See edit below Using R, I would like to filter a matrix (of gene expression data) and keep only the rows (genes/probes) that have values with high variance. For example, I'd like to only keep the rows that have values in the bottom and top percentiles (e.g. below 20% and above 80%). I want to limit my study to only genes under high variance for downstream analyses. Are there common ways for gene filtering in R?
My matrix has 18 samples (columns) and 47000 probes (rows) with values that are log2 transformed and normalized. I know the quantile()
function can identify the 20% and 80% cutoffs within each sample column. I can't figure out how to find these values for the entire matrix, and then subset the original matrix to remove all the "non-varying" rows.
Example matrix with a mean of 5.97, thus the last three rows should be removed because they contain values between the 20% and 80% cutoffs:
> m
sample1 sample2 sample3 sample4 sample5 sample6
ILMN_1762337 7.86 5.05 4.89 5.74 6.78 6.41
ILMN_2055271 5.72 4.29 4.64 5.00 6.30 8.02
ILMN_1736007 3.82 6.48 6.06 7.13 8.20 4.06
ILMN_2383229 6.34 4.34 6.12 6.83 4.82 5.57
ILMN_1806310 6.15 6.37 5.54 5.22 4.59 6.28
ILMN_1653355 7.01 4.73 6.62 6.27 4.77 6.12
ILMN_1705025 6.09 6.68 6.80 6.85 8.35 4.15
ILMN_1814316 5.77 5.17 5.94 6.51 7.12 7.20
ILMN_1814317 5.97 5.97 5.97 5.97 5.97 5.97
ILMN_1814318 5.97 5.97 5.97 5.97 5.97 5.97
ILMN_1814319 5.97 5.97 5.97 5.97 5.97 5.97
I'd appreciate any suggestions, or functions that I should look into. Thanks!
EDIT
Sorry, I was not very clear in the OP. (1) I'd like to know the 20% and 80% cutoff values for the entire matrix (not just for each individual sample). (2) Then, if any row contains a value in the upper or lower percentiles, R will keep these rows. If a row contains values (for all samples) that fall near the mean, these rows are thrown out.
Ok, assuming you have a matrix (so I am assuming that your ID column is actually rownames) then this is very simple to do.
# First find the desired quantile breaks for the entire matrix
qt <- quantile( m , probs = c(0.2,0.8) )
# 20% 80%
#5.17 6.62
# Next get a logical vector of the rows that have any values outside these breaks
rows <- apply( m , 1 , function(x) any( x < qt[1] | x > qt[2] ) )
# Subset on this vector
m[ rows , ]
# sample1 sample2 sample3 sample4 sample5 sample6
#ILMN_1762337 7.86 5.05 4.89 5.74 6.78 6.41
#ILMN_2055271 5.72 4.29 4.64 5.00 6.30 8.02
#ILMN_1736007 3.82 6.48 6.06 7.13 8.20 4.06
#ILMN_2383229 6.34 4.34 6.12 6.83 4.82 5.57
#ILMN_1806310 6.15 6.37 5.54 5.22 4.59 6.28
#ILMN_1653355 7.01 4.73 6.62 6.27 4.77 6.12
#ILMN_1705025 6.09 6.68 6.80 6.85 8.35 4.15
#ILMN_1814316 5.77 5.17 5.94 6.51 7.12 7.20
The any( x < qt[1] | x > qt[2] )
part of the apply
function (which is designed to apply a function across the margins of a matrix) returns TRUE
if any value in that row is outside the 20% and 80% quantiles of your sample matrix. By definition, if no value is outside these bounds it returns FALSE
indicating we will drop that row in the next line.
The Biocondcutor genefilter package provides common filters relevant to microarray analysis. A typical filter based on row-wise variability would be
m = matrix(rnorm(47000 * 6), 47000)
varFilter(m)
The package landing page references vignettes illustrating basic operation and providing diagnostic guidance for use of filtering.
A basic principle in the analysis of microarrays is that values in a row are comparable, but not values between rows. This is because the probes associated with each row have distinct characteristics that introduce row-specific bias -- a value in the first row could reasonably indicate more, less or equal gene expression compared to a value for the same sample in a second row. This means that @Todd's desire to normalize based on between-row comparison (largest and smallest values in the entire matrix) is not recommended. Instead, varFilter calculates a measure of variability of each row (row inter-quartile range) and selects a fraction (the var.cutoff argument) with most variability.
A quick peak at the definition of varFilter
shows that in general this is no more tricky than, for some measure of row-wise variability var.func
and a (single) quantile var.cutoff
vars <- apply(m, 1, var.func)
m[vars > quantile(vars, var.cutoff), ]
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