for testing the significance of a genetic region. It computes the proportion of wavelet coefficients associated with the resolution level and the Bayes factor used for this estimation.
#'@return A named vector. First, position the estimated value of the Lambda statistics, then the proportion of association per level of resolution, then the computed Bayes Factor per wavelet coefficient.
#'@references Shim and Stephens, Wavelet-based genetic association analysis of functional phenotypes arising from high-throughput sequencing assays, The Annals of Applied Statistics, 2015, Vol. 9, No. 2, 665–686
#'@export
#'@examples \dontrun{
#'
#'
#'set.seed(66)
#'#########################################
#'#Generate a randomly sample signal size=1Mb
#'#########################################
#'
#'#5000 Randomly choosen pos
#'my_pos <- sort(sample(1:1000000, size=5000,replace = FALSE))
#'#############################
#'#Three different bump signals
#'#############################
#'my_functions <-data.frame(f0 = c(rep(0,400000),rep(0,200000),rep(0,400000)),
#' f1 = c(rep(0,400000),rep(1,200000),rep(0,400000)) ,
#' f2=c(rep(0,400000),rep(2,200000),rep(0,400000)))
#'
#'
#'library(gridExtra)
#'###########################
#'#Minor allele frequency 30%
#'###########################
#'MAF=0.3
#'sampl_schem <- c((1-MAF)^2,2*MAF*(1-MAF),MAF^2)
#'#######################################
#'#sampling at Hardy Weinberg equilibrium
#'#######################################
#'#Assigning class
#'
#'#sample size =4000
#'n_size=4000
#'type_fn <-sample(0:2,replace = TRUE,size=n_size, prob= sampl_schem )
#'
#'
#'signals <- matrix(my_functions[my_pos,2 ], ncol=1 ) %*%t(matrix(type_fn,ncol=1))
#'#dim(signals)= nSNP, nind
#'
#'###############################################################
#'#Generate a phenotype with variance explained by signals 0.5%
#'###############################################################
#'varexp=0.005
#'var_noise <- (1-varexp)*var(sample(0:2,replace = TRUE,size=10000,
#' prob=sampl_schem ))/varexp
#'Y <- rnorm(n=n_size,sd=sqrt(var_noise)) +type_fn
#'df <- data.frame(y=Y,signals =factor(type_fn))
#'P1 <- ggplot(df,aes(y=y,x=signals))+
#' geom_boxplot()+
#' xlab("Type of signals")+
#' theme(axis.text=element_text(size=12),
#' axis.title=element_text(size=14,face="bold"))+
#' ylab("Simulated Phenotype")+
#' theme_bw()+
#' ggtitle("Variation of the phenotype\ndepending of the signals, \nVariance explained =0.5%")
#'
#'df <- data.frame(pos= rep(my_pos,3),y=c(my_functions[my_pos,1],my_functions[my_pos,2],my_functions[my_pos,3]),
#' mycol = factor(c(rep("f0",length(my_pos)),rep("f1",length(my_pos)),rep("f2",length(my_pos))) ) )
#'
#'P2 <- ggplot(df,aes(y=y,x=pos,color=mycol))+
#' geom_point(size=1)+
#' xlab("Base pair")+
#' ylab("Number of variants")+
#' theme_bw()+
#' theme(legend.title=element_blank())+
#' ggtitle("Three different kind of signals signal")
#'
#'grid.arrange(P1,P2,ncol=2)
#'
#'##################
#'#Screening
#'##################
#'res <- ffw( Y,signal=signals,pos=my_pos,
#' lev_res=6,sigma_b = 0.2)
#'# or:
#'signals_df <- as.data.frame(signals)
#'res <- ffw( Y,signal=signals_df,pos=my_pos,
#' lev_res=6,sigma_b = 0.2)
#'#value of the test statistic
#'res["Lambda"]
#'#############
#'#Significance
#'#############
#'
#'#Estimation of the Bayes factor lambda_1 distribution parameter
#'#Take a bit of time
#'lambda <- get_lambda1(Y,sigma_b = 0.2)
#'#Simulation of the test statistics under the null distribution of
#'# the Bayes factor
#'Sim_gam <- Simu_Lambda_null(nsimu=10000, lambda=lambda,lev_res = 6)
#'val <- res["Lambda"]
#'
#'#Via Simulation
#'
#'pval <-c(length(Sim_gam[which(Sim_gam>val)])+1)/(length(Sim_gam)+1)
#'pval
#'#'pval
#'
#'
#'##############
#'#Visualisation
#'##############
#'pos <- c(min(my_pos),max(my_pos))
#'plot_ffw(res=res,pos=pos,lev_res=6)
#'
#'
#'}
ffw <- function(Y,signal,pos,confounder,lev_res,sigma_b,para=FALSE,betas=FALSE)
{
Y <- as.vector(Y)
# INPUT CHECKS
print("Input dimensions:")
if(!is.numeric(Y) || length(Y)==0){
stop("ERROR: Y is not a numeric vector")
} else {
print(sprintf("%i phenotypes detected", length(Y)))
if(all(Y %in% c(0,1))){
print("Binary phenotype detected")
} else if(!is.vector(Y)){
stop("ERROR: Y is not a vector. Multi-phenotype analysis not implemented yet.")
} else {
print("Continuous phenotype detected")
}
}
if(missing(betas)) {
betas <- FALSE
}
# Writing the design matrix
if(missing(confounder)) {
print("no covariates provided, using intercept only")
confounder <- data.frame(confounding=rep(1,length(Y)) )
} else if(nrow(confounder)!=length(Y)) {
stop("ERROR: number of samples in Y and confounder does not match")
} else {
print(sprintf("%i covariates for %i samples detected", ncol(confounder), nrow(confounder)))
confounder <- cbind(rep(1,length(Y)),confounder)
}
# Check signals matrix
if(is.data.frame(signal)){
print("Converting signals data to matrix")
signal <- as.matrix(signal)
}
if(missing(signal) || !is.numeric(signal)){
stop("ERROR: signals matrix missing or not numeric")
} else if(ncol(signal)!=length(Y)){
stop("ERROR: number of samples in Y and signal does not match")
} else {
print(sprintf("%i SNPs for %i samples detected", nrow(signal), ncol(signal)))
}
# Check position vector
if(!is.numeric(pos) || !is.vector(pos)){
stop("ERROR: must provide numeric position vector")
} else {
print(sprintf("positions for %i SNPs read", length(pos)))
}
# Clean missing samples from all inputs
keepY <- complete.cases(Y)
keepC <- complete.cases(confounder)
keepGT <- complete.cases(t(signal))
nonmissing_index <- which(keepGT & keepY & keepC)
if(length(nonmissing_index) != length(Y)){
print(sprintf("Warning: %i individuals will be removed due to missingness",
length(Y) - length(nonmissing_index)))
}
Y <- Y[nonmissing_index]
confounder <- confounder[nonmissing_index,]
signal <- signal[,nonmissing_index]
sigma_b <- 10*sigma_b
print(paste("N individuals analysed = ", dim(signal)[2],
", N SNPs analysed = ",dim(signal)[1]))
if(is.null(dim(signal)) || dim(signal)[1] < 2^lev_res || dim(signal)[2] < 2){
print("Warning: not enough signals remaining, returning empty output")
# Naming the output
names_BF <- c("BF_0_0")
for(i in 1:lev_res){
for (j in 1:(2^i)){
names_BF <- c(names_BF,paste("BF",i,j,sep = "_"))
}
}
out = rep(NA, 1+lev_res+1+length(names_BF))
names(out) <- c("Lambda", paste("pi",0:lev_res, sep = "_"), names_BF)
return(out)
}
####################################
#Redefinition of the needed function
####################################
n_coef_wc <- function(lev_res)
{
temp <- c()
for(i in 0:lev_res)
{
temp <- c(temp,2^i)
}
sum(temp)
}
#Quantile transform to prevent for non normaliy distrib WCs
Quantile_transform <- function(x)
{
.ex.seed <- exists(".Random.seed")
if(.ex.seed) .oldseed <- .Random.seed
set.seed(666)
if(.ex.seed) on.exit(.Random.seed <<- .oldseed)
x.rank = rank(x, ties.method="random")
#x.rank = rank(x, ties.method="average")
return(qqnorm(x.rank,plot.it = F)$x)
}
#Estimation of Lambda
Lambda_stat <- function (my_pi, my_bayes)
{
# vector: pi1 pi2 pi2 pi3 pi3 pi3 pi3...
my_pi_vec = rep(my_pi, 2^(1:length(my_pi)-1))
coefs = 1-my_pi_vec + my_pi_vec * my_bayes[1:(2^length(my_pi)-1)]
prod(coefs)
}
sumlog <- function (A1, A2)
{
if(A1 > A2){
res = A1 + log(1 + exp(A2 - A1))
}else{
res = A2 + log(exp(A1 - A2) + 1)
}
return(res)
}
max_EM_Lambda <- function(my_bayes)
{
niter=10000
epsilon <- 10^-4
p_vec <- c()
for(gi in 0: lev_res)
{
# EM algorithm for each group separately
N_obllikli = 0
logpi = log(0.5)
pi <- 0.5
log1pi = logpi
pp = 0
logPiBF = log(my_bayes[(2^gi):(2^(gi+1)-1)]) + logpi
logden <- c()
for (i in 1:length(logPiBF))
{
logden[i] <- sumlog(logPiBF[i],log1pi)
}
pp = pp+sum(exp(logPiBF - logden))
N_obllikli = sum(logden)
O_obllikli = N_obllikli
for(iter in 0:niter){
pi = pp/(2^(gi))
logpi = log(pi)
log1pi = log(1-pi)
logPiBF = log(my_bayes[(2^gi):(2^(gi+1)-1)]) + logpi
logden <- c()
for (i in 1:length(logPiBF))
{
logden[i] <- sumlog(logPiBF[i],log1pi)
}
pp=0
pp = pp+sum(exp(logPiBF - logden))
N_obllikli = sum(logden)
diff = abs(N_obllikli - O_obllikli)
if(diff < epsilon){
break
}else{
O_obllikli = N_obllikli
}
}
p_vec <-c(p_vec,pi)
}
return(p_vec)
}
###############
#Paralelisation
###############
if(para==TRUE)
{
cl <-makeCluster(detectCores(all.tests=TRUE)-1, type = "SOCK")
}
###################
#Wavelet processing
###################
print("Wavelet processing")
Time01 <- (pos- min(pos))/(max(pos)-min(pos))
my_wavproc <- function(y)
{
#Kovac and Silvermann 2000
mygrid <- wavethresh::makegrid(t=Time01,y=y)
LDIRWD <- irregwd(mygrid,filter.number=1)
class(LDIRWD) <- "wd"
#Thresholding here
LDIRWD <- threshold(LDIRWD,policy = "universal",type="hard",
dev = madmad,levels = 1:(LDIRWD$nlevels-1))
res <- c()
for(i in 0: lev_res){
res <- c(res, accessD( LDIRWD,lev = i) )
}
return(res)
}
if(para==TRUE)
{
clusterExport(cl,"irregwd")
clusterExport(cl,"threshold")
clusterExport(cl,"madmad")
clusterExport(cl,"accessD")
clusterExport(cl,"accessC")
clusterExport(cl,"my_wavproc")
Gen_W_trans <- snow::parApply(cl,signal,2,my_wavproc)
}
else{
Gen_W_trans <- apply(signal,2,my_wavproc)
}
#Quantile transform for non normal WCs for every scale location
Gen_W_trans = apply(Gen_W_trans, 1, Quantile_transform)
##########
#Modeling
##########
print("Computing Bayes Factors")
W <- as.matrix(confounder, ncol=ncol(confounder))
n = nrow(W)
q = ncol(W)
# L <- as.matrix(Y , ncol=ncol(Y)) #reversed regression
L <- as.matrix(Y,ncol=1)
p = 1
PW = diag(n) - W %*% solve(t(W) %*% W) %*% t(W)
X = PW %*% L
HB = X %*% solve(t(X) %*% X + diag(1/sigma_b/sigma_b,p)) %*% t(X)
delta = svd(X)$d
lambda = delta^2 / (delta^2 + 1/sigma_b/sigma_b)
log.T = sum(log(1-lambda))/2
my_bf <- function( y ){
y <- as.matrix(y,ncol=1)
log.R = -0.5*n*log(1 - (t(y) %*% HB %*% y) / (t(y) %*% PW %*% y ))
bf = exp(log.T + log.R)
return(c(bf))
}
if(para==TRUE)
{
clusterExport(cl,"log.T")
clusterExport(cl,"sigma_b")
clusterExport(cl,"my_bf")
my_bayes <- snow::parApply(cl,Gen_W_trans, 2, my_bf )
}
else{
my_bayes <- apply(Gen_W_trans, 2, my_bf )
}
if(betas ==TRUE)
{
print("Computing Beta values")
betas_f <- function(y)
{
confounder <- data.frame(confounder)
pc <- dim(confounder)[2]
Dmat <- cbind(confounder,Y)
Dmat <- as.matrix(Dmat)
res <- solve(t(Dmat) %*% Dmat + diag(1/sigma_b/sigma_b,dim(Dmat)[2])) %*% t(Dmat)%*% y
index <- pc+1
return(res[index,1])
}
my_betas <- apply(Gen_W_trans, 2, betas_f )
}
#################
#Estimation Lambda
#################
print("Post-processing")
my_pis <- max_EM_Lambda(my_bayes = my_bayes)
trueLambda <- Lambda_stat(my_pi = my_pis,my_bayes = my_bayes)
if(betas ==FALSE)
{
out <- c(trueLambda,my_pis,my_bayes)
}
else
{
out <- c(trueLambda,my_pis,my_bayes,my_betas)
}
#Naming the output
if(betas ==FALSE)
{
names_BF <- c("BF_0_0")
for(i in 1:lev_res)
{
for (j in 1:(2^i))
{
names_BF <- c(names_BF,paste("BF",i,j,sep = "_"))
}
}
names(out) <- c("Lambda",
paste("pi",0:lev_res, sep = "_"),
names_BF)
}
else
{
names_BF <- c("BF_0_0")
for(i in 1:lev_res)
{
for (j in 1:(2^i))
{
names_BF <- c(names_BF,paste("BF",i,j,sep = "_"))
}
}
names_Betas <- c("Beta_0_0")
for(i in 1:lev_res)
{
for (j in 1:(2^i))
{
names_Betas <- c(names_Betas,paste("Beta",i,j,sep = "_"))
}
}
names(out) <- c("Lambda",
paste("pi",0:lev_res, sep = "_"),
names_BF,names_Betas)
}
if(para==TRUE)
{
stopCluster(cl)
}
return(out)
}
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