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Szatkiewicz, J.

Publications and source records attributed to Szatkiewicz, J..

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3D-genome impacts of TAD-boundary-disrupting ultra-rare structural variants identified in schizophrenia

Schizophrenia is a complex and multifactorial brain disorder that impacts ~1% of the U.S. population. The genetic architecture of schizophrenia includes 10 large rare structural variants (SVs) ~10 exonic variants, and over 250 common variants. Previously we combined whole genome sequencing (WGS) with brain-specific epigenomic data and found an excess of ultrarare SVs in schizophrenia localizing to the boundaries of topologically associated domains (TADs). Alterations in TAD boundaries (TAD-b) may lead to disarrangement of enhancer-promoter contacts and dysregulation of gene expression and have been associated with several human diseases. Our study sought to functionally characterize two of these TAD-b SVs using chromatin conformation capture (e.g., Hi-C) and RNA sequencing, to unveil mechanistic insight into the effects of these variants on 3D-genome organization and gene expression. To achieve this, we genetically engineered these two TAD-b SVs in isogenic induced pluripotent stem cells (iPSCs) using CRISPR/Cas9. iPSCs were differentiated into neuronal precursor cells, excitatory glutamatergic neurons, and medium spiny neurons. We found that these TAD-boundary-disrupting SVs could reshape the 3D-genome and transcriptional networks in a cell-type-specific manner.

genomics↗

Title: Leveraging Base Pair Mammalian Constraint to Understand Genetic Variation and Human Disease

Although thousands of genomic regions have been associated with heritable human diseases, attempts to elucidate biological mechanisms are impeded by a general inability to discern which genomic positions are functionally important. Evolutionary constraint is a powerful predictor of function that is agnostic to cell type or disease mechanism. Here, single base phyloP scores from the whole genome alignment of 240 placental mammals identified 3.5% of the human genome as significantly constrained, and likely functional. We compared these scores to large-scale genome annotation, genome-wide association studies (GWAS), copy number variation, clinical genetics findings, and cancer data sets. Evolutionarily constrained positions are enriched for variants explaining common disease heritability (more than any other functional annotation). Our results improve variant annotation but also highlight that the regulatory landscape of the human genome still needs to be further explored and linked to disease.

genomics↗

Association Tests Using Copy Number Profile Curves (CONCUR) Enhances Power in Rare Copy Number Variant Analysis

Copy number variants (CNVs) are the gain or loss of DNA segments in the genome that can vary in dosage and length. CNVs comprise a large proportion of variation in human genomes and impact health conditions. To detect rare CNV association, kernel-based methods have been shown to be a powerful tool because their flexibility in modeling the aggregate CNV effects, their ability to capture effects from different CNV features, and their ability to accommodate effect heterogeneity. To perform a kernel association test, a CNV locus needs to be defined so that locus-specific effects can be retained during aggregation. However, CNV loci are arbitrarily defined and different locus definitions can lead to different performance depending on the underlying effect patterns. In this work, we develop a new kernel-based test called CONCUR (i.e., Copy Number profile Curve-based association test) that is free from a definition of locus and evaluates CNV-phenotype association by comparing individuals copy number profiles across the genomic regions. CONCUR is built on the proposed concepts of \"copy number profile curves\" to describe the CNV profile of an individual, and the \"common area under the curve (cAUC) kernel\" to model the multi-feature CNV effects. Compared to existing methods, CONCUR captures the effects of CNV dosage and length, accounts for the continuous nature of copy number values, and accommodates between- and within-locus etiological heterogeneities without the need to define artificial CNV loci as required in current kernel methods. In a variety of simulation settings, CONCUR shows comparable and improved power over existing approaches. Real data analyses suggest that CONCUR is well powered to detect CNV effects in gene pathways associated with phenotypes using data from the Swedish Schizophrenia Study and the Taiwan Biobank.\n\nAuthor summaryCopy number variants comprise a large proportion of variation in human genomes. Large rare CNVs, especially those disrupting genes or changing the dosages of genes, can carry relatively strong risks for neurodevelopmental and neuropsychiatric disorders. Kernel-based association methods have been developed for the analysis of rare CNVs and shown to be a valuable tool. Kernel methods model the collective effect of rare CNVs using flexible kernel functions that capture the characteristics of CNVs and measure CNV similarity of individual pairs. Typically kernels are created by summarizing similarity within an artificially defined \"CNV locus\" and then collapsing across all loci. In this work, we propose a new kernel-based test, CONCUR, that is based on the CNV location information contained in standard processing of the variants and removes the need for any arbitrarily defined CNV loci. CONCUR quantifies similarity between individual pairs as the common area under their copy number profile curves and is designed to detect CNV dosage, length and dosage-length interaction effects. In simulation studies and real data analysis, we demonstrate the ability of CONCUR test to detect CNV effects under diverse CNV architectures with power and robustness over existing methods.

bioinformatics↗

Characterization of single gene copy number variants in schizophrenia

Genetic studies of schizophrenia (SCZ) have now implicated numerous genomic loci that contribute to risk including several copy number variants (CNV) of large effect and hundreds of associated loci of small effect. However, in only a few cases has a specific gene been clearly identified. Rare CNV that affect only a single gene offer a potential avenue to discovering specific SCZ risk genes. Here, we use CNV generated from exome-sequencing of 4,913 SCZ cases and 6,188 controls in a homogenous Swedish cohort to assess the contribution of single-gene deletions and duplications to SCZ risk. As previously seen, we found an excess of rare deletions (p = 0.0004) and duplications (p = 0.0006) in SCZ cases compared to controls. When limiting to only single-gene CNV we identified nominally significant excess of deletions (p = 0.04) and duplications (p = 0.03). In an effort to increase the number of single-gene CNV, we reduced strict filtering criteria but required support from two independent CNV calling methods to create an expanded set that showed a significant burden of deletions in 11 out of 22 gene sets previously implicated in SCZ and in the combined set of genes across those sets (p = 0.008). Finally, for the significantly enriched set of voltage-gated calcium channels, we performed an extensive validation of all deletions generated from exome-sequencing as well as any deletion with evidence from previously analyzed genotyping arrays. In total, 4 exonic, single-gene deletions validated in cases and none in controls (p = 0.039), of which all were identified by exome-sequencing. Broadly, these results point to the potential contribution of single-gene CNV to SCZ and the added value of a deeper dive into CNV calls from exome-sequencing.

genetics↗

Common Variant Associations with Fragile X Syndrome

Fragile X syndrome is a common cause of intellectual disability. It is usually caused by a de novo mutation which often occur on multiple haplotypes and should not be detectible using genome-wide association (GWA). We conducted GWA 89 male FXS cases and 266 male controls, and detected multiple genome-wide significant signals near FMR1 (odds ratio=8.10, P=2.5x10-10). These findings withstood robust attempts at falsification. Fine-mapping did not serve to narrow the interval (minimum P=1.13xl0-14), and functional genomic integration (including 5C data we generated for this region) did not provide a mechanistic hypothesis. Controls carrying a risk haplotype had significantly longer and more variable FMR1 CGG repeats than controls with the protective haplotype (P=4.75x10-5) which may predispose toward increases in CGG number to the pre-mutation range over many generations. This is a salutary reminder of the complexity of even \"simple\" monogenetic disorders.

genetics↗