Description Usage Arguments Details Value Note Author(s) See Also Examples

View source: R/accessory_functions.R

`twosamplecompare`

"resegments" two samples to have the same breakpoints. Both samples' means of the resulting segments are tested for equality using the two-sided Welch two sample t-test. `twosamplecompare`

returns a data frame with the comparisons per segment, it returns the correlation of segments, and a copy number plot with an overlay of (scaled) segment values of both samples and the associated -log10-transformed q-values.

1 2 3 4 5 6 | ```
twosamplecompare(template1, index1 = FALSE, ploidy1 = 2,
cellularity1 = 1, standard1, name1, template2, index2 = FALSE,
ploidy2 = 2, cellularity2 = 1, standard2, name2,
equalsegments = FALSE, altmethod = FALSE, cap = 12, qcap = 12,
bottom = 0, plot = TRUE, trncname = FALSE, legend = TRUE,
chrsubset, onlyautosomes = TRUE, showcorrelation = TRUE)
``` |

`template1` |
Object. Either a data frame as created by |

`index1` |
Integer. If template1 is a QDNAseqobject, this specifies the index of the first sample. Default = FALSE |

`ploidy1` |
Integer. Assume the median of segments of the first sample has this absolute copy number. Default = 2 |

`cellularity1` |
Numeric. Used for rescaling bin and segment values of the first sample. Default = 1 |

`standard1` |
Numeric. Forces ploidy1 to represent this raw value. When omitted, the standard will be calculated from the data. When using parameters obtained from |

`name1` |
Character string. Name of the first sample. Printed on graph |

`template2` |
Object. Either a data frame as created by |

`index2` |
Integer. Specifies the index of the second sample in template2 or, when template2 is omitted, in template1. Default = FALSE |

`ploidy2` |
Integer. Assume the median of segments of the second sample has this absolute copy number. Default = 2 |

`cellularity2` |
Numeric. Used for rescaling bin and segment values of the second sample. Default = 1 |

`standard2` |
Numeric. Forces ploidy2 to represent this raw value. When omitted, the standard will be calculated from the data. When using parameters obtained from |

`name2` |
Character string. Name of the second sample. Printed on graph |

`equalsegments` |
Logical or integer. If TRUE, |

`altmethod` |
Logical or character string. Instead of scaling the sample segments to absolute copies, scale them to standard units. There are two options: "SD" and "MAD". In the first case, the mean of segments is set to 0 and for each segment the distance (in standard deviations or "SD units" from the segment mean to the mean of segments is calculated in standard deviations. In case of "MAD", instead the median of segments, segment median, and median absolute deviation is used. Adjust the y-axis with the cap and bottom arguments for better visualization. Default = FALSE |

`cap` |
Integer. Influences your output copy number graphs. The upper limit of the y-axis is set at this number. When set to "max", it sets the cap to the maximum absolute copynumber value, rounded up. Bins and segments that exceed the cap are represented by a special mark. Recommended use between 8 and 16. Default = 12 |

`qcap` |
Integer. Sets the upper limit of the secondary y-axis. Default = 12 |

`bottom` |
Integer. Similar to cap, but for the lower limit of the y-axis. When set to "min", it sets the bottom to the minimum absolute copynumber value, rounded down. Bins and segments that subceed the bottom are represented by a special mark. Default = 0 |

`plot` |
Logical. Produce a two-sample copy number plot. Default = TRUE |

`trncname` |
Logical. In case of a QDNAseq object, the name of the sample is retrieved from the object and used as title. If set to TRUE, |

`legend` |
Logical. Add the legend to the two-sample plot. Default = TRUE |

`chrsubset` |
Integer vector. Specify the chromosomes you want to plot. It will always take the full range of chromosomes in your subset, so specifying chrsubset = c(4, 8) will give the same plot as chrsubset = 4:8. When using a subset, |

`onlyautosomes` |
Logical or integer. You can fill in an integer to specify how many autosomes your species has. When TRUE, |

`showcorrelation` |
Logical. Add the correlation to the two-sample plot. Default = TRUE |

This function can be used for different types of comparisons. It can be used to compare a tumor sample with a healthy (preferably matched) control. In this case, it may not be necessary to fill in the cellularity, because it will not make a difference for the statistical tests. In this ability the function helps to determine which (if any) segments are significantly different from normal. The other major use is to compare two tumors from potentially the same origin, but that were separated in space or time. You can then assess if changes have occurred, or even whether the two samples are from different clonal origin. In this case it is important to achieve maximum similarity in segments. Now the argument altmethod may come in handy, because it does not require model fitting and optimization. The q-values that are obtained with this function should be interpreted with caution. The two-sample statistical tests will easily reach significance when the sample sizes, in this case bins per segment, are large. By creating equal segment sizes with the argument equalsegments, these biases disappear.

twosampledf - data frame with the newly created segments and the information and comparison of both samples

correlation - Pearson correlation of the segment values of all bins between both samples

subsetcorrelation - same as correlation, but only applying to subset of chromosomes specified by the argument chrsubset

compareplot - ggplot2-graph of both samples with segment values in red (first sample) and blue (second sample). Green bars indicate q-values of the segments, scaled on the secondary axis

The dataframe returned by this function always contains all segments, even when the argument chrsubset is used. However, if there is no useful data for an entire chromosome, it will not constitute a segment and thus be excluded from the data frame, even though the chromsome may be included in the plot. If you want to know the correlation of segments of a subset of chromosomes, you need to have the argument plot = TRUE, because this particular correlation number is calculated in the plot loop. If you do not want to render the plot, you can follow the closing parenthesis of the function call with $subsetcorrelation. If you want to get rid of the green significance bars in the plot, you can set qcap = 0. If you insist on getting rid of the entire secondary axis, save the plot to an object, then run: plotobject + scale_y_continuous(name = "copies", sec.axis = sec_axis(~., labels = NULL), expand = c(0,0))

Jos B. Poell

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 | ```
## simulated data assuming each chromosome comprises 100 bins
s1 <- jitter(c(1, 1, 0.8, 1.2, rep(1, 5), 1.4, rep(1, 13)), amount = 0)
s2 <- jitter(c(1, 1, 1.25, rep(1, 5), 1.5, rep(1, 13)), amount = 0)
n1 <- c(100, 100, 40, 60, rep(100, 5), 100, rep(100, 13))
n2 <- c(rep(100, 22))
bin <- 1:2200
chr <- rep(1:22, each = 100)
start <- rep(0:99*1000000+1, 22)
end <- rep(1:100*1000000, 22)
copynumbers1 <- jitter(rep(s1,n1), amount = 0.05)
copynumbers2 <- jitter(rep(s2,n2), amount = 0.05)
segments1 <- rep(s1, n1)
segments2 <- rep(s2, n2)
template1 <- data.frame(bin = bin, chr = chr, start = start, end = end,
copynumbers = copynumbers1, segments = segments1)
template2 <- data.frame(bin = bin, chr = chr, start = start, end = end,
copynumbers = copynumbers2, segments = segments2)
twosamplecompare(template1 = template1, template2 = template2,
cellularity1 = 0.4, cellularity2 = 0.5)
twosamplecompare(template1 = template1, template2 = template2,
cellularity1 = 0.4, cellularity2 = 0.5, equalsegments = 20)
## using segmented data from a QDNAseq-object
data("copyNumbersSegmented")
## for derivations of the parameters for this fit, see squaremodel
twosamplecompare(copyNumbersSegmented, index1 = 1, cellularity1 = 0.4,
standard1 = 1, index2 = 2, cellularity2 = 0.41, ploidy2 = 2.08,
standard2 = 1)
``` |

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