Updated Cyrius to call all star alleles (up to *139) (#3)

Removed the --knownFunction and --includeNewStar options.

README.md

@@ -1,19 +1,18 @@
 # Cyrius: WGS-based CYP2D6 genotyper
-Cyrius is a tool to genotype CYP2D6 from a whole-genome sequencing (WGS) BAM file. Cyrius uses a novel method to solve the problems caused by the high sequence similarity with the pseudogene paralog CYP2D7 and thus is able to detect all star alleles, particularly those that contain structural variants, accurately. Please refer to our [preprint](https://www.biorxiv.org/content/10.1101/2020.05.05.077966v1) for details about the method.   
+Cyrius is a tool to genotype CYP2D6 from a whole-genome sequencing (WGS) BAM file. Cyrius uses a novel method to solve the problems caused by the high sequence similarity with the pseudogene paralog CYP2D7 and thus is able to detect all star alleles, particularly those that contain structural variants, accurately. Please refer to our [preprint](https://www.biorxiv.org/content/10.1101/2020.05.05.077966v2) for details about the method.   
 
 ## Running the program
 
 This Python3 program can be run as follows:
 ```bash
-star_caller.py --manifest MANIFEST_FILE \
-              --genome [19/37/38] \
-              --prefix OUTPUT_FILE_PREFIX \
-              --outDir OUTPUT_DIRECTORY \
-              --threads NUMBER_THREADS
+python3 star_caller.py --manifest MANIFEST_FILE \
+                       --genome [19/37/38] \
+                       --prefix OUTPUT_FILE_PREFIX \
+                       --outDir OUTPUT_DIRECTORY \
+                       --threads NUMBER_THREADS
 ```
 The manifest is a text file in which each line should list the absolute path to an input BAM/CRAM file.
-For CRAM input, it’s suggested to provide the path to the reference fasta file with `--reference` in the command.  
-Additionally, there is an option `--knownFunction` to call only star alleles with known functions, as well as an option `--includeNewStar` to call all star alleles including the newly added, uncurated ones (\*115-\*139) in PharmVar.
+For CRAM input, it’s suggested to provide the path to the reference fasta file with `--reference` in the command.    
 
 ## Interpreting the output  
 

caller/call_cn.py → caller/call_variants.py

@@ -33,7 +33,15 @@
     process_raw_call_gc,
     process_raw_call_denovo,
 )
-from depth_calling.haplotype import get_haplotypes_from_bam, extract_hap
+from depth_calling.haplotype import (
+    get_haplotypes_from_bam,
+    get_haplotypes_from_bam_single_region,
+    extract_hap,
+)
+from depth_calling.snp_count import (
+    get_supporting_reads,
+    get_supporting_reads_single_region,
+)
 
 
 INTRON1_BP_APPROX = 42130500
@@ -93,6 +101,11 @@
     "g.42129042T>C",
     "g.42129174C>A",
     "g.42129180A>T",
+    "g.42127526C>T",
+    "g.42128325A>G",
+    "g.42126877G>A",
+    "g.42127973T>C",
+    "g.42127556T>C",
 ]
 
 
@@ -197,7 +210,7 @@ def good_read(read):
     return read.is_secondary == 0 and read.is_supplementary == 0
 
 
-def get_allele_counts_42128936(bamfile_handle, genome):
+def get_allele_counts_var42128936(bamfile_handle, genome):
     """
     Search for the inserstions at 42128936 defining
     *30/*40/*58 in read sequences
@@ -223,6 +236,23 @@ def get_allele_counts_42128936(bamfile_handle, genome):
     return (ref_read, long_ins_read, short_ins_read)
 
 
+def update_var42128936(
+    var_list, var_alt, var_ref, ref_read, long_ins_read, short_ins_read
+):
+    """
+    Update variant read counts for g42128936.
+    """
+    if "g.42128936-42128937insGGGGCGAAAGGGGCGAAA" in var_list:
+        long_ins_index = var_list.index("g.42128936-42128937insGGGGCGAAAGGGGCGAAA")
+        var_alt[long_ins_index] = long_ins_read
+        var_ref[long_ins_index] = short_ins_read + ref_read
+    if "g.42128936-42128937insGGGGCGAAA" in var_list:
+        short_ins_index = var_list.index("g.42128936-42128937insGGGGCGAAA")
+        var_alt[short_ins_index] = short_ins_read
+        var_ref[short_ins_index] = long_ins_read + ref_read
+    return var_alt, var_ref
+
+
 def call_exon9gc(d6_count, d7_count, full_length_cn):
     """
     Call exon 9 conversion
@@ -257,28 +287,82 @@ def call_exon9gc(d6_count, d7_count, full_length_cn):
     return None
 
 
-def call_var42126938(bamfile, cnvtag, site42126938, base_db, target_positions):
+def call_var42126938(bamfile, full_length_cn, base_db):
     """
-    Call variant g.42126938C>T (gene conversion variant in homology region) 
+    Call variant g.42126938C>T (gene conversion variant in homology region)
     based on read depth and phased haplotypes
     """
-    dcn = {"star5": 3, "cn2": 4}
-    assert cnvtag in dcn
-    full_length_cn = dcn[cnvtag]
-    d6_cn = call_cn_snp(full_length_cn, [site42126938[0]], [site42126938[1]], 0.8)[0]
     var_called = []
     # Whether g.42126938C>T is on the same haplotype as g.42126611C>G
     G_haplotype = False
+    snp_d6, snp_d7 = get_supporting_reads(
+        bamfile, base_db.dsnp1, base_db.dsnp2, base_db.nchr, base_db.dindex
+    )
+    d6_d7_base_count = [snp_d6[-1], snp_d7[-1]]
+    d6_cn = call_cn_snp(
+        full_length_cn, [d6_d7_base_count[0]], [d6_d7_base_count[1]], 0.8
+    )[0]
     if d6_cn is not None and d6_cn < full_length_cn - 2:
-        haplotype_per_read = get_haplotypes_from_bam(bamfile, base_db, target_positions)
+        haplotype_per_read = get_haplotypes_from_bam(
+            bamfile, base_db, range(len(base_db.dsnp1))
+        )
         recombinant_read_count = extract_hap(haplotype_per_read, [0, 2])
         if "12" in recombinant_read_count and sum(recombinant_read_count["12"]) > 1:
             G_hap_count = extract_hap(haplotype_per_read, [1, 2])
             for _ in range(full_length_cn - 2 - d6_cn):
                 var_called.append("g.42126938C>T")
             if "12" in G_hap_count and sum(G_hap_count["12"]) > 1:
                 G_haplotype = True
-    return var_called, G_haplotype
+    return d6_d7_base_count, var_called, G_haplotype
+
+
+def call_var42127526_var42127556(bamfile, cnvtag, base_db):
+    """
+    Call variant g.42127526C>T (gene conversion variant in homology region)
+    based on read depth and phased haplotypes
+    """
+    var_called = []
+    var_ref, var_alt, var_ref_forward, var_ref_reverse = get_supporting_reads_single_region(
+        bamfile, base_db.dsnp1, base_db.nchr, base_db.dindex
+    )
+    var7526_count = [var_ref[0], var_alt[0]]
+    var7556_count = [var_ref[1], var_alt[1]]
+    if cnvtag in CNVTAG_LOOKUP_TABLE:
+        d6_cn = CNVTAG_LOOKUP_TABLE[cnvtag].exon9_to_intron1
+        var7526_cn = call_cn_snp(d6_cn, [var7526_count[1]], [var7526_count[0]])[0]
+        var7556_cn = call_cn_snp(d6_cn, [var7556_count[1]], [var7556_count[0]])[0]
+        haplotype_per_read = get_haplotypes_from_bam_single_region(
+            bamfile, base_db, range(len(base_db.dsnp1))
+        )
+        recombinant_read_count = extract_hap(haplotype_per_read, [0, 1, 2])
+        if "211" in recombinant_read_count and sum(recombinant_read_count["211"]) > 1:
+            for _ in range(var7526_cn):
+                var_called.append("g.42127526C>T")
+        elif "221" in recombinant_read_count and sum(recombinant_read_count["221"]) > 1:
+            for _ in range(min(var7526_cn, var7556_cn)):
+                var_called.append("g.42127526C>T")
+                var_called.append("g.42127556T>C")
+    return var7526_count, var7556_count, var_called
+
+
+def call_var42127803hap(bamfile, cnvtag, base_db):
+    """
+    Call haplotype with regard to g.42127803C>T and g.42127941G>A
+    """
+    diff_haplotype = False
+    if cnvtag == "cn2":
+        haplotype_per_read = get_haplotypes_from_bam_single_region(
+            bamfile, base_db, range(len(base_db.dsnp1))
+        )
+        recombinant_read_count = extract_hap(haplotype_per_read, [0, 1])
+        if (
+            "12" in recombinant_read_count
+            and sum(recombinant_read_count["12"]) > 1
+            and "21" in recombinant_read_count
+            and sum(recombinant_read_count["21"]) > 1
+        ):
+            diff_haplotype = True
+    return diff_haplotype
 
 
 def get_called_variants(var_list, cn_prob_processed, starting_index=0):

caller/cnv_hybrid.py

@@ -115,6 +115,11 @@ def get_cnvtag(total_cn, rawv, cn_call_per_site, exon9gc_call_stringent, spacer_
                 and exon9gc_call_stringent <= exon9_intron4_sites_consensus
             ):
                 exon9region_sites_consensus = exon9gc_call_stringent
+            elif (
+                exon9region_sites_consensus > exon9gc_call_stringent
+                and exon9gc_call_stringent >= exon9_intron4_sites_consensus
+            ):
+                exon9region_sites_consensus = exon9gc_call_stringent
     else:
         exon9region_sites = [
             a

caller/construct_star_table.py

@@ -22,8 +22,8 @@
 
 
 # Exon 9 gene conversion
-EXON9GC_ALLELES = ["*36", "*4N", "*57", "*83"]
-EXON9GC_PAIR_ALLELES = {"*36": "*10", "*4N": "*4A"}
+EXON9GC_ALLELES = ["*36", "*4.013", "*57", "*83"]
+EXON9GC_PAIR_ALLELES = {"*36": "*10", "*4.013": "*4"}
 
 
 def make_hap_dic(variant_list, star_set, hap_dic):
@@ -51,7 +51,7 @@ def get_hap_table(hap_table):
         for line in f:
             at = line.strip().split()
             star_id = at[0]
-            variant_list = sorted(at[1:-2])
+            variant_list = sorted(at[1:-1])
             var_list_joined = "_".join(variant_list)
             dhap.setdefault(var_list_joined, star_id)
             dstar.setdefault(star_id, var_list_joined)