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Paulsen, J.

Publications and source records attributed to Paulsen, J..

12 recordsLinked to original sources

Epigenetic Profiling of Human Insulinomas Reveals AP-1 Family as Critical Regulators of Beta Cell Maturation

Insulinomas are rare and benign human pancreatic adenomas that overproduce insulin and display increased beta cell mass. We and others have shown that transcriptomic and genomic profiling on insulinomas provides a data mine for identifying targets that can be manipulated to induce human beta cell regeneration. Majority of causative genetic variants in insulinomas involve epigenetic regulatory genes. Yet, specifically how these variants lead to human beta cell expansion and increased function is largely unknown. Here, we performed bulk and single-nucleus epigenomic and transcriptomic profiling to define regulatory alterations in human insulinomas. Bulk ATAC-seq and H3K27Ac ChIP-seq revealed significant enrichment of AP-1 transcription factor binding motifs within beta cell-associated open chromatin/enhancer regions in normal islets, accompanied by robust expression of AP-1 family members; in contrast, insulinomas exhibited marked reductions in both AP-1 motif enrichment and AP-1 expression. Our snRNA-seq and snATAC-seq profiling across four independent insulinomas identified a consistent and previously unrecognized signature defined by suppression of AP-1 transcription factors and widespread loss of chromatin accessibility at AP-1 binding sites, particularly at enhancers governing beta cell identity. Collectively, these results establish AP-1-mediated regulatory programs as critical determinants of beta cell maturation and define their disruption as a signature among human insulinomas.

molecular biology↗

HP1α depletion and TGFβ activation exert antagonistic effects on 3D genome organization

The three-dimensional (3D) organization of the human genome plays a critical role in regulating gene expression and is frequently disrupted in cancer. However, how key factors like Heterochromatin Protein 1 alpha (HP1) and Transforming Growth Factor beta (TGF{beta}) remodel this architecture to drive tumorigenesis remains poorly understood. We investigated the effects of HP1 knockdown and TGF{beta} treatment on higher-order chromatin structure and gene expression in human mammary epithelial cells. Our findings reveal that HP1 depletion and TGF{beta} stimulation exert distinct and opposing effects on genome compartmentalization and sub compartmentalization. HP1 knockdown drives a genome-wide shift of chromatin from transcriptionally inactive B compartments to active A compartments. This is accompanied by a stepwise redistribution of A subcompartments toward the most transcriptionally active state (A3), and the upregulation of oncogenic genes involved in EMT and proliferation. In contrast, TGF{beta} treatment promotes chromatin compaction, increases the proportion of B compartments, and drives a stepwise reduction in active A subcompartments. Our results highlight the differential roles of HP1 and TGF{beta} in shaping the 3D genome and underscore how precise, stepwise architectural changes contribute to malignant transformation. This study provides critical insight into how chromatin architecture acts as a regulatory layer in breast cancer development. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=138 SRC="FIGDIR/small/705282v1_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@11c7475org.highwire.dtl.DTLVardef@121b305org.highwire.dtl.DTLVardef@1704b64org.highwire.dtl.DTLVardef@1659c34_HPS_FORMAT_FIGEXP M_FIG GRAPHICAL ABSTRACT C_FIG

genomics↗

The 3D nuclear position and compartmentalization of genes prime their response to mechano-confinement

Cells continuously receive mechanical inputs from their environment. For example, during migration in narrow spaces, or in solid tumors, cells and their nuclei experience confinement which they sense and respond to. Emerging evidence identifies the genome as a central mediator of these responses, dynamically reorganizing its three-dimensional (3D) structure to regulate both short- and long-term gene expression programs, enabling cellular adaptation to mechanical constraints. However, the mechanisms underlying such responses, especially those linking the 3D genome and transcriptome, are unknown. Here, we utilize controlled cell confinement followed by Hi-C and transcriptome analyses to map and model temporal responses to mechano-confinement in the 3D genome. We identify clusters of genes (termed "TECs") exhibiting coordinated temporal transcriptional responses tied to their differential radial positioning within the nucleus. Additionally, we uncover a genome-wide, partially reversible response, wherein chromosomes reposition under confinement to enhance select genome compartments, aligning with temporal gene regulation patterns. Specifically, one such cluster, TEC7, is linked to 3D genome restructuring and nuclear translocation of NF-{kappa}B, driving cytokine expression and secretion. Our findings reveal how 3D genome reorganization and transcriptional programs can drive mechanoresponses, opening up for new views into the mechanisms of mechanogenomic regulation in health and disease.

genomics↗

Rarity and reproductive biology drive population genomics of obligate symbioses

Modern perspectives on obligate symbioses increasingly recognize their dynamism, complexity, and global importance. Population genomics has been key to this advancement, yet the extension of existing frameworks to reproducibly integrate across multiple genotype-phenotype-environment relationships remains elusive. We generated 962 metagenomes for six lichens, obligate symbioses between fungi and algae, with contrasting traits sampled at a landscape scale. Core neutral population metrics were primarily driven by rarity and reproductive biology, while genes underlying expression regulation were under selection in response to land use and climate across all species. Examined within a comparative framework, these results illustrate that symbiont interactions are a major factor influencing microevolutionary trajectories, especially neutral processes. We present a conceptual framework for population-scale evolution in obligate symbioses that centers on biotic interactions.

evolutionary biology↗

Helitrons are enriched in lichenized fungi with long generation lengths and small distribution sizes

Mobile genetic elements (MGEs) have the potential to drive genome evolution by introducing mutations and causing structural instability and chromosomal rearrangements, particularly under conditions like environmental or genetic stress. In this study, we generated 18 new long-read based reference genomes for lichenized fungi, which form obligate mutualistic symbioses with algae or cyanobacteria. We used the new genomes and 10 publically available genomes to investigate the relationships between species traits (i.e., dominant reproductive mode, distribution size, and generation length) and the abundance and spatial distribution of MGEs using a phylogenetic comparative framework. We found that species with smaller distribution sizes and longer generation lengths had a higher genomic DNA transposon load. Specifically, their genomes were enriched with Rolling Circle transposons, which contradicts previous research that has identified high proportions of retrotransposons in rare species. Disproportionate distributions of MGEs in rare and range-restricted species may disrupt genomic stability, decrease fitness, and be reflective of species experiencing a greater degree of stress. Conversely, greater MGE activity may be an important source of novel genetic diversity in isolated populations with limited gene flow. Further research is needed to understand the potential mechanisms driving MGE proliferation in rare species genomes, and if MGE content is predictive of increased extinction risk.

genomics↗

StripePy: fast and robust characterization of architectural stripes

MotivationArchitectural stripes in Hi-C and related data are crucial for gene regulation, development, and DNA repair. Despite their importance, few tools exist for automatic stripe detection. ResultsWe introduce StripePy, which leverages computational geometry methods to identify and analyze architectural stripes in contact maps from Chromosome Conformation Capture experiments like Hi-C and Micro-C. StripePy outperforms existing tools, as shown through tests on various datasets and a newly developed simulated benchmark, StripeBench, providing a valuable resource for the community. Availability and implementationStripePy is released to the public as an open source, MIT-licensed Python application. StripePy source code is hosted on GitHub at https://github.com/paulsengroup/StripePy and is archived on Zenodo. StripePy can be easily installed from source or PyPI using pip and from Bioconda using conda. Containerized versions of StripePy are regularly published on DockerHub. Contactandrea.raffo@ibv.uio.no Supplementary informationSupplementary data are provided as a separate file.

bioinformatics↗

hictk: blazing fast toolkit to work with .hic and .cool files

MotivationHi-C is gaining prominence as a method for mapping genome organization. With declining sequencing costs and a growing demand for higher-resolution data, efficient tools for processing Hi-C datasets at different resolutions are crucial. Over the past decade, the .hic and Cooler file formats have become the de-facto standard to store interaction matrices produced by Hi-C experiments in binary format. Interoperability issues make it unnecessarily difficult to convert between the two formats and to develop applications that can process each format natively. ResultsWe developed hictk, a toolkit that can transparently operate on .hic and .cool files with excellent performance. The toolkit is written in C++ and consists of a C++ library with Python and R bindings as well as CLI tools to perform common operations directly from the shell, including converting between .hic and .mcool formats. We benchmark the performance of hictk and compare it with other popular tools and libraries. We conclude that hictk significantly outperforms existing tools while providing the flexibility of natively working with both file formats without code duplication. AvailabilityThe hictk library, Python bindings and CLI tools are released under the MIT license as a multi-platform application available at github.com/paulsengroup/hictk. Pre-built binaries for Linux and macOS are available on bioconda. Python bindings for hictk are available on GitHub at github.com/paulsengroup/hictkpy, while R bindings are available on GitHub at github.com/paulsengroup/hictkR. Contactjonas.paulsen@ibv.uio.no Supplementary informationSupplementary data are available at Bioinformatics online.

bioinformatics↗

Multi-level 3D genome organization deteriorates during breast cancer progression

Breast cancer entails intricate alterations in genome organization and expression. However, how three-dimensional (3D) chromatin structure changes in the progression from a normal to a breast cancer malignant state remains unknown. To address this, we conducted an analysis combining Hi-C data with lamina-associated domains (LADs), epigenomic marks, and gene expression in an in vitro model of breast cancer progression. Our results reveal that while the fundamental properties of topologically associating domains (TADs) are overall maintained, significant changes occur in the organization of compartments and subcompartments. These changes are closely correlated with alterations in the expression of oncogenic genes. We also observe a restructuring of TAD-TAD interactions, coinciding with a loss of spatial compartmentalization and radial positioning of the 3D genome. Notably, we identify a previously unrecognized interchromosomal insertion event, wherein a locus on chromosome 8 housing the MYC oncogene is inserted into a highly active subcompartment on chromosome 10. This insertion is accompanied by the formation of de novo enhancer contacts and activation of MYC, illustrating how structural genomic variants can alter the 3D genome to drive oncogenic states. In summary, our findings provide evidence for the loss of genome organization at multiple scales during breast cancer progression revealing novel relationships between genome 3D structure and oncogenic processes.

genomics↗

Eight principal chromatin states functionally segregate the fly genome into developmental and housekeeping roles

Different chromatin forms, or states, represent a fundamental means of controlling gene regulation. Chromatin states have been studied through either the distribution of histone modifications (e.g. 1-5) or more rarely via the occupancy of chromatin proteins 6-8. However, these two approaches disagree on the nature and composition of active chromatin states 2,9 and modelling chromatin via both histone marks and chromatin proteins has been lacking. Here, combining protein and histone mark profiles, we show that chromatin in Drosophila melanogaster is organised into eight principle chromatin states that have consistent forms and constituents across cell types. These states form through the association of the Swi/Snf chromatin remodelling complex, Polycomb Group (PcG)/H3K27me3, HP1a/H3K9me3 or H3K36me3 complexes with either active complexes (RNA Pol/COM-PASS/H3K4me3/NuRF) or repressive marks (histone H1 and nuclear lamin occupancy). Enhancers, core promoters, transcription factor motifs, and gene bodies show distinct chromatin state preferences that separate by developmental and housekeeping/metabolic gene ontology. Within the 3D genome, chromatin states add an additional level of compartmentalisation through self-association of topologically associated domains (TADs) of the same state. Our results suggest that the epigenetic landscape is organised by the binding of chromatin remodellers and repressive complexes, and that through chromatin states the genome is fundamentally segregated into developmental and housekeeping/metabolic roles.

genomics↗

MoDLE: High-performance stochastic modeling of DNA loop extrusion interactions

DNA loop extrusion emerges as a key process establishing genome structure and function. We introduce MoDLE, a computational tool for fast, stochastic modeling of molecular contacts from DNA loop extrusion capable of simulating realistic contact patterns genome wide in a few minutes. MoDLE accurately simulates contact maps in concordance with existing molecular dynamics approaches and with Micro-C data, and does so orders of magnitude faster than existing approaches. MoDLE runs efficiently on machines ranging from laptops to high performance computing clusters, and opens up for exploratory and predictive modeling of 3D genome structure in a wide range of settings.

bioinformatics↗

TAD cliques predict key features of chromatin organization

Processes underlying genome 3D organization and domain formation in the mammalian nucleus are not completely understood. Multiple processes such as transcriptional compartmentalization, DNA loop extrusion and interactions with the nuclear lamina dynamically act on chromatin at multiple levels. Here, we explore long-range interaction patterns between topologically associated domains (TADs) in several cell types. We find that this is connected to many key features of chromatin organization, including open and closed compartments, compaction and loop extrusion processes. We find that domains that form large TAD cliques tend to be repressive across cell types, when comparing gene expression, LINE/SINE repeat content and chromatin subcompartments. Further, TADs in large cliques are larger in genomic size, less dense and depleted of convergent CTCF motifs, in contrast to smaller and denser TADs formed by a loop extrusion processes. Our results shed light on the organizational principles that govern repressive and active domains in the human genome.

genomics↗

Hi-C-constrained physical models of human chromosomes recover functionally-related properties of genome organization

Combining genome-wide structural models with phenomenological data is at the forefront of efforts to understand the organizational principles regulating the human genome. Here, we use chromosome-chromosome contact data as knowledge- based constraints for large-scale three-dimensional models of the human diploid genome. The resulting models remain minimally entangled and acquire several functional features that are observed in vivo and that were never used as input for the model. We find, for instance, that gene-rich, active regions are drawn towards the nuclear center, while gene poor and lamina-associated domains are pushed to the periphery. These and other properties persist upon adding local contact constraints, suggesting their compatibility with non-local constraints for the genome organization. The results show that suitable combinations of data analysis and physical modelling can expose the unexpectedly rich functionally-related properties implicit in chromosome-chromosome contact data. Specific directions are suggested for further developments based on combining experimental data analysis and genomic structural modelling.

biophysics↗