Search bioRxiv⌕ Search

bioRxiv · 10.1101/085399

Evaluation of the accuracy of imputed sequence variants and their utility for causal variant detection in cattle

Abstract

BackgroundThe availability of dense genotypes and whole-genome sequence variants from various sources offers the opportunity to compile large data sets consisting of tens of thousands of animals with genotypes for millions of polymorphic sites that may enhance the power of genomic analyses. The imputation of missing genotypes ensures that all individuals have genotypes for a shared set of variants.\n\nResultsWe evaluated the accuracy of imputation from dense genotypes to whole-genome sequence variants in 249 Fleckvieh and 450 Holstein cattle using Minimac and FImpute. The sequence variants of a subset of the animals were reduced to the variants that were included in the Illumina BovineHD genotyping array and subsequently inferred in silico using either within- or multi-breed reference populations. The accuracy of imputation varied considerably across chromosomes and dropped at regions where the bovine genome contains segmental duplications. Depending on the imputation strategy, the correlation between imputed and true genotypes ranged from 0.898 to 0.952. The accuracy of imputation was higher with Minimac than FImpute particularly for rare alleles. Considering a multi-breed reference population increased the accuracy of imputation, particularly when FImpute was used to infer genotypes. When the sequence variants were imputed using Minimac, the true genotypes were more correlated to predicted allele dosages than best-guess genotypes. The computing costs to impute 23,256,743 sequence variants in 6958 animals were 10-fold higher with Minimac than FImpute. Association studies with imputed sequence variants revealed seven quantitative trait loci (QTL) for milk fat percentage. Two known causal mutations in the DGAT1 and GHR genes were the most significantly associated variants at two QTL on chromosomes 14 and 20 when Minimac was used to infer genotypes.\n\nConclusionsThe population-based imputation of millions of sequence variants in large cohorts provides accurate genotypes and is computationally feasible. Using a reference population that includes individuals from many breeds increases the accuracy of imputation particularly at low-frequency variants. Considering allele dosages rather than best-guess genotypes as explanatory variables is advantageous for association studies with imputed sequence variants.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Pausch, H., MacLeod, I. M., Fries, R., Emmerling, R., Bowman, P. J., Daetwyler, H. D., Goddard, M. E.. 2016-11-03. Evaluation of the accuracy of imputed sequence variants and their utility for causal variant detection in cattle. https://doi.org/10.1101/085399

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Recurrent mitotic error and purifying selection shape chromosomal mosaicism across human tissues

Mosaic chromosomal alterations (mCAs) have been extensively studied in cancer and clonal hematopoiesis but less so in healthy solid tissues. Here, we characterized mCAs in 19 tissues across 25 donors (aged 42-89+ years) by analyzing >200x whole-genome sequencing data generated by the Somatic Mosaicism across Human Tissues (SMaHT) Network. Across 722 tissue samples, we identified 1,008 autosomal mCAs >1 Mb in length and ubiquitous losses of sex chromosomes. Surprisingly, the highest levels of Y chromosome loss in males (5-10% cell fraction in elderly donors) were observed in the lung, adrenal gland, and heart. These tissues also tended to experience more X loss in females, revealing a gradient of mitotic error burden across human tissues. Autosomal aneuploidies displayed distinct tissue-specific patterns indicative of negative selection against most events, except in the adrenal gland, in which low-level aneuploidy was pervasive. In contrast to the adrenal gland, testis rarely harbored aneuploidies but frequently experienced low-level copy-neutral loss of heterozygosity (CN-LOH), suggesting increased mitotic recombination in spermatogonial stem cells. Recurrent X chromosome losses within tissues of female donors exhibited allelic skews explained by a combination of genetic and epigenetic effects. Together, these results show that the chromosomal landscape of aging human tissues is shaped by genetics and recurrent mutations, most but not all of which are held in check by purifying selection.

genomics↗

A Chromosome Scale Genome Assembly for the Australian blue mussel Mytilus planulatus

Mussels belonging to the genus Mytilus are among the world's most ecologically and economically important bivalve molluscs. Although the genus most likely originated in the Northern Hemisphere, the evolutionary history of Southern Hemisphere lineages, including the timing and number of colonisation events leading to the extant Australian populations, remains debated. Limited genomic resources have hindered the reconstruction of these evolutionary histories. In Australia, distinguishing native Mytilus planulatus from invasive Mytilus galloprovincialis is critical for biosecurity and conservation, but previous molecular approaches lacked sufficient resolution to resolve species boundaries and hybridisation patterns. Here, we present the first chromosome-scale reference genome for M. planulatus. The assembly consists of 14 chromosomes and over 51,000 annotated protein-coding genes per haplotype. Comparative analysis revealed extensive structural variation and widespread hemizygosity despite a largely conserved chromosomal organisation relative to M. galloprovincialis. Population analyses using SNPs and gene presence-absence variation in resequenced individuals confirmed previously inferred patterns of population structure and establish this reference genome as a valuable resource for future studies of genome evolution, adaptation and population genomics in Mytilus.

genomics↗

An Isogenic iPSC Model of LMNA p.R482W Recapitulates Adipocyte Dysfunction in Familial Partial Lipodystrophy Type 2

Familial partial lipodystrophy type 2 (FPLD2) is a rare autosomal dominant disorder caused by pathogenic variants in LMNA, most commonly p.R482W, and is characterized by selective loss of subcutaneous adipose tissue, severe insulin resistance, and dyslipidemia. The cellular mechanisms underlying adipocyte dysfunction in FPLD2 remain incompletely understood, in part because existing in vitro models rely on patient-derived, non-isogenic cell lines or require non- physiologic transgene overexpression to achieve adipogenic differentiation. Here, we describe a CRISPR/Cas9-edited isogenic iPSC model of FPLD2 in which the heterozygous LMNA c.1444C>T (p.R482W) variant was introduced into a healthy donor-derived iPSC line. Using a transgene-free mesenchymal progenitor differentiation protocol, we generated LMNAp.R482W and isogenic control adipocytes and characterized their transcriptomic and functional phenotypes. Bulk RNA- sequencing demonstrated strong concordance in adipogenesis gene expression between genotypes during differentiation (Pearson {rho} = 0.968), yet LMNAp.R482W adipocytes exhibited significant downregulation of key adipogenic, lipid-handling, and extracellular matrix genes compared to isogenic controls. Functionally, LMNAp.R482W adipocytes showed markedly impaired glucose uptake under both basal and insulin-stimulated conditions, consistent with severe insulin resistance. Seahorse extracellular flux analysis revealed a globally quiescent metabolic state with significantly reduced oxygen consumption rate (126.5 vs. 261.5 pmol/min, p < 0.0001) and extracellular acidification rate (27.6 vs. 46.7 mpH/min, p < 0.0001). To validate these findings in human tissue, gene set enrichment analysis results from the iPSC model were compared to single-nucleus RNA-sequencing data from subcutaneous white adipose tissue biopsies of individuals with FPLD2 carrying the LMNA p.R482W variant. Across 37 mutually significant pathways, 59.5% were concordant between the iPSC model and human adipose tissue, including the pathways most central to FPLD2 pathophysiology: adipogenesis, fatty acid metabolism, oxidative phosphorylation, and inflammatory response. These findings establish this isogenic iPSC system as a faithful cellular model of FPLD2 adipocyte biology and a platform for functional characterization of LMNA variants of uncertain significance.

genomics↗