// Copyright (C) The Lightning Authors. All rights reserved. // // SPDX-License-Identifier: AGPL-3.0 package lightning import ( "bufio" "bytes" "context" "errors" "flag" "fmt" "io" "io/ioutil" "net/http" _ "net/http/pprof" "os" "sort" "strconv" "strings" "sync" "sync/atomic" "git.arvados.org/arvados.git/sdk/go/arvados" "github.com/arvados/lightning/hgvs" "github.com/kshedden/gonpy" "github.com/sirupsen/logrus" log "github.com/sirupsen/logrus" ) type exportNumpy struct { filter filter } func (cmd *exportNumpy) RunCommand(prog string, args []string, stdin io.Reader, stdout, stderr io.Writer) int { var err error defer func() { if err != nil { fmt.Fprintf(stderr, "%s\n", err) } }() flags := flag.NewFlagSet("", flag.ContinueOnError) flags.SetOutput(stderr) pprof := flags.String("pprof", "", "serve Go profile data at http://`[addr]:port`") runlocal := flags.Bool("local", false, "run on local host (default: run in an arvados container)") projectUUID := flags.String("project", "", "project `UUID` for output data") priority := flags.Int("priority", 500, "container request priority") inputDir := flags.String("input-dir", "./in", "input `directory`") outputDir := flags.String("output-dir", "./out", "output `directory`") annotationsFilename := flags.String("output-annotations", "", "output `file` for tile variant annotations csv") librefsFilename := flags.String("output-onehot2tilevar", "", "when using -one-hot, create csv `file` mapping column# to tag# and variant#") labelsFilename := flags.String("output-labels", "", "output `file` for genome labels csv") regionsFilename := flags.String("regions", "", "only output columns/annotations that intersect regions in specified bed `file`") expandRegions := flags.Int("expand-regions", 0, "expand specified regions by `N` base pairs on each side`") onehot := flags.Bool("one-hot", false, "recode tile variants as one-hot") chunks := flags.Int("chunks", 1, "split output into `N` numpy files") cmd.filter.Flags(flags) err = flags.Parse(args) if err == flag.ErrHelp { err = nil return 0 } else if err != nil { return 2 } if *pprof != "" { go func() { log.Println(http.ListenAndServe(*pprof, nil)) }() } if !*runlocal { runner := arvadosContainerRunner{ Name: "lightning export-numpy", Client: arvados.NewClientFromEnv(), ProjectUUID: *projectUUID, RAM: 500000000000, VCPUs: 96, Priority: *priority, KeepCache: 1, APIAccess: true, } err = runner.TranslatePaths(inputDir, regionsFilename) if err != nil { return 1 } runner.Args = []string{"export-numpy", "-local=true", "-pprof", ":6060", fmt.Sprintf("-one-hot=%v", *onehot), "-input-dir", *inputDir, "-output-dir", "/mnt/output", "-output-annotations", "/mnt/output/annotations.csv", "-output-onehot2tilevar", "/mnt/output/onehot2tilevar.csv", "-output-labels", "/mnt/output/labels.csv", "-regions", *regionsFilename, "-expand-regions", fmt.Sprintf("%d", *expandRegions), "-chunks", fmt.Sprintf("%d", *chunks), } runner.Args = append(runner.Args, cmd.filter.Args()...) var output string output, err = runner.Run() if err != nil { return 1 } fmt.Fprintln(stdout, output+"/matrix.npy") return 0 } tilelib := &tileLibrary{ retainNoCalls: true, retainTileSequences: true, compactGenomes: map[string][]tileVariantID{}, } err = tilelib.LoadDir(context.Background(), *inputDir) if err != nil { return 1 } log.Info("filtering") cmd.filter.Apply(tilelib) log.Info("tidying") tilelib.Tidy() log.Info("building lowqual map") lowqual := lowqual(tilelib) names := cgnames(tilelib) if *labelsFilename != "" { log.Infof("writing labels to %s", *labelsFilename) var f *os.File f, err = os.OpenFile(*labelsFilename, os.O_CREATE|os.O_WRONLY, 0777) if err != nil { return 1 } defer f.Close() outBasename := "matrix.npy" for i, name := range names { _, err = fmt.Fprintf(f, "%d,%q,%q\n", i, trimFilenameForLabel(name), outBasename) if err != nil { err = fmt.Errorf("write %s: %w", *labelsFilename, err) return 1 } } err = f.Close() if err != nil { err = fmt.Errorf("close %s: %w", *labelsFilename, err) return 1 } } log.Info("determining which tiles intersect given regions") dropTiles, err := chooseTiles(tilelib, *regionsFilename, *expandRegions) if err != nil { return 1 } annotation2tvs := map[string]map[hgvs.Variant][]tileLibRef{} if *annotationsFilename != "" { log.Info("writing annotations") var annow io.WriteCloser annow, err = os.OpenFile(*annotationsFilename, os.O_CREATE|os.O_WRONLY, 0666) if err != nil { return 1 } defer annow.Close() var mtx sync.Mutex err = (&annotatecmd{ maxTileSize: 5000, dropTiles: dropTiles, reportAnnotation: func(tag tagID, _ int, variant tileVariantID, refname string, seqname string, pdi hgvs.Variant) { mtx.Lock() defer mtx.Unlock() if annotation2tvs[seqname] == nil { annotation2tvs[seqname] = map[hgvs.Variant][]tileLibRef{} } annotation2tvs[seqname][pdi] = append(annotation2tvs[seqname][pdi], tileLibRef{Tag: tag, Variant: variant}) }, }).exportTileDiffs(annow, tilelib) if err != nil { return 1 } err = annow.Close() if err != nil { return 1 } } var lastErr atomic.Value var wg sync.WaitGroup for seqname, pdivars := range annotation2tvs { seqname, pdivars := seqname, pdivars wg.Add(1) go func() { defer wg.Done() log.Infof("choosing hgvs columns for seq %s", seqname) var pdis []hgvs.Variant for pdi, librefs := range pdivars { // Include this HGVS column if it was // seen in a variant of any // non-dropped tile. for _, libref := range librefs { if int(libref.Tag) >= len(dropTiles) || !dropTiles[libref.Tag] { pdis = append(pdis, pdi) break } } } sort.Slice(pdis, func(i, j int) bool { if cmp := pdis[i].Position - pdis[j].Position; cmp != 0 { return cmp < 0 } else if pdis[i].Ref != pdis[j].Ref { return pdis[i].Ref < pdis[j].Ref } else { return pdis[i].New < pdis[j].New } }) log.Infof("writing column labels for seq %s", seqname) var buf bytes.Buffer for _, pdi := range pdis { fmt.Fprintf(&buf, "%s:g.%s\n", seqname, pdi.String()) } err := ioutil.WriteFile(*outputDir+"/"+seqname+".columns.csv", buf.Bytes(), 0777) if err != nil { lastErr.Store(err) return } log.Infof("building hgvs matrix for seq %s", seqname) data := make([]int8, len(names)*len(pdis)*2) for row, name := range names { cg := tilelib.compactGenomes[name] rowstart := row * len(pdis) * 2 for col, pdi := range pdis { for _, libref := range pdivars[pdi] { if len(cg) <= int(libref.Tag)*2+1 { continue } for phase := 0; phase < 2; phase++ { if cg[int(libref.Tag)*2+phase] == libref.Variant { data[rowstart+col*2+phase] = 1 } } } } } log.Infof("writing hgvs numpy for seq %s", seqname) f, err := os.OpenFile(*outputDir+"/"+seqname+".npy", os.O_CREATE|os.O_WRONLY, 0777) if err != nil { lastErr.Store(err) return } defer f.Close() // gonpy closes our writer and ignores errors. Give it a nopCloser so we can close f properly. npw, err := gonpy.NewWriter(nopCloser{f}) if err != nil { lastErr.Store(err) return } npw.Shape = []int{len(names), len(pdis) * 2} err = npw.WriteInt8(data) if err != nil { lastErr.Store(err) return } err = f.Close() if err != nil { lastErr.Store(err) return } }() } wg.Wait() if e, ok := lastErr.Load().(error); ok { err = e return 1 } chunksize := (len(tilelib.variant) + *chunks - 1) / *chunks for chunk := 0; chunk < *chunks; chunk++ { log.Infof("preparing chunk %d of %d", chunk+1, *chunks) tagstart := chunk * chunksize tagend := tagstart + chunksize if tagend > len(tilelib.variant) { tagend = len(tilelib.variant) } out, rows, cols := cgs2array(tilelib, names, lowqual, dropTiles, tagstart, tagend) var npw *gonpy.NpyWriter var output io.WriteCloser fnm := *outputDir + "/matrix.npy" if *chunks > 1 { fnm = fmt.Sprintf("%s/matrix.%d.npy", *outputDir, chunk) } output, err = os.OpenFile(fnm, os.O_CREATE|os.O_WRONLY, 0777) if err != nil { return 1 } defer output.Close() bufw := bufio.NewWriter(output) npw, err = gonpy.NewWriter(nopCloser{bufw}) if err != nil { return 1 } if *onehot { log.Info("recoding to onehot") recoded, librefs, recodedcols := recodeOnehot(out, cols) out, cols = recoded, recodedcols if *librefsFilename != "" { log.Infof("writing onehot column mapping") err = cmd.writeLibRefs(*librefsFilename, tilelib, librefs) if err != nil { return 1 } } } log.WithFields(logrus.Fields{ "filename": fnm, "rows": rows, "cols": cols, }).Info("writing numpy") npw.Shape = []int{rows, cols} npw.WriteInt16(out) err = bufw.Flush() if err != nil { return 1 } err = output.Close() if err != nil { return 1 } } return 0 } func (*exportNumpy) writeLibRefs(fnm string, tilelib *tileLibrary, librefs []tileLibRef) error { f, err := os.OpenFile(fnm, os.O_CREATE|os.O_WRONLY, 0666) if err != nil { return err } defer f.Close() for i, libref := range librefs { _, err = fmt.Fprintf(f, "%d,%d,%d\n", i, libref.Tag, libref.Variant) if err != nil { return err } } return f.Close() } func cgnames(tilelib *tileLibrary) (cgnames []string) { for name := range tilelib.compactGenomes { cgnames = append(cgnames, name) } sort.Slice(cgnames, func(i, j int) bool { return trimFilenameForLabel(cgnames[i]) < trimFilenameForLabel(cgnames[j]) }) return } func lowqual(tilelib *tileLibrary) (lowqual []map[tileVariantID]bool) { lowqual = make([]map[tileVariantID]bool, len(tilelib.variant)) for tag, variants := range tilelib.variant { lq := lowqual[tag] for varidx, hash := range variants { if len(tilelib.hashSequence(hash)) == 0 { if lq == nil { lq = map[tileVariantID]bool{} lowqual[tag] = lq } lq[tileVariantID(varidx+1)] = true } } } return } func cgs2array(tilelib *tileLibrary, names []string, lowqual []map[tileVariantID]bool, dropTiles []bool, tagstart, tagend int) (data []int16, rows, cols int) { rows = len(tilelib.compactGenomes) for tag := tagstart; tag < tagend; tag++ { if len(dropTiles) <= tag || !dropTiles[tag] { cols += 2 } } data = make([]int16, rows*cols) for row, name := range names { cg := tilelib.compactGenomes[name] outidx := 0 for tag := tagstart; tag < tagend && tag*2+1 < len(cg); tag++ { if len(dropTiles) > tag && dropTiles[tag] { continue } for phase := 0; phase < 2; phase++ { v := cg[tag*2+phase] if v > 0 && lowqual[tag][v] { data[row*cols+outidx] = -1 } else { data[row*cols+outidx] = int16(v) } outidx++ } } } return } func makeMask(regionsFilename string, expandRegions int) (*mask, error) { log.Printf("makeMask: reading %s", regionsFilename) rfile, err := zopen(regionsFilename) if err != nil { return nil, err } defer rfile.Close() regions, err := io.ReadAll(rfile) if err != nil { return nil, err } log.Print("makeMask: building mask") var mask mask for _, line := range bytes.Split(regions, []byte{'\n'}) { if bytes.HasPrefix(line, []byte{'#'}) { continue } fields := bytes.Split(line, []byte{'\t'}) if len(fields) < 3 { continue } refseqname := string(fields[0]) if strings.HasPrefix(refseqname, "chr") { refseqname = refseqname[3:] } start, err1 := strconv.Atoi(string(fields[1])) end, err2 := strconv.Atoi(string(fields[2])) if err1 == nil && err2 == nil { // BED } else { start, err1 = strconv.Atoi(string(fields[3])) end, err2 = strconv.Atoi(string(fields[4])) if err1 == nil && err2 == nil { // GFF/GTF end++ } else { return nil, fmt.Errorf("cannot parse input line as BED or GFF/GTF: %q", line) } } mask.Add(refseqname, start-expandRegions, end+expandRegions) } log.Print("makeMask: mask.Freeze") mask.Freeze() return &mask, nil } func chooseTiles(tilelib *tileLibrary, regionsFilename string, expandRegions int) (drop []bool, err error) { if regionsFilename == "" { return } mask, err := makeMask(regionsFilename, expandRegions) if err != nil { return } tagset := tilelib.taglib.Tags() if len(tagset) == 0 { err = errors.New("cannot choose tiles by region in a library without tags") return } taglen := len(tagset[0]) log.Print("chooseTiles: check ref tiles") // Find position+size of each reference tile, and if it // intersects any of the desired regions, set drop[tag]=false. // // (Note it doesn't quite work to do the more obvious thing -- // start with drop=false and change to true when ref tiles // intersect target regions -- because that would give us // drop=false for tiles that don't appear at all in the // reference.) // // TODO: (optionally?) don't drop tags for which some tile // variants are spanning tiles, i.e., where the reference tile // does not intersect the desired regions, but a spanning tile // from a genome does. drop = make([]bool, len(tilelib.variant)) for i := range drop { drop[i] = true } for refname, refseqs := range tilelib.refseqs { for refseqname, reftiles := range refseqs { if strings.HasPrefix(refseqname, "chr") { refseqname = refseqname[3:] } tileend := 0 for _, libref := range reftiles { if libref.Variant < 1 { err = fmt.Errorf("reference %q seq %q uses variant zero at tag %d", refname, refseqname, libref.Tag) return } seq := tilelib.TileVariantSequence(libref) if len(seq) < taglen { err = fmt.Errorf("reference %q seq %q uses tile %d variant %d with sequence len %d < taglen %d", refname, refseqname, libref.Tag, libref.Variant, len(seq), taglen) return } tilestart := tileend tileend = tilestart + len(seq) - taglen if mask.Check(refseqname, tilestart, tileend) { drop[libref.Tag] = false } } } } log.Print("chooseTiles: done") return } func recodeOnehot(in []int16, incols int) (out []int16, librefs []tileLibRef, outcols int) { rows := len(in) / incols maxvalue := make([]int16, incols) for row := 0; row < rows; row++ { for col := 0; col < incols; col++ { if v := in[row*incols+col]; maxvalue[col] < v { maxvalue[col] = v } } } outcol := make([]int, incols) dropped := 0 for incol, maxv := range maxvalue { outcol[incol] = outcols if maxv == 0 { dropped++ } for v := 1; v <= int(maxv); v++ { librefs = append(librefs, tileLibRef{Tag: tagID(incol), Variant: tileVariantID(v)}) outcols++ } } log.Printf("recodeOnehot: dropped %d input cols with zero maxvalue", dropped) out = make([]int16, rows*outcols) for inidx, row := 0, 0; row < rows; row++ { outrow := out[row*outcols:] for col := 0; col < incols; col++ { if v := in[inidx]; v > 0 { outrow[outcol[col]+int(v)-1] = 1 } inidx++ } } return } type nopCloser struct { io.Writer } func (nopCloser) Close() error { return nil } func trimFilenameForLabel(s string) string { if i := strings.LastIndex(s, "/"); i >= 0 { s = s[i+1:] } s = strings.TrimSuffix(s, ".gz") s = strings.TrimSuffix(s, ".fa") s = strings.TrimSuffix(s, ".fasta") s = strings.TrimSuffix(s, ".1") s = strings.TrimSuffix(s, ".2") s = strings.TrimSuffix(s, ".gz") s = strings.TrimSuffix(s, ".vcf") s = strings.Replace(s, ",", "-", -1) return s }