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Stoye, J.

Publications and source records attributed to Stoye, J..

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Quantifying the Rearrangement Complexity of Pangenomes

The study of evolution between species (phylogenetics) and the study of evolution within a species (population genetics) are highly related, as the same biological mechanisms are fundamental to both fields. Although both have been studied for a long time, their joint study in a unified setting has been prevented by the different time scales they consider and the different data types they employ. A similar discrepancy holds for their whole-genome specializations, comparative genomics and pangenomics. Two active areas in these fields are genome rearrangement studies and graphical pangenomics, respectively. Since the emergence of graphical pangenomics, these have existed as separate fields, despite observations that central data structures representing genomic variants in both fields are highly similar. While there exists a wealth of theoretical results for various rearrangement models in comparative genomics, the application to pangenomic data is hampered by the limitations of rearrangement problem formulations. On the practical side, pangenomes typically contain too many individual genomes for classical problems, such as the often NP-hard parsimony problems, to be solved, or for all-vs-all comparisons using rearrangement distances to be performed. On the theoretical side, some assumptions in the formulation of rearrangement problems, such as the assumption of an underlying tree, are inadequate for many pangenomes. In this work, we propose the Complete Ancestral Reconstruction for Pangenomes (CARP) problem, which overcomes these limitations while retaining intuitive relationships to both classical rearrangement problems and pangenome graphs.

bioinformatics↗

On Deriving Synteny Blocks by Compacting Elements

Genomic rearrangements are major drivers of evolution and genetic disease. However, studying rearrangements requires segmenting the genomes of interest into conserved regions, called synteny blocks, that highlight structural differences between genomes. Synteny blocks are typically defined from annotated genes or derived as a by-product of whole-genome alignments. As these procedures are heuristic and do not explicitly model rearrangements, they can obscure real variation, create false similarities, and affect phylogenetic inference. The importance of synteny block definition has long been recognized, as shown for example by discussions on breakpoint reuse, where different definitions of synteny blocks led to different estimates of rearrangement complexity in mammalian genomes. We present a formal framework for deriving synteny blocks directly from sequence data by partitioning genomic elements into blocks that do not contain breakpoints. A breakpoint is defined between a pair of genomes as an adjacency of shared elements that occurs in one genome but not in the other. Synteny blocks are therefore not allowed to span such boundaries, ensuring that rearrangements are not obscured. The framework is fully agnostic to the type of genomic element and applies to any genome representation expressed as sequences of elements, such as non-overlapping alignments, exact matches (MUMs/MEMs), k-mers, unitigs or minimizers. We formalize two optimization problems: minimizing the total genome length after replacement by synteny blocks (the Minimum-Length Synteny Block Problem) and minimizing the number of distinct blocks (the Minimum-Size Synteny Block Problem). We show that both problems are NP-hard in general. However, when blocks are required to be collinear and to contain a shared element, we provide a linear-time algorithm with respect to the number of input elements that simultaneously minimizes both objectives. The resulting method is simple, efficient, and produces large synteny blocks without obscuring rearrangements.

bioinformatics↗

Panacus: fast and exact pangenome growth and core size estimation

MotivationUsing a single linear reference genome poses a limitation to exploring the full genomic diversity of a species. The release of a draft human pangenome underscores the increasing relevance of pangenomics to overcome these limitations. Pangenomes are commonly represented as graphs, which can represent billions of base pairs of sequence. Presently, there is a lack of scalable software able to perform key tasks on pangenomes, such as quantifying universally shared sequence across genomes (the core genome) and measuring the extent of genomic variability as a function of sample size (pangenome growth). ResultsWe introduce Panacus (pangenome-abacus), a tool designed to rapidly perform these tasks and visualize the results in interactive plots. Panacus can process GFA files, the accepted standard for pangenome graphs, and is able to analyze a human pangenome graph with 110 million nodes in less than one hour. AvailabilityPanacus is implemented in Rust and is published as Open Source software under the MIT license. The source code and documentation are available at https://github.com/marschall-lab/panacus. Panacus can be installed via Bioconda at https://bioconda.github.io/recipes/panacus/README.html. ContactLuca Parmigiani (luca.parmigiani@uni-bielefeld.de), Daniel Doerr (daniel.doerr@hhu.de).

bioinformatics↗

Applying rearrangement distances to enable plasmid epidemiology with pling

Plasmids are a key vector of antibiotic resistance, but the current bioinformatics toolkit is not well suited to tracking them. The rapid structural changes seen in plasmid genomes present considerable challenges to evolutionary and epidemiological analysis. Typical approaches are either low resolution (replicon typing) or use shared k-mer content to define a genetic distance. However this distance can both overestimate plasmid relatedness by ignoring rearrangements, and underestimate by over-penalising gene gain/loss. Therefore a model is needed which captures the key components of how plasmid genomes evolve structurally - through gene/block gain or loss, and rearrangement. A secondary requirement is to prevent promiscuous transposable elements (TEs) leading to over-clustering of unrelated plasmids. We choose the "Double Cut and Join Indel" model, in which plasmids are studied at a coarse level, as a sequence of signed integers (representing genes or aligned blocks), and the distance between two plasmids is the minimum number of rearrangement events or indels needed to transform one into the other. We show how this gives much more meaningful distances between plasmids. We introduce a software workflow pling (https://github.com/iqbal-lab-org/pling), which uses the DCJ-Indel model, to calculate distances between plasmids and then cluster them. In our approach, we combine containment distances and DCJ-Indel distances to build a TE-aware plasmid network. We demonstrate superior performance and interpretability to other plasmid clustering tools on the "Russian Doll" dataset and a hospital transmission dataset. Impact statementStudying plasmid transmission is a necessary component of understanding antibiotic resistance spread, but identifying recently related plasmids is difficult and often requires manual curation. Pling simplifies this by leveraging a combination of containment distances and rearrangement distances to cluster plasmids. The outcome are clusters of recently related plasmids with a clear backbone and relatively large core genomes, in contrast to other tools which sometimes overcluster. Additionally the network constructed by pling provides a framework with which to spot evolutionary events, such as potential fusions of plasmids and spread of transposable elements. Data summarySupplementary information and figures are available as an additional PDF. The tool presented in this paper is available under https://github.com/iqbal-lab-org/pling. Additional computational analysis and scripts are described and provided under https://github.com/babayagaofficial/pling_paper_analyses. The sequence data used can be found under BioProject no. PRJNA246471 in the National Center for Biotechnology Information for the "Russian doll" dataset (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA246471), and under Project no. PRJEB31034 in European Nucleotide Archive for the "Addenbrookes" dataset (https://www.ebi.ac.uk/ena/browser/view/PRJEB30134). All other genome sequences used were sourced from PLSDB (https://ccb-microbe.cs.uni-saarland.de/plsdb/), and lists of accession numbers can be found in the additional analysis github.

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Revisiting pangenome openness with k-mers

Pangenomics is the study of related genomes collectively, usually from the same species or closely related taxa. Originally, pangenomes were defined for bacterial species. After the concept was extended to eukaryotic genomes, two definitions of pangenome evolved in parallel: the gene-based approach, which defines the pangenome as the union of all genes, and the sequence-based approach, which defines the pangenome as the set of all nonredundant genomic sequences. Estimating the total size of the pangenome for a given species has been subject of study since the very first mention of pangenomes. Traditionally, this is performed by predicting the ratio at which new genes are discovered, referred to as the openness of the species. Here, we abstract each genome as a set of items, which is entirely agnostic of the two approaches (gene-based, sequence-based). Genes are a viable option for items, but also other possibilities are feasible, e.g., genome sequence substrings of fixed length k (k-mers). In the present study, we investigate the use of k-mers to estimate the openness as an alternative to genes, and compare the results. An efficient implementation is also provided.

bioinformatics↗

Detecting High Scoring Local Alignments in Pangenome Graphs

MotivationIncreasing amounts of individual genomes sequenced per species motivate the usage of pangenomic approaches. Pangenomes may be represented as graphical structures, e.g. compacted colored de Bruijn graphs, which offer a low memory usage and facilitate reference-free sequence comparisons. While sequence-to-graph mapping to graphical pangenomes has been studied for some time, no local alignment search tool in the vein of BLAST has been proposed yet. ResultsWe present a new heuristic method to find maximum scoring local alignments of a DNA query sequence to a pangenome represented as a compacted colored de Bruijn graph. Our approach additionally allows a comparison of similarity among sequences within the pangenome. We show that local alignment scores follow an exponential-tail distribution similar to BLAST scores, and we discuss how to estimate its parameters to separate local alignments representing sequence homology from spurious findings. An implementation of our method is presented, and its performance and usability are shown. Our approach scales sublinearly in running time and memory usage with respect to the number of genomes under consideration. This is an advantage over classical methods that do not make use of sequence similarity within the pangenome.

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Reconstructing tumor evolutionary histories and clone trees in polynomial-time with SubMARine

Tumors contain multiple subpopulations of genetically distinct cancer cells. Reconstructing their evolutionary history can improve our understanding of how cancers develop and respond to treatment. Subclonal reconstruction methods cluster mutations into groups that co-occur within the same subpopulations, estimate the frequency of cells belonging to each subpopulation, and infer the ancestral relationships among the subpopulations by constructing a clone tree. However, often multiple clone trees are consistent with the data and current methods do not efficiently capture this uncertainty; nor can these methods scale to clone trees with a large number of subclonal populations. Here, we formalize the notion of a partial clone tree that defines a subset of the pairwise ancestral relationships in a clone tree, thereby implicitly representing the set of all clone trees that have these defined pairwise relationships. Also, we introduce a special partial clone tree, the Maximally-Constrained Ancestral Reconstruction (MAR), which summarizes all clone trees fitting the input data equally well. Finally, we extend commonly used clone tree validity conditions to apply to partial clone trees and describe SubMARine, a polynomial-time algorithm producing the subMAR, which approximates the MAR and guarantees that its defined relationships are a subset of those present in the MAR. We also extend SubMARine to work with subclonal copy number aberrations and define equivalence constraints for this purpose. In contrast with other clone tree reconstruction methods, SubMARine runs in time and space that scales polynomially in the number of subclones. We show through extensive simulation and a large lung cancer dataset that the subMAR equals the MAR in > 99.9% of cases where only a single clone tree exists and that it is a perfect match to the MAR in most of the other cases. Notably, SubMARine runs in less than 70 seconds on a single thread with less than one Gb of memory on all datasets presented in this paper, including ones with 50 nodes in a clone tree. The freely-available open-source code implementing SubMARine can be downloaded at https://github.com/morrislab/submarine. Author summaryCancer cells accumulate mutations over time and consist of genetically distinct subpopulations. Their evolutionary history (as represented by tumor phylogenies) can be inferred from bulk cancer genome sequencing data. Current tumor phylogeny reconstruction methods have two main issues: they are slow, and they do not efficiently represent uncertainty in the reconstruction. To address these issues, we developed SubMARine, a fast algorithm that summarizes all valid phylogenies in an intuitive format. SubMARine solved all reconstruction problems in this manuscript in less than 70 seconds, orders of magnitude faster than other methods. These reconstruction problems included those with up to 50 subclones; problems that are too large for other algorithms to even attempt. SubMARine achieves these result because, unlike other algorithms, it performs its reconstruction by identifying an upper-bound on the solution set of trees. In the vast majority of cases, this upper bound is tight: when only a single solution exists, SubMARine converges to it > 99.9% of the time; when multiple solutions exist, our algorithm correctly recovers the uncertain relationships in more than 80% of cases. In addition to solving these two major challenges, we introduce some useful new concepts for and open research problems in the field of tumor phylogeny reconstruction. Specifically, we formalize the concept of a partial clone tree which provides a set of constraints on the solution set of clone trees; and provide a complete set of conditions under which a partial clone tree is valid. These conditions guarantee that all trees in the solution set satisfy the constraints implied by the partial clone tree.

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HASLR: Fast Hybrid Assembly of Long Reads

Third generation sequencing technologies from platforms such as Oxford Nanopore Technologies and Pacific Biosciences have paved the way for building more contiguous assemblies and complete reconstruction of genomes. The larger effective length of the reads generated with these technologies has provided a mean to overcome the challenges of short to mid-range repeats. Currently, accurate long read assemblers are computationally expensive while faster methods are not as accurate. Therefore, there is still an unmet need for tools that are both fast and accurate for reconstructing small and large genomes. Despite the recent advances in third generation sequencing, researchers tend to generate second generation reads for many of the analysis tasks. Here, we present HASLR, a hybrid assembler which uses both second and third generation sequencing reads to efficiently generate accurate genome assemblies. Our experiments show that HASLR is not only the fastest assembler but also the one with the lowest number of misassemblies on all the samples compared to other tested assemblers. Furthermore, the generated assemblies in terms of contiguity and accuracy are on par with the other tools on most of the samples. AvailabilityHASLR is an open source tool available at https://github.com/vpc-ccg/haslr.

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