This project contains code for simultaneously detecting family inheritance patterns, identity-by-descent, crossovers, and inherited deletions in nuclear families, as published in

Paskov K, Chrisman B, Stockham N, Washington PY, Dunlap K, Jung JY, Wall DP. Identifying crossovers and shared genetic material in whole genome sequencing data from families. Genome Research. 2023 Oct 1;33(10):1747-56.

This code starts with VCF files (with accompanying .tbi files), split by chromosome. It produces

• family inheritance patterns across the genome (.BED)
• crossovers (.JSON, .BED)
• inherited deletions (.JSON, .BED)

The code adds to a directory structure created by the https://github.com/kpaskov/VCFtoNPZ project.

[data_dir]
- genotypes
- family_genotype_counts
- sequencing_error_rates
- phase
- - info.json
- - crossovers.json
- - gene_conversions.json
- - inherited_deletions.json
- - sibpairs.json
- - inheritance_patterns
- - - [family_1].bed
- - - [family_2].bed
...

The info.json file contains metadata including the reference assembly (GRch37 or GRch38), vcf directory, ped file, sequencing error parameter files, and any flags provided to the phasing algorithm.

The crossovers.json and crossovers.bed files contains all called crossovers in the cohort in .json and .bed format respectively.

The gene_conversions.json and gene_conversions.bed files contains possible gene-conversion events in the cohort in .json and .bed format respectively. Gene conversions are more difficult to detect than crossovers due to their small size. We have not validated the gene conversions called by our algorithm, so they should be examined carefully before use.

sibpairs.json contains all sibling-pairs in the dataset as well as genome-wide autosomal IBD values and chromosome-level IBD values.

The inherited_deletions.json and inherited_deletions.bed files contains all called inherited deletions in the cohort in .json and .bed format respectively.

The [family].bed files contain inheritance patterns for each family, across all chromosomes in .bed format.

using https://github.com/kpaskov/VCFtoNPZ.

using https://github.com/kpaskov/FamilySeqError.

If using whole-genome sequencing data, follow the instructions for estimating sequencing error rates in both low-complexity and high-complexity regions. A good source of low-complexity regions is the supplementary materials file from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4271055/.

The algorithm detects crossovers and inherited deletions. Run using

python phase/phase_chromosome.py [ped_file] [data_dir] [high_complexity_sequencing_error_profile] [low_complexity_sequencing_error_profile]

The script has options

• --detect_inherited_deletions models inherited deletions while phasing.
• --detect_upd models uniparental disomy while phasing. We detect only heterodisomy because isodisomy is essentially indistinguishable from a denovo deletion. (This option is still under development.)
• --chrom [chrom] phases a single chromosome. If this option is not used, all autosomal chromosomes are phased.
• --family_size [family_size] only families of size [family_size] are phased.
• --family [family] only [family] is phased.
• --batch_size and --batch_num options can be used to parallelize when running on a large cohort.
• --phase_name [name] phase data will be written to directory [data_dir]/phase_[name]/inheritance_patterns. If this option is not used, phase data will be written to directory [data_dir]/phase/inheritance_patterns.

The example below phases all autosomal chromosomes for all families of size 4 in the ssc.hg38 dataset.

python phase/phase_chromosome.py ../DATA/ssc.hg38/ssc.ped HCR LCR --detect_inherited_deletions --family_size 4 

You can use as many sequencing error profiles as you'd like. This comes in handy when phasing the X chromosome, since the PAR and non-PAR regions have different error profiles. This is an example of phasing the X chromosome using four different error profiles.

python phase/phase_chromosome.py ../DATA/ssc.hg38/ssc.ped ../DATA/ssc.hg38 X_PAR_HCR X_PAR_LCR X_nonPAR_HCR X_nonPAR_LCR --chrom X --batch_size 3 --batch_num $SLURM_ARRAY_TASK_ID --detect_inherited_deletions --family_size 4 --phase_name X

If no --chrom option is given, phase_chromosome.py will phase all autosomal chromosomes. The X chromosome must be phased explicitly using the --chrom option.

This script pulls genome-wide autosomal IBD for sibling pairs as well as chromosome-level IBD. It marks siblings who appear to be identical twins or based on their IBD sharing, and identifies siblings with unusual IBD as outliers.

python phase/pull_sibpair_ibd.py [data_dir]

The --phase_name [name] flag indicates that phase data from the directory [data_dir]/phase_[name]/inheritance_patterns will be analyzed.

This script produces [data_dir]/phase/sibpairs.json (or [data_dir]/phase_[name]/sibpairs.json if the --phase_name flag is used) which contains an entry for every sibling pair in the dataset with the following fields:

• family the family ID for the sibling pair
• sibling1 the ID for the first sibling in the pair (the first sibling's ID is alphabetically first)
• sibling2 the ID for the second sibling in the pair (the second sibling's ID is alphabetically last)
• maternal_ibd the genome-wide autosomal maternal IBD between the siblings
• maternal_unknown_fraction the fraction of the autosomal genome where maternal IBD could not be determined
• maternal_ibd_chroms a list of chromosome-level maternal IBD between the siblings in order from chr1-chr22, ending with chrX
• maternal_unknown_fraction_chroms a list of the fraction of each chromosome where maternal IBD could not be determined in order from chr1-chr22, ending with chrX
• paternal_ibd the genome-wide autosomal paternal IBD between the siblings
• paternal_unknown_fraction the fraction of the autosomal genome where paternal IBD could not be determined
• paternal_ibd_chroms a list of chromosome-level paternal IBD between the siblings in order from chr1-chr22, ending with chrX
• paternal_unknown_fraction_chroms a list of the fraction of each chromosome where paternal IBD could not be determined in order from chr1-chr22, ending with chrX
• matxpat_ibd the genome-wide autosomal IBD2 (simultaneous maternal and paternal IBD) between the siblings
• matxpat_unknown_fraction the fraction of the autosomal genome where IBD2 could not be determined
• matxpat_ibd_chroms a list of chromosome-level IBD2 between the siblings in order from chr1-chr22, ending with chrX
• matxpat_unknown_fraction_chroms a list of the fraction of each chromosome where IBD2 could not be determined in order from chr1-chr22, ending with chrX
• is_identical siblings are marked as identical if both maternal and paternal genome-wide autosomal IBD is greater than 0.8.
• is_ibd_outlier siblings are marked as ibd outliers using gaussian kernel density outlier detection

The qc/Visualize-Sibpair-IBD.ipynb jupyter notebook can be used to visualize IBD in the dataset.

This script pulls crossovers for the cohort. It identifies individuals with too many or too few crossovers as outliers.

python phase/pull_crossovers.py [data_dir]

The script has options

• --phase_name [name] flag indicates that phase data from the directory [data_dir]/phase_[name]/inheritance_patterns will be analyzed.
• --hts_loss_regions [loss_region_index1] [loss_region_index2] should be used if multiple sequencing error profiles are used to represent hard-to-sequence regions. For example, the X_chromosome phasing command shown in the example above has two sequencing error profiles corresponding to hard-to-sequence regions (index 1 and 3).

The script produces [data_dir]/phase/crossovers.json and [data_dir]/phase/gene_conversions.json which contain crossover and gene_conversion events respectively. Each event has fields:

• family the family ID where the event was detected
• child the child id the event was detected within
• chrom the chrom where the event ocurred
• start_pos and end_pos the window where the event occured
• is_mat and is_pat indicating whether the event occured during maternal or paternal meiosis
• is_complex true if the event is a complex event (composed of multiple closely spaced events)
• is_hts true if the event occurs in a region marked error-prone for this family

The qc/Visualize-Crossovers.ipynb jupyter notebook can be used to visualize crossovers in the dataset.

This script pulls inherited deletions for the cohort.

python phase/pull_deletions.py [data_dir]

The script has options

• --phase_name [name] flag indicates that phase data from the directory [data_dir]/phase_[name]/inheritance_patterns will be analyzed.

The script produces [data_dir]/phase/deletions.json. Each deletion has fields:

• family the family ID where the deletion was detected
• chrom the chrom where the deletion ocurred
• start_pos and end_pos the window where the deletion occured
• is_mat and is_pat indicating whether the deletion occured during maternal or paternal meiosis
• is_hts true if the deletion occurs in a region marked error-prone for this family
• parent the parent with the deletion
• trans a list of children the deletion was transmitted to
• notrans a list of children the deletion was not transmitted to

The qc/Visualize-Deletions.ipynb jupyter notebook can be used to visualize deletions in the dataset.