Genome::Model::Tools::Music::MutationRelation(3pm)

--bam-list
    Provide a file containing sample names and normal/tumor BAM locations for each. Use the tab- delimited format [sample_name normal_bam tumor_bam] per line. This tool only needs sample_name, so all other columns can be skipped. The sample_name must be the same as the tumor sample names used in the MAF file (16th column, with the header Tumor_Sample_Barcode).

EOS
    ); }

sub _doc_authors {
  return <<EOS
 Nathan D. Dees, Ph.D.
 Qunyuan Zhang, Ph.D. EOS }

sub execute {
  # Parse input arguments
  my $self = shift;
  my $bam_list = $self->bam_list;
  my $maf_file = $self->maf_file;
  my $output_file = $self->output_file;
  my $gene_list = $self->gene_list;
  my $permutations = $self->permutations;
  my @all_sample_names; # names of all the samples, no matter if it's mutated or not
  my @genes_to_test; # the genes which will be tested for relationships
  my $skip_non_coding = $self->skip_non_coding;
  my $skip_silent = $self->skip_silent;

  # Parse out the names of the samples which should match the names in the MAF file
  my $sampleFh = IO::File->new( $bam_list ) or die "Couldn't open $bam_list. $!\n";
  my $line_count = 0;
  while( my $line = $sampleFh->getline )
  {
      $line_count++;
      next if ( $line =~ m/^#/ );
      chomp( $line );
      my ( $sample ) = split( /\t/, $line );
      if ($sample) {
          push( @all_sample_names, $sample );
      } else {
          warn("could not parse sample name from line $line_count of --bam_list");
      }
  }
  $sampleFh->close;

  # If user-specified, parse out the names of the genes that we are limiting our tests to
  if( defined $gene_list )
  {
      my $geneFh = IO::File->new( $gene_list ) or die "Couldn't open $gene_list. $!\n";
      while( my $line = $geneFh->getline )
      {
          next if ( $line =~ m/^#/ );
          chomp( $line );
          my ( $gene ) = split( /\t/, $line );
          push( @genes_to_test, $gene );
      }
      $geneFh->close;
  }

  # Create sample-gene matrix
  my $matrix_file = $self->create_sample_gene_matrix($maf_file, \@all_sample_names, \@genes_to_test, $skip_non_coding, $skip_silent);

  # Perform mutation-relation test using R
  my $R_cmd = "R --slave --args < " . __FILE__ . ".R $matrix_file $permutations $output_file";
  print "$R_cmd\n";
  WIFEXITED(system $R_cmd) or croak "Couldn't run: $R_cmd ($?)";

  return(1);
}

sub create_sample_gene_matrix {
    my $self = shift;
    my ( $maf_file, $all_sample_names_ref, $genes_to_test_ref, $skip_non_coding, $skip_silent ) = @_;
    my @all_sample_names = @{$all_sample_names_ref};
    my @genes_to_test = @{$genes_to_test_ref};

    # Create hash of mutations from the MAF file
    my %mutations;
    my %all_genes;

    # Parse the MAF file
    my $mafFh = IO::File->new( $maf_file ) or die "Couldn't open $maf_file. $!\n";
    while( my $line = $mafFh->getline )
    {
        next if( $line =~ m/^(#|Hugo_Symbol)/ );
        chomp $line;
        my @cols = split( /\t/, $line );
        my ( $gene, $mutation_class, $sample ) = ( $cols[0], $cols[8], $cols[15] );

        # If the mutation classification is odd, quit with error
        if( $mutation_class !~ m/^(Missense_Mutation|Nonsense_Mutation|Nonstop_Mutation|Splice_Site|Translation_Start_Site|Frame_Shift_Del|Frame_Shift_Ins|In_Frame_Del|In_Frame_Ins|Silent|Intron|RNA|3'Flank|3'UTR|5'Flank|5'UTR|IGR|Targeted_Region|De_novo_Start_InFrame|De_novo_Start_OutOfFrame)$/ )
        {
            print STDERR "Unrecognized Variant_Classification \"$mutation_class\" in MAF file for gene $gene\n";
            print STDERR "Please use TCGA MAF Specification v2.3.\n";
            return undef;
        }

        # If user wants, skip Silent mutations, or those in Introns, RNA, UTRs, Flanks, IGRs, or the ubiquitous Targeted_Region
        if(( $skip_non_coding && $mutation_class =~ m/^(Intron|RNA|3'Flank|3'UTR|5'Flank|5'UTR|IGR|Targeted_Region)$/ ) ||
            ( $skip_silent && $mutation_class =~ m/^Silent$/ ))
        {
            print "Skipping $mutation_class mutation in gene $gene.\n";
            next;
        }

        $all_genes{$gene}++;
        $mutations{$sample}{$gene}++;
    }
    $mafFh->close;

    # If the user specified a gene list, then check for genes that are not in the MAF
    if( scalar( @genes_to_test ) > 0 )
    {
        for( my $i = 0; $i < scalar( @genes_to_test ); ++$i )
        {
            unless( defined $all_genes{$genes_to_test[$i]} )
            {
                print "Skipping ", $genes_to_test[$i], " from specified gene-list since it was not found in the MAF file\n";
                splice( @genes_to_test, $i, 1 );
            }
        }
    }
    else
    {
        @genes_to_test = sort keys %all_genes;
    }

    # Write the input matrix to a file for use by the R code
    my $matrix_file;
    unless( $matrix_file = $self->mutation_matrix_file ) {
        $matrix_file = Genome::Sys->create_temp_file_path();
    }

    my $matrix_fh = new IO::File $matrix_file,"w";
    # Print input matrix file header
    my $header = join("\t","Sample",@genes_to_test);
    $matrix_fh->print("$header\n");

    # Print mutation relation input matrix
    for my $sample (sort @all_sample_names) {
        $matrix_fh->print($sample);
        for my $gene (@genes_to_test) {
            if (exists $mutations{$sample}{$gene}) {
                $matrix_fh->print("\t1");
            }
            else {
                $matrix_fh->print("\t0");
            }
        }
        $matrix_fh->print("\n");
    }

    return $matrix_file;
}

1;