Population stratification illustration using slicing of remote VCF with Bioconductor

We will acquire a modest collection of SNP calls from chr17, and project the genotype configurations via principal components to expose population substructure.

Create references to the EBI VCF repository

Bioconductor’s ldblock package includes utilities for working with collections of VCF, typically decomposed by chromosome.

In [21]:
inplist = rownames(installed.packages())
if (!("snpStats" %in% inplist)) install("snpStats")
if (!("ldblock" %in% inplist)) install("vjcitn/ldblock")
if (!("terravar" %in% inplist)) install("vjcitn/terravar")
In [22]:

Read a slice of VCF corresponding to a region of chr17

EBI has updated 1000 genomes calls. We will interrogate a tabix-indexed VCF for chr17 via HTTP. An object that organizes multiple chromosomes is created using stack1kg from the ldblock package.

In [23]:
st = stack1kg("17")

We use a ScanVcfParam to define a slice of genome.

In [24]:
sp = ScanVcfParam(geno="GT", 
    which=GRanges("17", IRanges(32e6,33.75e6)))
myread = readVcfStack(st, param=sp)

readVcfStack produces a CollapsedVCF instance in memory. See the documentation for the VariantAnnotation package for details on this data structure.

In [25]:
class: CollapsedVCF 
dim: 49194 2548 
  GRanges with 5 metadata columns: paramRangeID, REF, ALT, QUAL, FILTER
  DataFrame with 12 columns: AF, AC, NS, AN, EAS_AF, EUR_AF, AFR_AF, AMR_AF,...
             Number Type    Description                                        
   AF        A      Float   Estimated allele frequency in the range (0,1)      
   AC        A      Integer Total number of alternate alleles in called geno...
   NS        1      Integer Number of samples with data                        
   AN        1      Integer Total number of alleles in called genotypes        
   EAS_AF    A      Float   Allele frequency in the EAS populations calculat...
   EUR_AF    A      Float   Allele frequency in the EUR populations calculat...
   AFR_AF    A      Float   Allele frequency in the AFR populations calculat...
   AMR_AF    A      Float   Allele frequency in the AMR populations calculat...
   SAS_AF    A      Float   Allele frequency in the SAS populations calculat...
   VT        .      String  indicates what type of variant the line represents 
   EX_TARGET 0      Flag    indicates whether a variant is within the exon p...
   DP        1      Integer Approximate read depth; some reads may have been...
  SimpleList of length 1: GT
      Number Type   Description    
   GT 1      String Phased Genotype

Transform the VCF content to rare allele counts

We use tools in David Clayton’s snpStats package to achieve a compact representation of (possibly uncertain) genotype calls. This enables us to filter to SNP with MAF exceeding a given threshold, and, later, to compute a PCA.

In [26]:
mymat = genotypeToSnpMatrix(myread)
## non-single nucleotide variations are set to NA
cs = col.summary(mymat[[1]])
sum(cs[,"MAF"]>.1, na.rm=TRUE)
non-single nucleotide variations are set to NA

Build a matrix of B-allele counts

We'll use SNP with MAF exceeding 10%, and build the sample x SNP matrix.

In [27]:
kpsnp = which(cs[,"MAF"]>.1)
cmat = matrix(0, nr=nrow(mymat[[1]]), nc=length(kpsnp))
for (i in seq_len(length(kpsnp))) {
  cmat[,i] = as(mymat[[1]][,kpsnp[i]], "numeric")
rownames(cmat) = rownames(mymat[[1]])

Compute principal components and plot

In [28]:
pp = prcomp(cmat)
In [29]:
  1. 2548
  2. 2548
In [30]:

rownames(geog_1kg) = geog_1kg[,1]
newdf = data.frame(pp$x[,1:4], pop=geog_1kg[rownames(pp$x), "Population"],
    superpop = geog_1kg[rownames(pp$x), "superpop"])
In [31]:
ggplot(newdf, aes(x=PC1, y=PC2, colour=superpop)) + geom_point()