glad {GLAD} | R Documentation

## Analysis of array CGH data

### Description

This function allows the detection of breakpoints in genomic profiles obtained by array CGH technology and affects a status (gain, normal or lost) to each clone.

### Usage

    ## S3 method for class 'profileCGH'
    glad(profileCGH, mediancenter=FALSE,
                    smoothfunc="lawsglad", bandwidth=10, round=1.5,
                    model="Gaussian", lkern="Exponential", qlambda=0.999,
                    base=FALSE, sigma,
                    lambdabreak=8, lambdacluster=8, lambdaclusterGen=40,
                    type="tricubic", param=c(d=6),
                    alpha=0.001, msize=5,
                    method="centroid", nmax=8, assignGNLOut=TRUE,
                    breaksFdrQ = 0.0001, haarStartLevel = 1, haarEndLevel = 5,
                    verbose=FALSE, ...)

### Arguments

`profileCGH` | Object of class `profileCGH`
`mediancenter` | If `TRUE`, LogRatio are centered on their median.
`smoothfunc` | Type of algorithm used to smooth `LogRatio` by a piecewise constant function. Choose either `lawsglad`, `haarseg`, `aws` or `laws` in aws package.
`bandwidth` | Set the maximal bandwidth `hmax` in the `aws` or `laws` functions in aws package. For example, if `bandwidth=10` then the `hmax` value is set to 10*X_N where X_N is the position of the last clone.
`round` | The smoothing results are rounded or not depending on the `round` argument. The `round` value is passed to the argument `digits` of the `round` function.
`model` | Determines the distribution type of the LogRatio. Keep always the model as "Gaussian" (see `laws` in aws package).
`lkern` | Determines the location kernel to be used (see `aws` or `laws` in aws package).
`qlambda` | Determines the scale parameter for the stochastic penalty (see `aws` or `laws` in aws package)
`base` | If `TRUE`, the position of clone is the physical position on the chromosome, otherwise the rank position is used.
`sigma` | Value to be passed to either argument `sigma2` of`aws` function or `shape` of `laws` (see aws package). If `NULL`, sigma is calculated from the data.
`lambdabreak` | Penalty term (λ') used during the Optimization of the number of breakpoints step.
`lambdacluster` | Penalty term (λ*) used during the MSHR clustering by chromosome step.
`lambdaclusterGen` | Penalty term (λ*) used during the HCSR clustering throughout the genome step.
`type` | Type of kernel function used in the penalty term during the Optimization of the number of breakpoints step, the MSHR clustering by chromosome step and the HCSR clustering throughout the genome step.
`param` | Parameter of kernel used in the penalty term.
`alpha` | Risk alpha used for the Outlier detection step.
`msize` | The outliers MAD are calculated on regions with a cardinality greater or equal to msize.
`method` | The agglomeration method to be used during the MSHR clustering by chromosome and the HCSR clustering throughout the genome clustering steps.
`nmax` | Maximum number of clusters (N*max) allowed during the the MSHR clustering by chromosome and the HCSR clustering throughout the genome clustering steps.
`assignGNLOut` | If `FALSE` the status (gain/normal/loss) is not assigned for outliers.
`breaksFdrQ` | breaksFdrQ for HaarSeg algorithm.
`haarStartLevel` | haarStartLevel for HaarSeg algorithm.
`haarEndLevel` | for HaarSeg algorithm.
`verbose` | If `TRUE` some information are printed
`...` | ...

### Details

The function `glad` implements the methodology which is described in the article: Analysis of array CGH data: from signal ratio to gain and loss of DNA regions (HupÃ© et al., Bioinformatics, 2004).

The principles of the GLAD algorithm: First, the detection of breakpoints is based on the estimation of a piecewise constant function with the Adaptive Weights Smoothing (AWS) procedure (Polzehl and Spokoiny, 2002). Alternatively, it is possible to use the HaarSeg algorithm (Ben-Yaacov and Eldar, Bioinformatics, 2008). Then, a procedure based on penalyzed maximum likelihood optimizes the number of breakpoints and removes the undesirable breakpoints. Finally, based on the regions previously identified, a two-step unsupervised classification (MSHR clustering by chromosome and the HCSR clustering throughout the genome) with model selection criteria allows a status to be assigned for each region (gain, loss or normal).

Main parameters to be tuned:

`qlambda` | if you want the smoothing to fit some very local effect, choose a smaller `qlambda`.
`bandwidth` | choose a bandwidth not to small otherwise you will have a lot of little discontinuities.
`lambdabreak` | The higher the parameter is, the higher the number of undesirable breakpoints is.
`lambdacluster` | The higher the parameter is, the higher is the number of the regions within a chromosome which belong to the same cluster.
`lambdaclusterGen` | More the parameter is high more the regions over the whole genome are supposed to belong to the same cluster.

### Value

`` | An object of class "profileCGH" with the following attributes:
`profileValues:` | a data.frame with the following added information: SmoothingThe smoothing values correspond to the median of each MSHR (i.e. `Region`). BreakpointsThe last position of a region with identical amount of DNA is flagged by 1 otherwise it is 0. Note that during the "Optimization of the number of breakpoints" step, removed breakpoints are flagged by -1. RegionEach position between two breakpoints are labelled the same way with an integer value starting from one. The label is incremented by one when a new breakpoint is found or when moving to the next chromosome. The variable `region` is what we call MSHR. LevelEach position with equal smoothing value is labelled the same way with an integer value starting from one. The label is incremented by one when a new level is found or when moving to the next chromosome. OutliersAwsEach AWS outliers are flagged -1 or 1 otherwise it is 0. OutliersMadEach MAD outliers are flagged -1 (if it is in the α/2 lower tail of the distribution) or 1 (if it is in the α/2 upper tail of the distribution) otherwise it is 0. OutliersTotOutliersAws + OutliersMad. ZoneChrClusters identified after MSHR (i.e. `Region`) clustering by chromosome. ZoneGenClusters identified after HCSR clustering throughout the genome. ZoneGNLStatus of each clone : Gain is coded by 1, Loss by -1 and Normal by 0.
`BkpInfo:` | the data.frame attribute `BkpInfo` which gives the list of breakpoints: PosOrderThe rank position of each clone on the genome. PosBaseThe base position of each clone on the genome. ChromosomeChromosome name.
`SigmaC:` | the data.frame attribute `SigmaC` gives the estimation of the LogRatio standard-deviation for each chromosome: ChromosomeChromosome name. ValueThe estimation is based on the Inter Quartile Range.

### Note

People interested in tools dealing with array CGH analysis can visit our web-page http://bioinfo.curie.fr.

### Author(s)

Philippe HupÃ©, glad@curie.fr.

### References

- HupÃ© et al. (Bioinformatics, 2004) Analysis of array CGH data: from signal ratio to gain and loss of DNA regions.

- Polzehl and Spokoiny (WIAS-Preprint 787, 2002)Local likelihood modelling by adaptive weights smoothing.

- Ben-Yaacov and Eldar (Bioinformatics, 2008)A fast and flexible method for the segmentation of aCGH data.

### See Also

`profileCGH`, `as.profileCGH`, `plotProfile`.

### Examples

    data(snijders)

    ### Creation of "profileCGH" object
    gm13330$Clone <- gm13330$BAC
    profileCGH <- as.profileCGH(gm13330)

    ###########################################################
    ###
    ###  glad function as described in HupÃ© et al. (2004)
    ###
    ###########################################################

    res <- glad(profileCGH, mediancenter=FALSE,
                    smoothfunc="lawsglad", bandwidth=10, round=1.5,
                    model="Gaussian", lkern="Exponential", qlambda=0.999,
                    base=FALSE,
                    lambdabreak=8, lambdacluster=8, lambdaclusterGen=40,
                    type="tricubic", param=c(d=6),
                    alpha=0.001, msize=5,
                    method="centroid", nmax=8,
                    verbose=FALSE)

    ### cytoband data to plot chromosomes
    data(cytoband)

    ### Genomic profile on the whole genome
    plotProfile(res, unit=3, Bkp=TRUE, labels=FALSE, Smoothing="Smoothing",
    main="Breakpoints detection: GLAD analysis", cytoband = cytoband)

    ###Genomic profile for chromosome 1
    plotProfile(res, unit=3, Bkp=TRUE, labels=TRUE, Chromosome=1,
    Smoothing="Smoothing", main="Chromosome 1: GLAD analysis", cytoband = cytoband)

    ### The standard-deviation of LogRatio are:
    res$SigmaC

    ### The list of breakpoints is:
    res$BkpInfo

[Package GLAD version 2.50.0 Index]