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Nilsson, M.

Publications and source records attributed to Nilsson, M..

At least 19 recordsLinked to original sources

Improving Data Quality, Model Transparency and Performance in Lung Histopathology with Explainable AI

Convolutional neural networks (CNNs) have shown strong capabilities for image analysis. However, deploying these models in medical settings is complicated by their limited transparency. Over recent years, many approaches have been developed to overcome the so-called "black box" problem of deep neural networks. Here, we show how such explainable AI (XAI) approaches can be applied to not only improve transparency but also training data quality and model performance, with classification of lung damage in histopathology images as use case. First, we conducted a thorough exploratory data analysis and visually compared the compressed multi-dimensional representations of the histology images from the last CNN layers with labels given by pathologists to reveal flaws in the training data. Second, we used Gradient-based Class Activation Mapping (Grad-CAM) as well as SHapley Additive exPlanations (SHAP) values to identify image regions that significantly contributed to the model decisions. To overcome identified shortcomings, we then finetuned additional top layers of the CNNs and introduced model architectures with attention which improved model performance. In summary, we developed a practical workflow that uses interpretability analyses to examine model perception, assess label consistency, and guide model refinement on the example of lung histopathology scoring, demonstrating how XAI approaches can increase both transparency and model performance. The code for this paper is shared at https://github.com/Aitslab/Histology_XAI.git.

pathology↗

Highly resolved tumor architecture via matched spatial and nucleus transcriptomics from a single tissue section

Spatial transcriptomics often relies on reference-based deconvolution to infer cell types in a tissue context; however, public single-cell datasets can miss patient-specific biology. Here we introduce SIMPlex, a method that generates matched spatial and single-nucleus gene-expression profiles from the same 5 um FFPE section. We demonstrate context-matched profiles across mouse brain, breast cancer and prostate cancer tissues, resolving fine-grained cell-states with distinct spatial signatures. By extracting both spatial and nuclear layers, SIMPlex maximises the information recovered from a single tissue section, an advantage for scarce archival and clinical specimens.

molecular biology↗

Protein semi-synthesis enables real-time optical tracking of intracellular conformational changes during sodium channel inactivation

Dynamic conformational changes in intracellular domains fundamentally affect the function and pharmacology of many membrane proteins. For example, sodium influx through the cardiac voltage-gated sodium channel (NaV1.5) is rapidly terminated through conformational changes that result in pore closure, a transition known as inactivation. Inactivation involves NaV1.5 intracellular regions, particularly the DIII-DIV linker containing the IFM particle (Isoleucine-Phenylalanine-Methionine) and its dysfunction is a major cause of cardiac arrythmias. However, the conformational changes involved in inactivation and their modulation by auxiliary proteins and clinically used drugs remain incompletely characterized, partly because live-cell, site-specific labeling of intracellular regions with small fluorescent dyes remains challenging. Here, we combine live-cell protein semi-synthesis with voltage-clamp fluorometry (VCF), to track intracellular conformational dynamics of the cardiac sodium channel NaV1.5 and monitor their voltage dependence and kinetics in real time. We identify intracellular conformational changes involved in both fast and steady-state inactivation of NaV1.5 and show that both lidocaine and auxiliary proteins affect the kinetics of conformational changes of the DIII-DIV linker. Our work establishes the combination of protein semi-synthesis and voltage-clamp fluorometry as a powerful approach to dissect intracellular conformational changes in membrane proteins.

biophysics↗

Spatial microRNA profiling at single-cell resolution by in situ barcoded extension

The spatial organization of post-transcriptional regulation is a fundamental yet difficult to access layer of tissue biology. MicroRNAs (miRNAs) are small RNAs with a key role in post-transcriptional regulation, but their short length has excluded them from spatial profiling technologies, leaving them largely unexplored in spatial transcriptomics. Here, we introduce miR-Space, a method that converts individual miRNAs into extended, uniquely barcoded molecules directly in tissue, enabling their spatial detection by in situ sequencing. Across 30 mouse and human brain sections, miR-Space enabled highly multiplexed miRNA profiling at single-molecule and single-cell resolution, joint analysis with mRNA, and implementation on the automated Xenium platform. miR-Space resolved major anatomical regions and cell populations from spatial miRNA expression, identified reproducible cell-associated miRNA signatures, and uncovered previously unknown spatial and cellular distributions of multiple miRNAs. Together, these capabilities establish miR-Space as a framework for integrating miRNAs into spatial transcriptomics, enabling spatial miRNomics at anatomical and single-cell resolution.

genomics↗

Multi-scale modeling of human tissues from spatial transcriptomics with TERRA

Spatial transcriptomics maps gene expression at cellular resolution, revealing how cells organize into multicellular niches. Yet computational analyses remain dataset-specific, without a transferable representation of tissue organization that generalizes across datasets, tasks and tissues or predicts how tissues behave under perturbation. We present TERRA, a foundation model pretrained on 112 million human cells profiled by spatial transcriptomics. From a single pretrained backbone, TERRA yields embeddings at the scale of cells, the genes they express and the neighborhoods in which they reside, and supports spatial in silico perturbation, all applied zero-shot to unseen tissues. At the cell level, in newly generated spatial data for developing pancreas, TERRA identified an islet-associated capillary state which we posit represents a developmental precursor of the mature islet microvasculature. At the gene level, in untreated kidney sections, in silico knockout of immune-checkpoint targets predicted a gene program of immune-checkpoint-blockade-associated nephrotoxicity, which we validated in treatment-exposed tissue and recovered in blood. At the neighborhood level, TERRA mapped macrophages across tissues to identify recurring cross-organ niches, which we term archetypes, including a tumor-boundary niche associated with poor prognosis in kidney cancer. Together, TERRA captures the spatial and multicellular logic of human tissue and predicts, in silico, its response to perturbation, providing a multi-scale framework for tissue biology, therapeutic development and clinical application.

genomics↗

Cophenetic Spatial Topology Embedding reveals multiscale tissue architecture in spatial omics

The spatial organization of tissues emerges from cell interactions across multiple scales, yet current spatial omics analysis tools often emphasize local neighborhoods and may not summarize broader tissue architecture. Here we introduce Cophenetic Spatial Topology Embedding (COSTE), a computational framework that embeds directed nearest-neighbor distance profiles into a hierarchical metric space without requiring the user to define a spatial radius or neighborhood cutoff. COSTE can be applied to cell-level and single-transcript inputs without requiring cell segmentation. It constructs directed distance profiles between cell populations and uses hierarchical clustering to quantify tissue topology. This yields a Spatial Separation Score (SSS), a sample-normalized score from 0 to 1 that summarizes relative spatial separation within an analyzed tissue. We apply COSTE to spatial transcriptomics datasets of pulmonary fibrosis and triple-negative breast cancer (TNBC), where it delineates tissue structures, nominates spatially defined cell states, and highlights disease-or treatment-associated architectural patterns that are not readily captured by local neighborhood-based analyses. Our approach provides an interpretable framework for exploring tissue architecture and cell-cell spatial relationships in spatial omics data.

bioinformatics↗

Targeted sequencing of mutations via RNA-templated gap filling of oligonucleotides for single-cell RNA-seq

We describe a method for targeted sequencing of expressed mutations in single cells. The approach exploits the reverse transcriptase and nick translation activities of the Bst DNA polymerase (Full Length) from Bacillus stearothermophilus (BstFL) to perform an RNA-templated gap filling and ligation between flanking probes. Integrated with probe-based single cell RNAseq workflows, it enables simultaneous whole transcriptomic and multiplexed mutation profiling from the same cell, overcoming key limitations of mutation detection in existing scRNA-seq approaches.

molecular biology↗

Sympathoadrenal Lineage Plasticity Drives Intratumoral Heterogeneity in Paraganglioma, Neuroblastoma and Composite Tumors Following KIF1Bb-NF1 Loss

Neuroblastoma (NB) and paraganglioma (PPGL) arise from the sympathoadrenal lineage, yet their developmental relationship and heterogeneity remain unclear. Single-cell and spatial transcriptomics revealed shared, spatially organized developmental states, including populations characteristic of the other tumor type, chromaffin-like cells in NB and neuroblast-like cells in PPGL and hybrids co-expressing adjacent states. Deconvolution of an independent PPGL cohort associated metastatic disease most strongly with chromaffin hybrid states, alongside connecting progenitor-like, cycling neuroblast, and early chromaffin identities, whereas non-metastatic tumors were enriched for differentiated late chromaffin identities. In mice, combined loss of the candidate 1p36 tumor suppressor KIF1B{beta} and NF1 recapitulates these developmental architectures by prolonging developmental plasticity, reactivating embryonic neurogenic programs, and driving chromaffin-to-neuroblast transitions generating pheochromocytoma, neuroblastoma and composite tumors. Mouse tumors contained discrete spatial domains of developmental and neoplastic states that mirrored those in human PPGL. Together, our findings establish chromaffin-neuroblast plasticity as a mechanism of sympathoadrenal tumor heterogeneity and identify hybrid developmental states as a potential indicator of plasticity associated with metastatic PPGL.

cancer biology↗

Heterogeneous pro-inflammatory response to BRAFV600E-induced thyroid tumor development

BackgroundThe tumor immune microenvironment likely plays a central role in progression of thyroid cancer. As for most other solid tumors, it is unknown if immune dysregulation contributes to earlier, subclinical stages of thyroid tumor development, or whether thyroid tumor heterogeneity might involve differential expression of pro-inflammatory mediators. MethodsThe time course of tumor-associated inflammation was studied in Tg-CreERT2;Braf CA/+ mice representing a model of BRAFV600E-driven papillary thyroid carcinoma (PTC). Tumor growth was estimated by histological examination and magnetic resonance imaging. Cytokine expression was monitored by quantitative RT-PCR, RNAScope and Western blot analyses. ResultsBased on spontaneous BrafCA activation due to leaky Cre activity in a minority of targeted cells tumors developed within a preserved thyroid tissue architecture to multifocal papillary thyroid carcinoma (PTC) over a period of 12 months. Tumorigenesis was accompanied by a gradually increased mRNA and protein expression of interleukin-1beta (IL-1{beta}), interleukin-6 and tumor necrosis factor-alpha (TNF-) starting already before Braf mutant cells commenced neoplastic growth. RNAScope revealed that both follicular cells and stromal cells expressed Il1b whereas Il6 and Tnfa transcripts were mostly confined to neoplastic epithelia. Early cytokine expression was associated with oncogene-induced senescence, whereas during tumor development (3-6 months) and in advanced tumor stages (at 12 months) the cytokine expression pattern differed among glands and tumor foci of the same gland accompanied by a highly variable locoregional lymphocytic infiltration. Oral treatment of mutant mice for 1 month with PLX4720, a vemurafenib prodrug, partially reduced cytokine expression along with inhibited tumor growth and redifferentiation of thyroid function. The magnitude of reduced cytokine expression differed much between glands and among mice of both sexes. ConclusionsThese findings indicate that oncogenic BRAFV600E targeted to the thyroid both stimulates endogenous production of IL-1{beta}, IL-6 and TNF- and recruits inflammatory cells to foci of early tumor development. PTCs of different clonal origin are distinguished by differential expression of pro-inflammatory cytokines. The anti-inflammatory effect of mutant Braf kinase inhibition varies presumably related to heterogeneous tumor development, which evolves from stochastic BrafCA activation suggesting there are clonally different probabilities of acquiring drug resistance among Braf mutant thyroid follicular cells.

cancer biology↗

Degradation of mucin O-glycans by a human gut symbiont requires a complex enzyme repertoire and promotes colonization

Secreted mucins are the major component of the mucus layer that protects intestinal epithelial surfaces by blocking excessive interactions with the microbiota. Mucins are complex glycoproteins decorated with over 100 different O-glycans. Some bacteria can utilize mucins and excessive degradation has been associated with disruption of the mucus barrier and inflammation. Despite the importance of mucins, a detailed enzymatic pathway by which gut bacteria degrade colonic mucin O-glycans and the impact of this process on gut colonization are unknown. Here, we identified >100 genes that are expressed by the symbiont Bacteroides thetaiotaomicron during growth on different O-glycan substrates, revealing effects of glycan structure on gene expression. The characterization of 33 glycoside hydrolase enzymes revealed the pathway for colonic O-glycan degradation by this bacterium. In vivo competition experiments show that multiple exo-acting enzymes targeting mucin capping structures are central to gut colonization and may provide targets to inhibit bacterial mucin degradation.

microbiology↗

Resolving thyroid lineage cell trajectories merging into a dual endocrine gland in mammals

The thyroid has a remarkable evolution, first appearing in invertebrate chordates as an integral exocrine constituent of the pharyngeal endostyle that is transformed into an endocrine gland during metamorphosis in basal vertebrates. In mammals, the thyroid acquires a second endocrine cell type, calcitonin-producing C-cells, which for long were inferred a neural crest origin, shuttled to the embryonic thyroid by the ultimobranchial bodies. However, recent lineage tracing experiments firmly establish these neuroendocrine cells also derive from foregut endoderm. Key questions remaining unanswered are how thyroid primordia independently develop and, unlike in all non-mammalian vertebrates, merge into a dual endocrine organ. Here, by leveraging a single-cell transcriptome atlas derived from mouse pharyngeal endoderm and its subsequent cell fates, we characterize the global gene expression profile of thyroid- and ultimobranchial-derived progenitor cells and identify comprehensive gene regulatory networks of lineage-specific transcription factors and novel target genes predicted to differentially regulate cell proliferation, plasticity and differentiation during development. Spatiotemporal analyses reveal C-cell precursors are triggered to undergo epithelial-mesenchymal transition (EMT) and cell-autonomously down-regulate collagen IV and degrade laminin that delineates the ultimobranchial body epithelium. However, the EMT program is not fully deployed until both cell lineages are mixed and propagate conjointly thus forming the typical thyroid histoarchitecture of follicles and parafollicular C-cells, every follicle/C-cell unit being enveloped by a renewed basement membrane. Mixed-type thyroid carcinoma recapitulates a synchronous lineage growth pattern but only the neuroendocrine tumor cells are able to escape the compound follicle boundaries and become invasive adopting a C-cell precursor-like migratory phenotype.

developmental biology↗

Tissue architecture and immune niches govern ctDNA release in colorectal cancer

Circulating tumor DNA (ctDNA) is central to liquid biopsy-based cancer detection, yet its release into the bloodstream varies widely and remains poorly understood. To define the tissue-level determinants of ctDNA shedding in colorectal cancer (CRC), we integrated tumor-informed plasma sequencing with detailed histopathology, immunophenotyping, spatial transcriptomics, and in situ mutation detection in resectable stages (I-III). ctDNA detectability increased with tumor burden, and high ctDNA shedders exhibited a distinct architectural and microenvironmental phenotype characterized by expanded necrotic pseudolumina, frequent epithelial barrier disruption, and dense myeloid infiltration. Spatial profiling revealed stress-associated malignant programs and a myeloid-rich immune-luminal niche. In situ sequencing confirmed plasma-detected mutations within pseudoluminal debris, identifying these structures as focal reservoirs of shed DNA. These findings provide a mechanistic framework linking tissue architecture, immune remodelling, and spatially organized cell death to ctDNA release with implications for refining liquid biopsy applications.

cancer biology↗

The role of Akkermansia muciniphila sulfatases in colonic mucinutilisation

Akkermansia muciniphila, an obligate mucin degrader, is a major member of the human colonic microbiota and has been associated positive health outcomes. Mucins are complex glycoproteins that contain heavily sulfated O-glycans and form the protective colonic mucus layer. Bacterial carbohydrate sulfatases are required to metabolise these heavily sulfated mucin glycans and excessive bacterial foraging has been associated with several diseases. Sulfatases have been linked with inflammatory bowel disease, making these microbiota enzymes potential drug targets. A. muciniphila expresses carbohydrate sulfatases that can act on colonic mucins yet their roles in its metabolism remain opaque. Our data reveal that A. muciniphila requires glycopeptides/protein forms of colonic mucin for metabolism and its sulfatases have unique adaptations compared to Bacteroides species. Localisation studies reveal that desulfation of N-acetyl-D-glucosamine, but not D-galactose, is exclusively periplasmic. A cell surface sulfatase has a novel carbohydrate binding module that binds to colonic mucin. This paints a contrasting picture of sulfated mucin metabolism by Akkermansia muciniphila versus Bacteroides species. These data will be important for understanding the contexts for Akkermansia muciniphilas positive health correlations.

biochemistry↗

Cell atlas of the developing human meninges reveals a dura origin of meningioma

The vertebrate central nervous system is enveloped by the meninges, consisting of the pia, arachnoid, and dura layers. The arachnoid is hypothesised to give rise to the most common primary intracranial tumours, meningiomas. However, molecular evidence supporting this hypothesis is lacking. There are no effective medical therapies to treat meningiomas that are resistant to local interventions, encumbered by our limited understanding of their cellular origin. To address this limitation in our understanding of meningioma biology, we generated a comprehensive reference single cell and spatial transcriptomic atlas of human fetal meninges at post-conceptional weeks 5-13. We found that the meningeal layers develop concurrently, and identified an inner CDH1-positive dura cell layer expressing tight junction genes consistent with barrier function. We show that transcriptionally, meningioma cells resemble dura-lineage cells, and that common meningioma driver genes were expressed preferentially in the dura lineage. Our findings suggest that meningiomas originate from dura lineage cells. HIGHLIGHTSO_LIscRNA-seq and spatial transcriptomics reveals architecture of human fetal meninges development C_LIO_LIMeningeal layers are formed concurrently, by a gradual refinement of cell states C_LIO_LIA CDH1+ inner dura sublayer expresses tight junction genes like the arachnoid barrier C_LIO_LIMeningioma tumours likely originate from inner dura lineage cells, not arachnoid C_LI

developmental biology↗

In Situ Inference of Copy Number Variations in Image-Based Spatial Transcriptomics

Copy number variations (CNVs) drive cancer progression. So far, spatial CNV inference has relied on whole transcriptome-based sequencing technologies. However, advances in image-based spatial transcriptomics (iST) now enable high-plex gene measurement in situ. Here, we introduce an approach that adapts CNV inference to iST data, enabling spatial mapping of malignant clones and the tumor microenvironment, at single-cell resolution. Additionally, we assess how panel size and detection efficiency influence CNV inference.

bioinformatics↗

White matter geometry confounds Diffusion Tensor Imaging Along Perivascular Space (DTI-ALPS) measures

IntroductionThe perivascular space (PVS) is integral to glymphatic function, facilitating fluid exchange and waste clearance in the brain. Diffusion Tensor Imaging Along the Perivascular Space (DTI-ALPS) has been proposed as a non-invasive marker of perivascular diffusion, yet its specificity remains unclear. ALPS measures assume that radial asymmetry in white matter diffusivity predominantly reflects PVS contributions. However, anatomical and microstructural confounds may influence these metrics. MethodsWe systematically evaluated potential biases in ALPS-derived measures using high-resolution, multi-shell diffusion MRI from the Human Connectome Project (HCP) and high-field imaging. Specifically, we examined (1) the prevalence of radial asymmetry across white matter, (2) the influence of crossing fibers on ALPS indices, (3) the impact of axonal undulations and dispersion, and (4) the spatial alignment of vasculature with white matter in ALPS-associated regions. ResultsRadial asymmetry is widespread across white matter and persists even at high b-values, suggesting a dominant contribution from axonal geometry rather than faster PVS-specific diffusion. Crossing fibers significantly inflate ALPS indices, with greater radial asymmetry observed in regions with a greater prevalence of crossing fibers. Furthermore, anisotropic axonal dispersion and undulations introduce systematic asymmetry independent of perivascular diffusion. Finally, high-resolution vascular imaging reveals substantial heterogeneity in medullary vein orientation, challenging the assumption that PVS consistently aligns with the left-right axis in ALPS regions. ConclusionALPS indices are significantly influenced by white matter microstructure, including fiber crossings, undulations, and dispersion. These findings suggest that ALPS-derived metrics may not provide a direct measure of glymphatic function but rather reflect underlying axonal geometry. Interpretations of ALPS-derived metrics as biomarkers of glymphatic function must consider these anatomical complexities, and future studies should integrate advanced modeling approaches to disentangle perivascular contributions from white matter structure.

neuroscience↗

Oligodendroglia vulnerability in the human dorsal striatum in Parkinson's disease

Oligodendroglia are the responsible cells for myelination in the central nervous system and their involvement in Parkinsons disease (PD) is poorly understood. We performed snRNA-seq and image-based spatial transcriptomics of human caudate nucleus and putamen (dorsal striatum) from PD and Control brain donors to elucidate the diversity of oligodendroglia and how they are affected by the disease. We have defined fifteen subclasses, from precursor to mature cells, four of which are disease-associated. These PD-specific populations are characterized by the overexpression of heat shock proteins and distinct expression signatures, including immune responses and myelination alterations. We have also identified disruptions in cell communication and oligodendrocyte development, evidenced by changes in neurotransmitter receptors expression and cell adhesion molecules. These transcriptomic changes correlated with impaired myelin integrity and altered oligodendrocyte distribution in the striatum. Thus, we uncover oligodendroglia as a critical cell type in PD and a potential new therapeutic target.

neuroscience↗

2D, or not 2D? Investigating Vertical Signal Integrity of Tissue Slices

Imaging-based spatially resolved transcriptomics can localize transcripts within tissue sections in 3D. Cell segmentation assigns transcripts to cells and precedes annotation of cell function. However, cell segmentation is usually performed in 2D, thus unable to deal with spatial doublets arising from overlapping cells, resulting in segmented cells containing transcripts originating from multiple cell types. Here we present a computational tool called ovrlpy that identifies overlapping cells, tissue folds, and inaccurate cell-segmentation by analyzing transcript localization in 3D.

bioinformatics↗