NSANormalization: The NSANormalization class

In NSA: Post-normalization of total copy numbers

Description

Package: NSA
Class NSANormalization

Object
~~|
~~+--NSANormalization

Directly known subclasses:

public static class NSANormalization
extends Object

This class represents the NSA normalization method [1], which looks for normal regions within the tumoral samples.

Usage

NSANormalization(data=NULL, tags="*", ...)

Arguments

data	A named list with data set named "total" and "fracB" where the former should be of class AromaUnitTotalCnBinarySet and the latter of class AromaUnitFracBCnBinarySet. The two data sets must be for the same chip type, have the same number of samples and the same sample names.
tags	Tags added to the output data sets.
...	Not used.

Details

...

Fields and Methods

Methods:

	findArraysTodo	-
	getDataSets	-
	getFullName	-
	getName	-
	getOutputDataSets	-
	getPath	-
	getRootPath	-
	getTags	-
	nbrOfFiles	-
	process	Finds normal regions within tumoral samples.
	setTags	-

Methods inherited from Object:
asThis, $, $<-, [[, [[<-, as.character, attach, attachLocally, clearCache, clone, detach, equals, extend, finalize, gc, getEnvironment, getFields, getInstantiationTime, getStaticInstance, hasField, hashCode, ll, load, objectSize, print, registerFinalizer, save

References

[1] ...

See Also

Low-level versions of the NSA normalization method is available via the NSAByTotalAndFracB.matrix() methods.

Examples

## Not run: 
  
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# CRMAv2 - Preprocess raw Affymetrix data
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
library("aroma.affymetrix");  # Needed for CRMAv2
#library("calmate");
library(MASS)
source("F:/MOrtiz/curro/Aroma/calmate/R/CalMaTeNormalization.R")
source("F:/MOrtiz/curro/Aroma/calmate/R/calmateByTotalAndFracB.R")
source("F:/MOrtiz/curro/Aroma/calmate/R/calmateByThetaAB.R")
source("F:/MOrtiz/curro/Aroma/calmate/R/fitCalMaTe.R")
source("F:/MOrtiz/curro/Aroma/calmate/R/fitCalMaTeCNprobes.R")
source("F:/MOrtiz/curro/Aroma/calmate/R/thetaAB2TotalAndFracB.R")

source("F:/MOrtiz/curro/Aroma/NSA/R/NSANormalization.R")
source("F:/MOrtiz/curro/Aroma/NSA/R/NSAByTotalAndFracB.R")
source("F:/MOrtiz/curro/Aroma/NSA/R/fitNSA.R")

source("F:/MOrtiz/curro/Aroma/NSA/R/SNPsNormalization.R")
source("F:/MOrtiz/curro/Aroma/NSA/R/SNPsNByTotalAndFracB.R")
source("F:/MOrtiz/curro/Aroma/NSA/R/fitSNPsN.R")

source("F:/MOrtiz/curro/Aroma/NSA/R/SampleNormalization.R")
source("F:/MOrtiz/curro/Aroma/NSA/R/SampleNByTotalAndFracB.R")
source("F:/MOrtiz/curro/Aroma/NSA/R/fitSample.R")
library("DNAcopy");

setwd("I:/aroma")

dataSet <- "breast cancer";
dataSet <- "GSE12702-prostateCancerPaired"
dataSet <- "GSE12702-prostateCancer"
dataSet <- "GSE14996,testSet"

chipType <- "Mapping250K_Nsp";
#chipType <- "GenomeWideSNP_6"
dsList <- doCRMAv2(dataSet, chipType=chipType, combineAlleles=FALSE,
                                             plm="RmaCnPlm", verbose=-10);
print(dsList);


# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# CalMaTe - Post-normalization of ASCNs estimates
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
asn <- CalMaTeNormalization(dsList);
print(asn);

# For speed issues, we will here only process loci on Chromosome 17.
chr <- 22;
ugp <- getAromaUgpFile(dsList$total);
units <- getUnitsOnChromosome(ugp, chr);

dsNList <- process(asn, units=units, verbose=verbose);
#dsNList <- process(asn, references = seq(2,40,2), verbose=verbose);
#dsNList <- process(asn, verbose=verbose);
print(dsNList);

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# NSA - Finding normal regions
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

asnN <- NSANormalization(dsNList);
print(asnN);

dsNNList <- process(asnN, verbose=verbose);
print(dsNNList);

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# NSA - SNPs Normalization
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

asnNsnps <- SNPsNormalization(dsNList);
print(asnNsnps);

dsNNListSNPs <- process(asnNsnps, references = dsNNList, verbose=verbose);
print(dsNNListSNPs);

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# NSA - Sample Normalization
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

asnNsample <- SampleNormalization(dsNNListSNPs);
print(asnNsample);

dsNNListSample <- process(asnNsample, references = dsNNList, verbose=verbose);
print(dsNNListSample);

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# NSA - SNPs Normalization
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

asnNsnps <- SNPsNormalization(dsNNListSample);
print(asnNsnps);

dsListSNPs <- process(asnNsnps, references = dsNNList, verbose=verbose);

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# NSA - Sample Normalization
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

asnNsample <- SampleNormalization(dsListSNPs);
print(asnNsample);

dsListSample <- process(asnNsample, references = dsNNList, verbose=verbose);
print(dsNNListSample);

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# CalMaTe - sigma delta validation  (for CRMAv2)
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

# Alt 1: Calculate CN ratios where the reference is a pool of specific references samples

references <- c(1:20);

dsR <- extract(dsList$total, references);
dfR <- getAverageFile(dsR);
cnm <- CopyNumberChromosomalModel(dsList$total, dfR);

# Alt 2: Calculate CN ratios where the reference is the pool of all samples
cnm <- CopyNumberChromosomalModel(dsList$total);
print(cnm);

# Extract CN ratios - C=theta/thetaR
cn <- extractRawCopyNumbers(cnm, array=ii, chromosome=chr, logBase=NULL);
cn$y <- 2*cn$y;
print(cn);

# As a data frame
#data <- as.data.frame(cn);

# Estimate std dev (via first-order variance estimator)
sigma <- estimateStandardDeviation(cn);
print(sigma);

# Plot
x11();plot(cn, ylim=c(0,6));
sigmaStr <- sprintf("%.3f", sigma);
stext(side=3, pos=0.5, line=-1, substitute(hat(sigma)[Delta]==x, list(x=sigmaStr)));

# Smooth (bins of 1.0Mb)
cnS <- binnedSmoothing(cn, by=1.0e6);
print(cnS);
points(cnS, col="red");
lines(cnS, col="red");

# Segmentation
fit <- segmentByCBS(cn);
cnr <- extractCopyNumberRegions(fit);
print(cnr);
drawLevels(cnr, col="blue", lwd=3);

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# CalMaTe - sigma delta validation  (for CalMaTe)
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

# Alt 1: Calculate CN ratios where the reference is a pool of specific references samples

references <- c(1:24)

# Alt 2: Calculate CN ratios where the reference is the pool of all samples
#cnm <- CopyNumberChromosomalModel(dsNNListSample$total);

#cnm <- CopyNumberChromosomalModel(dsNList$total);
#print(cnm);

# Extract CN ratios - C=theta/thetaR
#cn <- extractRawCopyNumbers(cnm, array=ii, chromosome=chr, logBase=NULL);

dataNSA <- extractMatrix(dsNList$total,units = units);
cnNSA <- cn;
cnNSA$y <- dataNSA[,ii];
cn <- cnNSA;


# As a data frame
data <- as.data.frame(cn);

# Estimate std dev (via first-order variance estimator)
sigma <- estimateStandardDeviation(cn);
print(sigma);

# Plot
x11();plot(cn, ylim=c(0,6));
sigmaStr <- sprintf("%.3f", sigma);
stext(side=3, pos=0.5, line=-1, substitute(hat(sigma)[Delta]==x, list(x=sigmaStr)));
                                                       
# Smooth (bins of 1.0Mb)
cnS <- binnedSmoothing(cn, by=1.0e6);
print(cnS);
points(cnS, col="red");
lines(cnS, col="red");

# Segmentation
fit <- segmentByCBS(cn);
cnr <- extractCopyNumberRegions(fit);
print(cnr);
drawLevels(cnr, col="blue", lwd=3);


#You can obtain the above 'cn' manually as:

dsR <- extract(dsList$total, references);
dfR <- getAverageFile(dsR);

df <- getFile(dsList$total, ii);
theta <- extractRawCopyNumbers(df, chromosome=chr);
print(theta);
thetaR <- extractRawCopyNumbers(dfR, chromosome=chr);
print(thetaR);
cn <- clone(theta);
cn <- divideBy(cn, thetaR);
cn$y <- 2*cn$y;

#sigma delta
mad(diff(cn$y))

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Calculating the differences between CN and SNP probes
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -


cnCNP <- RawCopyNumbers(...);
cnSNP <- RawCopyNumbers(...);

xRange <- range(xRange(cnCNP), xRange(cnSNP));

by <- 50e3; # 50kb bins; you may want to try with other amounts of smoothing xOut <- seq(from=xRange[1], to=xRange[2], by=by);

cnCNPS <- binnedSmoothing(cnCNP, xOut=xOut); cnSNPS <- binnedSmoothing(cnSNP, xOut=xOut);

Clim <- c(0,5);
plot(cnCNPS, ylim=Clim);
points(cnSNPS, col="blue");

plot(getSignals(cnCNPS), getSignals(cnSNPS), xlim=Clim, ylim=Clim); abline(a=0, b=1, col="red", lwd=2);


# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Plot allele B fractions (before and after calmate)
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Sample #1 and Chromosome 17
ii <- 1;
chr <- 17;
df <- getFile(dsList$fracB, ii);
dfN <- getFile(dsNList$fracB, ii);

beta <- extractRawAlleleBFractions(df, chromosome=chr);
betaN <- extractRawAlleleBFractions(dfN, chromosome=chr);
x11();
subplots(2, ncol=1);
plot(beta);
title(sprintf("%s", getName(beta)));
plot(betaN);
title(sprintf("%s (CalMaTe)", getName(betaN)));


for(ii in 1:24){
  df <- getFile(dsList$total, ii);
  CN <- extractRawCopyNumbers(df, chromosome=chr);
  if(ii == 1){
    aux <- CN$y;
  }else{
    aux <- aux + CN$y;
  }
}
ref <- aux/24;

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Plot copy numbers (before and after calmate)
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Sample #1 and Chromosome 17
ii <- 3;
chr <- 1;

df <- getFile(dsList$total, ii);
#dfN <- getFile(dsNList$total, ii);

#units <- getUnitsOnChromosome(gi, ii);
#pos <- getPositions(gi, units = units);
#units <- units[order(pos)];

CN <- extractRawCopyNumbers(df, chromosome=chr);
#CNN <- extractRawCopyNumbers(dfN, chromosome=chr);
#CNN <- extractMatrix(dsNList$total, units = units);
#CNN <- CNN[,ii];


x11();
subplots(2, ncol=1);
plot(CN$x, log2(CN$y));
title(sprintf("%s", getName(CN)));
plot(CNN);
title(sprintf("%s (CalMaTe)", getName(CNN)));

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Plot Normal Regions
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Sample #3 and Chromosome 6
ii <- 1;
chr <- 6;
dfNN <- getFile(dsNNList$normalReg, ii);

betaNN <- extractRawAlleleBFractions(dfNN, chromosome=chr);

plot(betaNN);
title(sprintf("%s (NSA)", getName(betaNN)));

# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Plot Normalized by SNP data
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Sample #9 and Chromosome 8
ii <- 9;
chr <- 8;
dfN <- getFile(dsNNListSNPs$fracB, ii);
fracBN <- extractRawAlleleBFractions(dfN, chromosome=chr);

dfN <- getFile(dsNNListSNPs$total, ii);
totalN <- extractRawCopyNumbers(dfN, chromosome=chr);

x11();
subplots(2, ncol=1);
plot(fracBN);
title(sprintf("%s", getName(fracBN)));
plot(totalN);
title(sprintf("%s ", getName(totalN)));


# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Plot Normalized by Sample data
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# Sample #3 and Chromosome 6
ii <- 9;
chr <- 8;

dfC <- getFile(dsNList$total, ii);
CNC <- extractRawCopyNumbers(dfC, chromosome=chr);

dfN <- getFile(dsNNListSample$fracB, ii);
fracBN <- extractRawAlleleBFractions(dfN, chromosome=chr);

dfN <- getFile(dsNNListSample$total, ii);
totalN <- extractRawCopyNumbers(dfN, chromosome=chr);

x11();
subplots(2, ncol=1);
plot(fracBN);
title(sprintf("%s", getName(fracBN)));
plot(totalN$x, 2^totalN$y);
title(sprintf("%s ", getName(totalN)));

x11();
plot(CNC);
points(totalN, col="green");

 
## End(Not run)

NSA documentation built on May 2, 2019, 4:58 p.m.