Biology subjects

Uhlen, M.

Publications and source records attributed to Uhlen, M..

At least 19 recordsLinked to original sources

bioRxiv2026

The genetic architecture of dementia risk: how Alzheimer's disease vulnerability converges on lipid metabolism and immune cell networks

Although genome-wide association studies (GWAS) have identified numerous dementia risk loci, their cell-type and tissue-specific contexts remain largely unresolved. We introduced HPA GeneSet Explorer, a statistical pipeline designed to systemically map Genome wide association study (GWAS) disease risk genes across the Human Protein Atlas (HPA). This approach generates a multiscale transcriptomic map of genetic risk, spanning systemic organs, brain regions, and cell types. Applying this framework to Alzheimers disease (AD), Lewy body dementia (DLB), and Frontotemporal dementia (FTD) revealed convergent neuronal enrichment in all three diseases. In addition, we identified AD-specific enrichment in immune system and liver associated gene modules, along with DLB-specific enrichment in ciliary modules. By mapping these vulnerabilities in non-demented samples, we provide a blueprint of the baseline vulnerability hotspots that can precede clinical neurodegeneration, offering new targets for disease-specific therapies and biomarker discovery.

neuroscience↗

bioRxiv2026

A cell type-resolved proteomic atlas of the human body

Proteins define what cells do, yet their systematic quantification across human cell types has remained out of reach. Using Deep Visual Proteomics on tissue from a healthy female donor, we built an atlas of 27 cell types across 14 tissues, quantifying two-thirds of all human protein-coding genes, with up to 8,500 per population. The proteome partitions bimodally into a universal core and highly specialized programs. Integration into the Human Protein Atlas Single Cell Resource enabled comparison of RNA and protein abundance at cell type resolution, revealing that concordance depends on pathway rather than cellular identity. This resolution uncovered cancer-testis antigens in oocytes invisible to bulk profiling. Our openly accessible resource provides a foundation for cell type-resolved proteomics in health and disease.

biochemistry↗

bioRxiv2026

Systems-level longitudinal immune profiling reveals individualized immunotypes and genetic associations

The human immune system exhibits substantial inter-individual variation, yet how this variability is coordinated across the system and persists over time remains incompletely defined. Here, we integrated whole-genome sequencing with longitudinal multi-omics immune profiling in 101 healthy individuals followed over two years, combining mass cytometry-based immune cell profiling, PBMC transcriptomics, plasma proteomics, and clinical measurements. Coordinated variation across cellular composition, gene expression, and circulating proteins revealed reproducible immune patterns within individuals across visits. Integrative analyses identified three major immunotypes: adaptive lymphoid (CD4 T cell and B cell enriched), myeloid-inflammation (monocyte and dendritic cell-driven), and cytotoxic (CD8 T cell-dominated), each associated with distinct metabolic and inflammatory profiles. Genome-wide association analyses identified cell-type-specific quantitative trait loci primarily affecting memory lymphocytes, and a polygenic score for memory B cells correlated with both cellular abundance and transcriptional activity. Together, these findings provide a systems-level view for understanding baseline immune heterogeneity in human populations. TeaserLongitudinally stable immune individuality reflects coordinated modules shaped by systemic physiology and genetics

systems biology↗

bioRxiv2026

A high-resolution spatial map of cilia-associated proteins in the human fallopian tube

Molecular alterations in the fallopian tubes play a pivotal role in the development of cancer and reproductive disorders, yet their molecular landscape at the protein level remains poorly defined. Here, we map key fallopian tube proteins at single-cell resolution utilizing an integrated transcriptomics and proteomics approach. Based on RNA-seq analysis we identify 310 genes with elevated expression in fallopian tube, the majority of which are associated with motile cilia function. We spatially characterize 133 of the corresponding proteins in fallopian tube and other human tissues with motile cilia to subcellular structures of ciliated cells, validating the findings with single-cell RNA-seq and mass-spectrometry data. Eleven proteins previously only studied on the transcript level without information in cilia databases are further analyzed in a hydrosalpinx patient, showing a thinner epithelium, lower density of FOXJ1 expression, and reduced expression of FHAD1, RIIAD1, and C2orf81. Our high-resolution spatial map aids in dissecting the pathways underlaying infertility and diseases linked to cilia-specific functions.

molecular biology↗

bioRxiv2026

Systematic Analysis of Human Tissue- and Cell-Specific Metabolic Models Identifies High-Sugar, High Fat Diet Induced Liver Dysregulation

Human tissues exhibit specialized metabolic functions that are essential for maintaining whole-body metabolic homeostasis. To systematically characterize organ- and cell-type-specific metabolic heterogeneity, we constructed 32 tissue-specific and 81 cell-type-specific enzyme-constrained genome-scale metabolic models (ecGEMs) by integrating the global human metabolic network with the tissue- and single-cell transcriptomic data from the Human Protein Atlas (HPA). Our analysis revealed pronounced differences in metabolic network architecture and activity across the human body, identifying key cell types that drive tissue metabolic functions. To demonstrate the applicability of these models, we employed the liver-specific ecGEM to investigate the metabolic reprogramming induced by a high-sugar, high-fat (HSHF) diet, a primary driver of metabolic dysfunction-associated fatty liver disease (MAFLD). Flux balance analysis revealed a fundamental transition in hepatic metabolism: from a flexible, multi-functional system toward a constrained, lipid-centric regime. This state is characterized by carbohydrate and lipid overload, mitochondrial respiratory dysfunction, and a compromised capacity for reactive oxygen species (ROS) detoxification. These computational predictions were validated through integrative analysis of transcriptomic data from a human MAFLD cohort and metabolomic profiles from an in vivo HSHF rat model. Together, this work provides a comprehensive atlas of human metabolic models, enabling the systematic investigation of metabolic features across tissues and conditions from a systems-level perspective. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=142 SRC="FIGDIR/small/706568v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@1015c7org.highwire.dtl.DTLVardef@5c200org.highwire.dtl.DTLVardef@3c982org.highwire.dtl.DTLVardef@a2894d_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗

bioRxiv2025

A high-resolution subcellular map of proteins in ciliated cells

Motile cilia are complex structures regulated by thousands of genes, essential for various physiological functions like respiration and reproduction. Their dysfunction can result in severe conditions like primary ciliary dyskinesia (PCD), highlighting the need for a deeper molecular understanding of their specific ciliary compartments. Interestingly, ciliated cells harbor multiple proteins with limited evidence on biological function, as defined by Functional Evidence (FE) scores, a grading system developed by the Human Proteome Project (HPP). Building upon the stringent antibody validation pipeline of the Human Protein Atlas (HPA) project, we developed a high-throughput workflow that combines a novel multiplex immunohistochemistry protocol with image analysis to investigate protein expression and subcellular localization in motile ciliated cells across five human tissues: nasopharynx, bronchus, fallopian tube, endometrium, and cervix. We spatially mapped >180 proteins, out of which 73% have FE scores 2-5, suggesting that further evidence is needed to establish these proteins biological function. Notably, expression patterns varied between tissues, suggesting that motile cilia proteins are not universally expressed across the different epithelia. Our pipeline constitutes a promising resource for comprehensive mapping of the motile cilia proteome, and a first step towards identifying cilia proteins for functional studies to understand the molecular mechanisms underlying ciliopathies.

cell biology↗

bioRxiv2025

Enhanced Computational and Experimental Approaches for Comparative Analysis of the Human Mycobiome

The mycobiome is now recognized as a critical part of the human microbial ecosystem, playing a significant role in both health and disease. Mycobiome research is currently in its infancy and relies on fungal-specific primers with challenges in bioinformatics to accurately determine the structure of the mycobiota community. In addition, the majority of computational and experimental methods currently in use have been optimized for bacteria rather than for fungi. Here, we provide a comparative analysis of extraction methodologies for metagenome sequencing and subsequent bioinformatic analysis that will enhance fungal species identification. We utilized cultured mock fungal communities, including both yeast and mold species, to evaluate the efficiency of extraction protocols. We further enhanced computational analysis of the mycobiome, using mock and human metagenome data together with our curated catalogue consisting of 984 fungal genomes specific to the human mycobiome. Application of this optimised workflow for oral and gut samples of healthy individuals detected of novel species from Puccinia and Lentinus genera among the established presence of Malassezia, Rhizophagus, Candida and Saccharomyces genera. In addition, Enterocytozoon, was identified specifically in the gut mycobiome. Our pipeline enabled the comprehensive sampling of all fungal genes in a microbial community within a human sample minimising bias, reducing errors and artefacts from amplification, and providing accurate diversity and abundance data for the human mycobiome.

genomics↗

bioRxiv2024

Microglial states determine lesion dynamics in multiple sclerosis

Summary paragraphMultiple sclerosis (MS) is a neuroinflammatory disease of the central nervous system, characterized by demyelinating lesions 1. Lesion expansion contributes to progression and increased disability, while remyelination can recover neurological deficits. However, mechanisms driving lesion dynamics are largely unclear, hindering the development of effective therapeutics. We propose that distinct states of microglia are involved in lesion expansion and remyelination 2,3. Using Stereo-seq, an RNA capture based high-resolution spatial transcriptomics technology with single-cell resolution, on post-mortem human brain tissue, we compared mixed active/inactive lesions with lipid-laden foamy microglia with lesions containing ramified microglia. We identified distinct cellular and molecular mechanisms underlying lesion activity and remyelination, linked to microglia phenotypes and states. Lesions with foamy microglia were characterized by elevated immune activation, increased lymphocyte densities, upregulated immunoglobulin production (IGHG1, IGHG3), increased complement system activity, indication of iron dysregulation (FTL, FTH1), and increased demyelination. In contrast, lesions with ramified microglia exhibited gene expression profiles indicative of myelin stability (ABCA2, QKI) and neuro-axonal protection, fostering an environment conducive to repair and remyelination. Our findings highlight the role of microglial states in lesion expansion and repair in MS and offer promising avenues for the development of therapeutic approaches aimed at preventing MS disability progression.

neuroscience↗

bioRxiv2024

Intrinsic Diversity in Primary Cilia Revealed Through Spatial Proteomics

Primary cilia are a critical organelle found on most human cells, and their dysfunction is linked to hereditary ciliopathies with a wide phenotypic spectrum. Despite their significance, the specific roles of cilia in different cell types remain poorly understood due to limitations in analyzing ciliary protein composition. We employed antibody-based spatial proteomics to expand the Human Protein Atlas to primary cilia. Our analysis identified the subciliary locations of 715 proteins across three cell lines, examining 128,156 individual cilia. We found that 69% of the ciliary proteome is cell-type specific, and 78% exhibited single-cilia heterogeneity. Our findings portray cilia as sensors tuning their proteome to effectively sense the environment and compute cellular responses. We identified 91 novel cilia proteins and found a genetic candidate variant in CREB3 in one clinical case with features overlapping ciliopathy phenotypes. This open, spatial cilia atlas advances research on cilia and ciliopathies.

cell biology↗

bioRxiv2024

A spatiotemporal atlas of human spermatogenesis based on single-cell transcriptomics and multiplex antibody imaging

Understanding how transcriptional programs are translated into functional protein landscapes within intact tissues remains a major challenge in biology. While single-cell RNA sequencing (scRNA-seq) has enabled high-resolution identification of cellular states, corresponding protein expression within its spatial context remains poorly resolved at scale. Here, we present a scalable framework that integrates scRNA-seq with multiplex immunohistochemistry (mIHC) and automated image analysis to achieve spatially resolved, single-cell-level protein mapping across human spermatogenesis. Using this approach, we quantified the expression of nearly 500 proteins across 12 discrete germ cell states within intact human testis tissue, generating a high-resolution spatiotemporal proteomic atlas. Comparative analysis of mRNA and protein expression revealed widespread temporal discordance, indicating that transcriptional state is not always predictive of protein abundance. Notably, we identify delayed translation of key regulators such as PIWIL4, whose protein expression peaks at later differentiation stages than its mRNA expression. Together, our results establish a generalizable strategy for proteome-wide spatial mapping of single-cell states and demonstrate the importance of integrating protein-level measurements to resolve cellular identity and function in complex tissues.

molecular biology↗

bioRxiv2024

Microbiome signatures of virulence in the oral-gut-brain axis influence Parkinson's disease and cognitive decline pathophysiology

The human microbiome is increasingly recognized for its crucial role in the development and progression of neurodegenerative diseases. While the gut-brain axis has been extensively studied, the contribution of the oral microbiome and gut-oral tropism in neurodegeneration has been largely overlooked. Cognitive impairment (CI) is common in neurodegenerative diseases and develops on a spectrum. In Parkinsons Disease (PD) patients, CI is one of the most common non-motor symptoms but its mechanistic development across the spectrum remains unclear, complicating early diagnosis of at-risk individuals. Here, we generated 228 shotgun metagenomics samples of the gut and oral microbiomes across PD patients with either mild cognitive impairment (PD-MCI) or dementia (PDD), and a healthy cohort, to study the role of the gut and oral microbiomes on CI in PD. In addition to revealing compositional and functional signatures, the role of pathobionts, and dysregulated metabolic pathways of the oral and gut microbiome in PD-MCI and PDD, we also revealed the importance of oral-gut translocation in increasing abundance of virulence factors in PD and CI. The oral-gut virulence was further integrated with saliva metaproteomics and demonstrated their potential role in dysfunction of host immunity and brain endothelial cells. Our findings highlight the significance of the oral-gut-brain axis and underscore its potential for discovering novel biomarkers for PD and CI.

microbiology↗

bioRxiv2024

Dissecting autonomous enzyme variability in single cells

Metabolic enzymes perform life-sustaining functions in various cellular compartments. Anecdotally, metabolic activity is observed to vary between genetically identical cells, which impacts drug resistance, differentiation, and immune cell activation. However, no large-scale resource systematically reporting metabolic cellular heterogeneity exists. Here, we leverage imaging-based single-cell spatial proteomics to reveal the extent of non-genetic variability of the human enzymatic proteome, as a proxy for metabolic states. Nearly two fifths of enzymes exhibit cell-to-cell variable expression, and half localize to multiple cellular compartments. Metabolic heterogeneity arises largely autonomously of cell cycling, and individual cells reestablish these myriad metabolic phenotypes over several cell divisions. Multiplexed imaging revealed that metabolic states are continuous and that the correlation between metabolic pathways is metabolic state dependent. These results establish cell-to-cell enzymatic heterogeneity as an organizing principle of cell biology that may rewire our understanding of drug resistance, treatment design, and other aspects of medicine.

cell biology↗

bioRxiv2024

Multi-tissue network analysis reveals the effect of JNK inhibition on dietary sucrose-induced metabolic dysfunction in rats

Excessive consumption of sucrose, in the form of sugar-sweetened beverages, has been implicated in the pathogenesis of metabolic dysfunctionLassociated fatty liver disease (MAFLD) and other related metabolic syndromes. The c-Jun N-terminal kinase (JNK) pathway plays a crucial role in response to dietary stressors, and it was demonstrated that the inhibition of the JNK pathway could potentially be used in the treatment of MAFLD. However, the intricate mechanisms underlying these interventions remain incompletely understood given their multifaceted effects across multiple tissues. In this study, we challenged rats with sucrose-sweetened water and investigated the potential effects of JNK inhibition by employing network analysis based on the transcriptome profiling obtained from hepatic and extrahepatic tissues, including visceral white adipose tissue, skeletal muscle, and brain. Our data demonstrate that JNK inhibition by JNK-IN-5A effectively reduces the circulating triglyceride accumulation and inflammation in rats subjected to sucrose consumption. Coexpression analysis and genome-scale metabolic modelling reveal that sucrose overconsumption primarily induces transcriptional dysfunction related to fatty acid and oxidative metabolism in the liver and adipose tissues, which are largely rectified after JNK inhibition at a clinically relevant dose. Skeletal muscle exhibited minimal transcriptional changes to sucrose overconsumption but underwent substantial metabolic adaptation following the JNK inhibition. Overall, our data provides novel insights into the molecular basis by which JNK inhibition exerts its metabolic effect in the metabolically active tissues. Furthermore, our findings underpin the critical role of extrahepatic metabolism in the development of diet-induced steatosis, offering valuable guidance for future studies focused on JNK-targeting for effective treatment of MAFLD.

bioinformatics↗

bioRxiv2024

Longitudinal Exposomics in a Multiomic Wellness Cohort Reveals Distinctive and Dynamic Environmental Chemical Mixtures in Blood

Chemical exposomes can now be comprehensively measured in human blood, but robust application of chemical exposomics in cohort studies requires knowledge of the longitudinal stability and interindividual variability of exogenous molecular profiles. Here we applied chemical exposomics to plasma of 46 adults, each sampled six times over two years in a multiomic wellness cohort. New chemicals were discovered, distinctive co-exposure patterns were observed, and intra-class correlation coefficients (ICC) for 519 confidently annotated substances are reported to support study design. Longitudinal stability of the chemical exposome (mean ICC 0.30) was significantly lower than the proteome, metabolome, lipidome or microbiome, and must be measured more frequently than other molecular profiles in health studies. Mixed-effects models nevertheless revealed significant associations between testosterone and perfluoroalkyl substances, and significant time-trends for low and high stability exposures alike. Complex exposome data structures were visualized and explored, demonstrating great potential for longitudinal exposomics in precision health research. TeaserThe first cohort-level application of longitudinal exposomics revealed novel and dynamic co-exposures in blood of relevance to precision health.

pharmacology and toxicology↗

bioRxiv2023

Expanding the GPCR-RAMP interactome

Receptor activity-modifying proteins (RAMPs) can form complexes with G protein-coupled receptors (GPCRs) and regulate their cellular trafficking and pharmacology. RAMP interactions have been identified for about 50 GPCRs, but only a few GPCR-RAMP complexes have been studied in detail. To elucidate a complete interactome between GPCRs and the three RAMPs, we developed a customized library of 215 Dual Epitope-Tagged (DuET) GPCRs representing all GPCR subfamilies. Using a multiplexed suspension bead array (SBA) assay, we identified 122 GPCRs that showed strong evidence for interaction with at least one RAMP. We screened for native interactions in three cell lines and found 23 GPCRs that formed complexes with RAMPs. Mapping the GPCR-RAMP interactome expands the current system-wide functional characterization of RAMP-interacting GPCRs to inform the design of selective GPCR-targeted therapeutics. One-Sentence SummaryNovel complexes between G protein-coupled receptors and interacting proteins point to a system-wide regulation of GPCR function.

biochemistry↗

bioRxiv2023

Pathogenic entero- and salivatypes harbour changes in microbiome virulence and antimicrobial resistance genes with increasing chronic liver disease severity

Background & AimsLife-threatening complications of cirrhosis are triggered by bacterial infections, with the ever-increasing threat of antimicrobial resistance (AMR). Alterations in the gut microbiome in decompensated cirrhosis (DC) and acute-on-chronic liver failure (ACLF) are recognised to influence clinical outcomes, whilst the role of the oral microbiome is still being explored. Our aims were to simultaneously interrogate the gut and oral micro- and mycobiome in cirrhotic patients, and assess microbial community structure overlap in relation to clinical outcomes, as well as alterations in virulence factors and AMR genes. Methods18 healthy controls (HC), 20 stable cirrhotics (SC), 50 DC, 18 ACLF and 15 with non-liver sepsis (NLS) i.e. severe infection but without cirrhosis, were recruited at a tertiary liver centre. Shotgun metagenomic sequencing was undertaken from saliva (S) and faecal (F) samples (paired where possible). Salivatypes and enterotypes based on clustering of genera were calculated and compared in relation to cirrhosis severity and in relation to specific clinical parameters. Virulence and antimicrobial resistance genes (ARGs) were evaluated in both oral and gut niches, and distinct resistotypes identified. ResultsSpecific saliva- and enterotypes revealed a greater proportion of pathobionts with concomitant reduction in autochthonous genera with increasing cirrhosis severity, and in those with hyperammonemia. Overlap between oral and gut microbiome communities was observed and was significantly higher in DC and ACLF vs SC and HCs, independent of antimicrobial, beta-blocker and acid suppressant use. Two distinct gut microbiome clusters [ENT2/ENT3] harboured genes encoding for the phosphoenolpyruvate:sugar phosphotransferase system (PTS) system and other virulence factors in patients with DC and ACLF. Substantial numbers of ARGs (oral: 1,218 and gut: 672) were detected with 575 ARGs common to both sites. The cirrhosis resistome was significantly different to HCs, with three and four resistotypes identified for the oral and gut microbiome, respectively. DiscussionOral and gut microbiome profiles differ significantly with increasing severity of cirrhosis, with progressive dominance of pathobionts and loss of commensals. DC and ACLF have significantly worse microbial diversity than NLS, despite similar antimicrobial exposure, supporting the additive patho-biological effect of cirrhosis. The degree of microbial community overlap between sites, frequency of virulence factors and presence of ARGs, all increment significantly with hepatic decompensation. These alterations may predispose to higher infection risk, poorer response to antimicrobial therapy and worsening outcomes, and provide the rationale for developing non-antibiotic-dependent microbiome-modulating therapies.

physiology↗

bioRxiv2023

A human stomach cell type transcriptome atlas

The identification of cell type-specific genes and their modification under different conditions is central to our understanding of human health and disease. The stomach, a hollow organ in the upper gastrointestinal tract, provides an acidic environment that contributes to microbial defence and facilitates the activity of secreted digestive enzymes to process food and nutrients into chyme. In contrast to other sections of the gastrointestinal tract, detailed descriptions of cell type gene enrichment profiles in the stomach are absent from the major single cell sequencing-based atlases. Here, we use an integrative correlation analysis method to predict human stomach cell type transcriptome signatures using unfractionated stomach RNAseq data from 359 individuals. We profile parietal, chief, gastric mucous, gastric enteroendocrine, mitotic, endothelial, fibroblast, macrophage, neutrophil, T-cell and plasma cells, identifying over 1600 cell type-enriched genes. We uncover the cell type expression profile of several non-coding genes strongly associated with the progression of gastric cancer and, using a sex-based subset analysis, uncover a panel of male-only chief cell-enriched genes. This study provides a roadmap to further understand human stomach biology.

systems biology↗

bioRxiv2023

A tissue centric atlas of cell type transcriptome enrichment signatures

Genes with cell type specific expression typically encode for proteins that have cell type specific functions. Single cell RNAseq (scRNAseq) has facilitated the identification of such genes, but various challenges limit the analysis of certain cell types and lowly expressed genes. Here, we performed an integrative network analysis of over 6000 bulk RNAseq datasets from 15 human organs, to generate a tissue-by-tissue cell type enrichment prediction atlas for all protein coding genes. We profile all the major constituent cell types, including several that are fragile or difficult to process and thus absent from existing scRNAseq-based atlases. The stability and read depth of bulk RNAseq data, and the high number of biological replicates analysed, allowed us to identify lowly expressed cell type enriched genes that are difficult to classify using existing methods. We identify co-enriched gene panels shared by pancreatic alpha and beta cells, chart temporal changes in cell enrichment signatures during spermatogenesis, and reveal that cells in the hair root are a major source of skin enriched genes. In a cross-tissue analysis, we identify shared gene enrichment signatures between highly metabolic and motile cell types, and core identity profiles of cell types found in across tissue types. Our study provides the only cell type gene enrichment atlas generated independently of scRNAseq, representing a new addition to our existing toolbox of resources for the understanding of gene expression across human tissues.

systems biology↗