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Gupta, P.

Publications and source records attributed to Gupta, P..

At least 37 records · Page 2Linked to original sources

A scalable approach to resolving variants of uncertain significance

Over 90% of missense variants across [~]4,000 disease-associated genes are variants of uncertain significance (VUS). Experimental variant effect measurements provide critical evidence about pathogenicity and inform disease biology, but most variants lack data and clinical translation has been limited. The Impact of Genomic Variation on Function Consortium generated experimental data for 62,215 variants across ten genes using multiplexed assays and 1,407 variants across 163 genes using arrayed assays, curated 193,139 additional community-generated variant effect measurements across 30 additional genes, and developed automated calibration methods for translating experimental data and variant effect predictions into clinical evidence. To reduce current VUS, we developed a scalable workflow using only experimental and predictive evidence, enabling reclassification of 75% of the 16,115 VUS in these genes as pathogenic or benign with <1% error. To minimize future VUS, we analyzed >90,000 unobserved variants; 62% had enough evidence to be "preclassified" as pathogenic or benign. We validated our data, evidence and classifications using All of Us and created interactive resources to enable clinical use of the calibrated data. Thus, for 40 genes, representing 1% of the clinical genome, we resolve most existing VUS and future variants, illustrating how systematic use of scalable evidence can empower genomic medicine.

genomics↗

European taurine vs Indian indicine cattle: a comparative genomic study reveals regions of differentiation during evolution and selection during crossbreeding

Cattle are broadly classified into two subspecies: Bos taurus, adapted to temperate climates, and Bos indicus, adapted to tropical environments. Indicine cattle show better heat tolerance and stronger disease resistance, whereas taurine cattle are known for higher milk yield and better meat quality. Improving milk yield while maintaining resistance to heat stress and infectious diseases is an important objective in cattle breeding. Marker-assisted selection is an effective approach for improving economically important traits, highlighting the need to understand genetic differences between taurine and indicine cattle. However, genome-wide comparisons between European taurine and Indian indicine cattle remain limited. To address this gap, whole-genome sequencing of 48 Indian indicine cattle was performed and combined with publicly available data. This enabled a comparative genomic analysis of 74 Indian indicine and 83 European taurine individuals to identify genes involved in heat tolerance and immune response. Genome-wide analyses using Fst and XP-CLR identified 4,343 and 1,457 differentiated genes, respectively, with 826 genes common to both methods. These genes were mainly associated with immune response, protein stability, and cytoskeletal structure. Strong selection signals were observed in three heat shock protein genes (DNAJC11, DNAJC5, and DNAJB11) and 229 immune-related genes. To examine the inheritance of these genes through crossbreeding, a haplotype-resolved genome assembly was generated for the Indian crossbreed Sunandini, which showed predominantly taurine ancestry (69.13-96.04%), with a smaller indicine contribution (1.22-1.87%). Several genes related to heat tolerance and immune response were inherited exclusively from indicine cattle, highlighting their importance for environmental adaptation and future breeding programs.

genomics↗

An integrated, scaled approach to resolve TSC2 variants of uncertain significance

Obtaining a precise genetic tuberous sclerosis diagnosis is a challenge as many missense TSC2 variants are variants of uncertain significance (VUS). VUS in TSC2 have been resolved by one-at-a-time functional assays, but these assays cannot scale to the 3,634 TSC2 missense VUS observed so far. To address this challenge, we used massively parallel sequencing to measure the steady-state abundance of almost 9,000 TSC2 missense variants and developed an mTOR pathway activity assay using genome editing and cell sorting to generate activity scores for 391 missense variants. 1,288 of 8,891 (14.49%) missense variants assayed had altered TSC2 abundance, and 69 of 391 (17.65%) missense variants assayed had altered mTOR pathway activity. Calibration and integration of these data into classification of variants identified in a clinical cohort putatively reclassified 212 of 276 (76.8%) TSC2 missense VUS. These datasets will lead to improved genetic diagnosis of tuberous sclerosis with potential positive impacts on the clinical management of patients and their families.

genetics↗

Designing AI-programmable therapeutics with the EDEN family of foundation models

The ability to interpret, modify, and design DNA has driven many of the most significant advances in modern medicine, from diagnostics, biologics, and vaccines to cell and gene therapies. However, the inherent complexity of biological systems means that most modern medicines are still engineered using bespoke, labor-intensive processes. To address the need for a generalisable and programmable approach to therapeutic design, we introduce the EDEN (environmentally-derived evolutionary network) family of metagenomic foundation models, including a 28 billion parameter model trained on 9.7 trillion nucleotide tokens from BaseData1. This dataset, at the time of training, contained more than 10 billion novel genes from over 1 million new species, and is intentionally enriched for environmental and host-associated metagenomes, phage sequences, and mobile genetic elements, enabling the model to learn from diverse and novel cross-species evolutionary mechanisms and apply them to key challenges in human health. EDEN achieves state-of-the-art performance across a series of predictive and generative genomic and protein benchmarks. To demonstrate the models broad applicability across biology, we evaluate EDENs capacity for programmable therapeutic design by challenging a single architecture to design biological novelty across three distinct therapeutic modalities, disease areas and biological scales: (i) large gene insertion, (ii) antibiotic peptide design, and (iii) microbiome design. First, we demonstrate AI-programmable Gene Insertion (aiPGI), in which EDEN designs de novo large serine recombinases (LSRs) capable of inserting large pieces of DNA at desired target sites in the human genome when prompted only on 30 nucleotides of DNA sequence from the desired target site. In low-N experimental validation, EDEN generated multiple active recombinases for all tested disease-associated genomic loci (ATM, DMD, F9, FANCC, GALC, IDS, P4HA1, PHEX, RYR2, USH2A) and 4 potential safe harbor sites in the human genome. EDEN achieves an overall functional hit rate of 63.2% across diverse DNA prompts when prompted on only 30bp of DNA from outside the training data. 50% of EDEN-generated LSRs were active in human cells, achieving therapeutically relevant levels of CAR insertion in primary human T cells. We also show that EDEN can generate active bridge recombinases when prompted on the associated guide RNA alone, with sequence identities to training and public data as low as 65%. These results pave the way for a new generation of cell and gene therapies by opening the door to rapid, programmable and site-specific integration of large genetic payloads without double-strand breaks. This offers an alternative to the safety, efficiency and payload limitations inherent in viral or nuclease-based editing at thousands of currently intractable human therapeutic targets. Second, we use the same model to generate a focused low-N library of novel antimicrobial peptides where 97% showed activity, with top candidates achieving single-digit micromolar potency against critical-priority multidrug-resistant pathogens. Third, to demonstrate that EDEN captures inter-genomic features, we design a gigabase-scale microbiome with over 94,000 synthetic metagenomic assemblies, including prophage genomes and correct cross-species metabolic pathway completions. The EDEN-generated synthetic microbiome covers 9,067 species with a biome-specific taxonomic accuracy of 99%. Over 1,500 of the generated species were outside the fine-tuning dataset while retaining the correct microecological properties and biome association, thus significantly expanding genetic and taxonomic diversity. Together, these results establish a new strategic direction for AI-programmable therapeutics, in which a single foundation model architecture designs candidate therapeutics across diverse modalities and disease areas. This suggests that the combination of billions of years of evolutionary data with specific therapeutic records offers a clear, scaling-driven path to making therapeutic design a predictable engineering discipline. O_FIG O_LINKSMALLFIG WIDTH=141 HEIGHT=200 SRC="FIGDIR/small/699009v2_ufig1.gif" ALT="Figure 1"> View larger version (59K): org.highwire.dtl.DTLVardef@68c20borg.highwire.dtl.DTLVardef@19b8e8corg.highwire.dtl.DTLVardef@1ab9362org.highwire.dtl.DTLVardef@1592cb7_HPS_FORMAT_FIGEXP M_FIG C_FIG

genomics↗

Root-Suppressed Phenotype of Tomato Rs Mutant is Seemingly Related to Expression of Root-Meristem-Specific Sulfotransferases

Root development and growth are governed by a complex interplay between a plants genetic makeup and its ambient environment. In this study, we characterised a radiation-induced root-suppressed (Rs) mutant mapped on chromosome 4 of tomato. Rs seedlings had a dwarf stature, and mature plants exhibited abnormalities in their leaves, flowers, and fruits. The partial rescue of seedling root elongation by H2S donors or L-cysteine hinted that its root defect is linked to disrupted sulfur metabolism. The metabolite profiling of organs revealed altered homeostasis, analogous to that of plants subjected to sulfur starvation. Whole-genome sequencing revealed that the Rs genome was replete with genetic lesions; therefore, the pleiotropic morphological abnormalities likely emanate from mutations in other genes, including those related to sulfur. Among S-related genes, two sulfotransferase genes (Solyc04g028380 and Solyc04g028390) localised on chromosome 4 showed mutations in their promoters. The expression of both sulfotransferases is confined to the root meristem, suggesting a causal connection with the suppression of root growth. These sulfotransferases belonged to a distinct subset exclusively expressed in the root meristem and were distinct from the tyrosine sulfotransferase involved in the sulfation of root growth factors. Our study suggests that a sulfur-related module, other than tyrosine sulfotransferase, may regulate root development.

plant biology↗

Slow Spindle Trains During Daytime Naps are Associated with Improved Declarative Memory Consolidation

Memory consolidation refers to the process by which newly encoded memories are strengthened and retained over time, and ample evidence indicates that sleep supports this process for both procedural and declarative memories. Although sleep spindles during non-rapid eye movement (NREM) sleep have been associated to consolidation, it remains unclear whether all spindle types contribute equally. Spindles vary in frequency and topography-slow spindles ([&le;]12.5Hz) predominating over frontal regions, whereas fast spindles (>12.5Hz) peak parietally - and recent work suggests that procedural memory consolidation during overnight sleep is related to the temporal organization of spindles in trains (i.e., events occurring <6s apart). Here we investigated whether a similar mechanism operates for declarative memory during daytime naps. Participants were assigned to a Nap (N=23) or No-Nap (N=15) group, and completed an object-spatial location task involving 36 item-location associations. Memory was assessed immediately after learning and again following a 90-minute nap or an equivalent wake period. Results showed that the Nap group exhibited significantly better delayed memory, as measured by combined recall-recognition score, and a greater proportion of participants maintained or improved their performance. In the Nap group, memory performance correlated with local spindle density at frontal and parietal sites, and, critically, with the proportion of slow spindles clustered in trains during NREM2. These findings suggest the temporal organization of slow spindles into clusters support declarative memory consolidation, pointing to a shared spindle-based mechanism across domains.

neuroscience↗

WRN helicase regulates mitophagy by resolving an intricate nexus of G-quadruplexes-R loops-ATG7 pre-mRNA maturation in cancer

Werner syndrome (WS) is a progeroid and cancer-predisposition disorder caused by loss of the Werner RECQ helicase-exonuclease (WRN), a key genome-maintenance enzyme essential for replication-stress signalling and DNA-repair. WS patients also develop metabolic abnormalities, including fatty-liver and diabetes, suggesting a link between WRN-deficiency and mitochondrial-dysfunction. WRN is frequently epigenetically silenced in cancers, yet its precise role in mitochondrial homeostasis in cancer remains unclear. Here, we define a role for WRN in regulating mitophagy and autophagy in cancer. WRN-deficient cells show defective mitochondrial respiration, morphology, and mitophagosome/autophagosome-maturation under basal and cisplatin-induced stress. Mechanistically, WRN-loss causes strong reduction of ATG7-protein, compromising autophagosome-biogenesis. Chromatin immunoprecipitation reveals accumulation of unresolved G-quadruplex structures (G4-DNA) across the ATG7-locus in WRN-deficient cells. Paradoxically, ATG7-mRNA expression is elevated despite reduced ATG7-protein in WRN-deficient cells, indicating a post-transcriptional defect. Further, we show that WRN resolves G4-DNA which prevent R-loops formation and interacts with the mRNA-processing factor U2AF35, independent of its helicase-exonuclease functions, to promote maturation of nascent ATG7 transcripts. In cancer patients, WRN level also inversely correlated with post-transcriptional defects in ATG7 mRNA. Collectively, our findings suggest pivotal association of WRN-loss in autophagy fidelity, which may further contribute to oncogenic transformation in WRN-deficient tissues and exacerbate cancer susceptibility in WS-patients. Significance statementWRN is well recognized for its roles in DNA repair and genome maintenance, which are essential for suppressing tumorigenesis and Werner syndrome (WS)-associated premature-aging. However, its functions in mitochondrial regulation remain underexplored, despite WS patients exhibiting severe metabolic defects and increased cancer risk. Here, we uncover a mechanistic link between WRN and autophagy/mitophagy, showing that WRN resolves G-quadruplexes and R-loops to enable proper post-transcriptional processing and translation of ATG7, a key autophagy enzyme. WRN-loss associated defective-autophagy may heighten the initiation of tumorigenesis in both WS-patients and in normal individual with mutated WRN in different tissue-types. As WRN is actively pursued as a synthetic-lethal target and multiple WRN inhibitors progress through clinical-development, our findings highlight mitochondrial quality-control defects as an additional determinant of WRN-targeted therapeutic-response.

cell biology↗

Compaction and swelling of single stretched DNAs driven by molecular crowding

We present a theory for the effects of osmotic pressure exerted by macromolecular crowders, on a double-stranded DNA or other semiflexible polymer extended by tension. Our results predict the force and crowder-density dependence of the polymer extension. The lowest-order effect is a crowder-dependent compression that counteracts the external stretching force, which can collapse the polymer at a crowder-dependent critical force f*. This compression is dependent on crowder radius r and density{phi} , with higher densities and smaller radii having greater effects on the force-extension curve. At first order in perturbation theory we also find a fluctuation-dependent correction to the depletion volume, which can overwhelm the simple compression effect for large crowders, leading to expansion of the polymer.

biophysics↗

A clinical-stage oncology compound selectively targets drug-resistant cancers

Re-evaluating existing clinical compounds can uncover previously unrecognized mechanisms that reshape a drugs therapeutic potential. The small molecule Procaspase-Activating Compound 1 (PAC-1) entered oncology testing as a proposed activator of caspase-driven apoptosis. Here, we show that PAC-1-driven cytotoxicity occurs in the absence of executioner caspase expression, demonstrating that its anti-cancer activity occurs via an alternative mechanism. We provide genetic, biochemical, and biophysical evidence demonstrating that PAC-1 functions as a highly selective iron chelator that eliminates cancer cells by disrupting iron homeostasis. Unexpectedly, we discovered that expression of the key chemotherapy-resistance pump MDR1 confers marked hypersensitivity to PAC-1 treatment. While PAC-1 is only weakly effluxed by MDR1 under basal conditions, this process is potentiated when PAC-1 is bound to iron. Consequently, PAC-1 induces progressive iron depletion and selective cytotoxicity in otherwise drug-resistant MDR1-expressing cancer cells. Together, these findings redefine PAC-1s mechanism-of-action and establish a framework for exploiting multidrug resistance as a therapeutic vulnerability through targeted iron starvation.

cancer biology↗

Aridity drives global convergence of desert microbiomes and biogeochemical activities

Deserts cover a third of the worlds surface, supporting unique biomes and ecosystem services. Yet, we lack a comprehensive assessment of what defines and drives the microbial communities that dominate life in these regions. Here, we conducted a standardized field survey in contrasting cold, hot, and polar deserts across the seven continents, and observed geographically distant deserts share similar structure, function, and activities. Desert communities are dominated by genomically streamlined Actinobacteriota and Chloroflexota, and compared with non-desert soils, are significantly enriched with stress tolerance genes, mobile genetic elements, and antiviral strategies, revealing previously unknown ecological and evolutionary dynamics. Metabolically, these communities exhibit reduced capacity for carbohydrate and protein degradation, and instead are enriched for chemosynthetic carbon fixation, continuous energy harvesting using atmospheric trace gases and sunlight, and energy reserve biosynthesis. All sampled soils mediated respiration, trace gas oxidation, and carbon fixation, with detectable activity even in hyper-arid Atacama and Antarctic soils at the margins of life. Driver analyses identified aridity as the primary overriding driver of the microbial communities and biogeochemical activities. Collectively, these findings suggest that aridity selects for metabolically self-sufficient taxa capable of continuously meeting energy and carbon needs independently of vegetation-derived inputs, while enduring physicochemical stressors and potentially elevated viral pressure. These new insights are integral to forecast the future of soils amid increasing desertification. Significance statementDesert soils occupy a vast and expanding portion of Earth, yet what defines and governs their dominant microbial life remains incompletely defined. By assessing the composition, capabilities, and activities of microbial communities across deserts on all seven continents, we identify unifying signatures of life under extreme water limitation. We show microbial communities are highly self-sufficient, capable of acquiring energy and carbon even where plant inputs are minimal. This planetary-scale understanding of the desert microbiome has important ramifications for forecasting potential shifts of microbial communities and the services they provide as desertification intensifies.

microbiology↗

Interaction and co-phase separation of SARS-CoV-2 nucleocapsid protein and human hnRNPA1 and its implications for viral life cycle

The nucleocapsid (N) protein of SARS-CoV-2 is central to viral assembly and replication. It binds viral RNA to form a helical nucleocapsid and enabling genome packaging and release into host cells. Human hnRNPA1, one of the most abundant RNA-binding proteins in eukaryotes, regulates key aspects of RNA metabolism, including splicing, transcription, localisation, and transport. Here, we report a direct physical interaction between SARS-CoV-2 N protein and hnRNPA1 with moderate affinity (Kd [~] 0.18 M), primarily mediated through their intrinsically disordered regions. Furthermore, we found that these proteins co-phase separate in vitro and co-localise within stress granules in cells. In vivo studies reveal that hnRNPA1 suppresses viral replication, suggesting that the N protein - hnRNPA1 interaction plays an important role in modulating the viral life cycle.

biochemistry↗

The EXS domain of rice PHOSPHATE1 (OsPHO1;2) reduces the low phosphate response through jasmonate signaling

Phosphorus (P) is a vital macronutrient essential for plant growth, and its deficiency significantly hampers agricultural productivity. The PHOSPHATE 1 (PHO1) protein family, characterized by an N-terminal SPX domain, four transmembrane (4TM) domains, and a C-terminal EXS domain, is pivotal in transporting phosphate (Pi) from roots to shoots. Rice, PHO1;2 plays a crucial role in the Pi export process, and defects in this gene cause severe growth retardation and Pi deficiency symptoms even when external Pi levels are adequate. This study examined the roles of the EXS domain and the combined 4TM+EXS domains of OsPHO1;2 in supporting plant growth responses, independent of Pi transport activity, as well as their influence on hormone signaling and gene regulation. Using CRISPR/Cas9, rice lines expressing specific OsPHO1;2 domains (EXS or 4TM+EXS) were created by targeted deletion of particular domain-coding regions. Phenotypic analysis under Pi-sufficient and deficient conditions, as well as phosphate profiling, revealed that EXS lines exhibited notably better growth than loss-of-function mutants, ospho1;2, during early development, despite having similar shoot Pi levels to the null mutant. These lines showed lower levels of defense hormones (jasmonic acid) than ospho1;2 but were comparable to those in the wild type. Conversely, 4TM+EXS lines exhibited growth patterns similar to ospho1;2 mutants. RNA sequencing indicated that the phosphate starvation response (PSR) and defense pathways were less pronounced in the EXS lines compared to ospho1;2 mutants. However, both EXS and 4TM+EXS lines showed seed development defects and reduced total phosphorus content in seeds, mirroring the ospho1;2 phenotype. Heterozygous plants carrying one functional OsPHO1;2 allele displayed normal growth and seed development, indicating the mutations recessive nature. The findings suggest that the EXS domain of OsPHO1;2 can promote plant growth independently of Pi transport by decreasing PSR and modulating defense hormone pathways. This further suggests a signaling role for PHO1 domains beyond direct Pi translocation. Overall, these results enhance our understanding of Pi homeostasis and may help form strategies for breeding P-efficient crops.

molecular biology↗

A Reference Atlas of the Human Dorsal Root Ganglion

Somatosensory perception largely emerges from diverse peripheral sensory neurons whose cell bodies reside in dorsal root ganglia (DRG). Damage or dysfunction of DRG neurons is a major cause of chronic pain and sensory loss. In mice, deep single-cell transcriptomic profiling and genetically defined models have offered important clues into DRG function, but in humans, the cellular and molecular landscape of DRG neurons remains less understood. Here, we constructed a reference cell atlas of the human DRG by profiling transcriptomes of cells and nuclei from 126 donors sampled across cervical, thoracic, and lumbar DRGs. This atlas resolves 22 neuronal subtypes, including known and previously unrecognized subtypes linked to nociception, mechanosensation, thermosensation, and proprioception, as well as 10 types of non-neuronal cells. Cross-species integration, spatial transcriptomics, and microneurography enabled cell-type-specific comparisons of soma size and conduction velocity between species. Human DRG somata are larger across all cell types than their mouse counterparts, and the conduction velocities of human hair follicle innervating A-fibers are faster than in mice, suggesting a functional shift in rapid mechanical detection in humans. This integrated human DRG reference cell atlas provides a resource for exploring new molecular and physiological features of human DRG, which could help identify new strategies for treating chronic pain and other diseases of the peripheral nervous system.

neuroscience↗

Cdh-2 modulates tissue stiffness and cortical F-actin distribution in neuroepithelial cells to drive forebrain roof plate invagination.

The invagination of the dorsal forebrain roof plate is critical for cerebral hemisphere formation. Although mutations in several genes have been linked to holoprosencephaly (HPE), and studies in chick embryos have implicated key signaling pathways, the mechanisms by which mechanical forces drive roof plate invagination remain poorly understood. To address this gap, we employed atomic force microscopy to map the spatiotemporal changes in tissue stiffness across the dorsal forebrain in the chick embryo during roof plate invagination. Our analysis revealed that neuroepithelium that is uniformly stiff initially, undergoes dramatic remodelling during this period of morphological change. Ultimately a pronounced stiffness gradient emerges, with the roof plate midline becoming markedly more compliant than the dorsolateral regions. Through expression screening, we identified Cdh2 as a candidate molecular regulator, whose spatiotemporal expression pattern closely mirrors the observed stiffness gradient. Mechanistically, we found that the interplay between Cdh2 and F-actin modulates tissue stiffness by regulating Cdh2 expression levels, apical adherens junction stability, and cortical F-actin distribution along the apico-basal axis of neuroepithelial cells. These findings underscore the importance of apico-basal localization of Cdh2 and provide critical insights into the mechanical forces and molecular interactions that govern forebrain roof plate morphogenesis, while also shedding light on the pathogenesis of HPE.

developmental biology↗

ZEB1 Is a Key Regulator of Cardiomyocyte Structure and Function

BackgroundOur previous work identified the transcription factor "Zinc Finger E-Box Binding Homeobox 1" (ZEB1) as a downstream effector of Cytoplasmic Polyadenylation Element Binding Protein 4 (CPEB4), an RNA-binding protein responsive to cardiac stress. While ZEB1 is known for its role in cancer metastasis and epithelial-to-mesenchymal transition (EMT), its function in cardiomyocytes is not well understood. Based on previous findings, we hypothesize that ZEB1 is essential for maintaining the structural integrity and mitochondrial function of cardiomyocytes. MethodsAAV9-Zeb1 was used for the overexpression of Zeb1. Using a myosin heavy chain alpha (MHC) Cre system, we created a Zeb1 conditional knockout mouse. To evaluate cardiac function and structure, we used echocardiography, electron microscopy, immunohistochemistry. We identified differentially expressed genes following Zeb1 deletion using RNA-seq and determined direct Zeb1 target genes by integrating this transcriptome data with a Zeb1 ChIP-seq dataset. ResultsZEB1 deletion leads to sarcomere damage, mitochondrial dysfunction, and dedifferentiation, with more pronounced effects in females. Overexpression promotes hypertrophic remodeling. Echocardiographic analysis showed progressive systolic dysfunction, and histology revealed sarcomeric disarray, again especially in females. A tamoxifen-inducible ZEB1 knockout mouse model only confirmed ZEB1s crucial role in fully differentiated cardiomyocytes in female mice. Integrated analysis of RNA-seq and ChIP-seq revealed that Zeb1 directly regulated mitochondrial genes, thereby playing a critical role cardiomyocyte energy supply and having secondary effects on cardiac structure and function. ConclusionsZEB1 is critical for cardiomyocyte homeostasis, and maintaining its function is necessary for normal cardiac performance and structure.

molecular biology↗

Mapping the miRNA Landscape of Gallbladder Cancer: Genome-Wide Insights and Functional Implications

Gallbladder cancer (GBC), the sixth most aggressive gastrointestinal cancer, exhibits complex molecular dysregulation involving microRNAs (miRNAs) that drive tumor progression. This study has comprehensively profiled genome-wide miRNA expression in GBC cells revealing 971 known miRNAs, and 27 novel miRNAs. We identified 388 significantly differentially expressed (DE) miRNAs (191 upregulated and 197 downregulated), providing valuable insights into the various complex molecular mechanisms underlying GBC pathogenesis. These DE miRNAs target key biological processes, including hormone-induced signaling pathways, cell adhesion molecules, apoptosis, ferroptosis, and cancer-associated pathways, particularly those linked to gastric cancer. Profiling of DE miRNAs in GBC cells presents opportunities for novel miRNA-based diagnostic and prognostic approaches in GBC. Notably, among the DE miRNAs, the miR-17[~]92 cluster was significantly suppressed in GBC cells. We also show that miR-17[~]92 cluster targets and destabilizes an S-phase licensing factor Cdt2 (an oncogene) in GBC cells. miR-17[~]92 mediated negative regulation of Cdt2 induces S-phase arrest, leading to checked proliferation and invasive capabilities of GBC cells specifically, without any significant effect on non-cancerous cells, presenting miR-17[~]92 as a potential miRNA therapeutic for GBC. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=191 SRC="FIGDIR/small/679165v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@243970org.highwire.dtl.DTLVardef@36f578org.highwire.dtl.DTLVardef@2dba37org.highwire.dtl.DTLVardef@1c9e1b_HPS_FORMAT_FIGEXP M_FIG Graphical Abstract C_FIG

cancer biology↗

Single nucleus multiomic atlas of human dorsal root ganglia reveals the contribution of non-neuronal cell types to pain

Sensory neurons residing in dorsal root ganglia (DRG) transmit sensory information such as pain, itch, touch, pressure and bodily position to the central nervous system. The activity of sensory neurons is regulated by non-neuronal cell types in the DRG, including satellite glial cells (SGCs), immune cells and fibroblasts. Dysregulated gene expression in DRG cells contributes to sensory nervous system disorders such as chronic pain. Understanding the genetic underpinnings of these conditions requires dissecting transcriptional regulation in human tissue. In this study, we profiled transcriptomic and chromatin accessibility landscapes from postmortem human DRG (hDRG) samples at single-nucleus level. We demonstrate that sequencing depth significantly impacts downstream analysis, with deeper sequencing yielding more detected cells and features, improved data integration, refined clustering and annotation, and more accurate scientific interpretations. We identified nine major cell types, defined their molecular signatures, and mapped cis-regulatory landscapes. Integration of gene expression with chromatin accessibility enabled peak-gene association and transcriptional network analyses, revealing transcription factors, their target genes, regulatory elements and potential partners that cooperatively drive cell-type-specific gene expression programs. This integrative approach identified cell types, genes, and cis-regulatory regions potentially driving pain conditions. Our unbiased genome-wide analysis not only recovered known pain-related genes but also highlighted novel candidate genes and regulatory regions implicated in pain mechanisms. Importantly, our results demonstrate that non-neuronal cells, including endothelial cells, fibroblasts, macrophages, and SGCs, play critical roles in pain pathogenesis and should be investigated as therapeutic targets. One Sentence SummaryOur work provides a comprehensive single-nucleus multi-omic atlas of human dorsal root ganglia, uncovering novel cell-type-specific regulatory mechanisms and candidate therapeutic targets for pain, thereby directly advancing translational insights into human sensory disorders and chronic pain pathogenesis.

neuroscience↗

A multi-omic atlas of human autonomic and sensory ganglia implicates cell types in peripheral neuropathies

The human peripheral nervous system (PNS) consists of many ganglia including sympathetic ganglia (SG) and dorsal root ganglia (DRG). These ganglia house the cell bodies of diverse PNS neurons that transmit autonomic and sensory signals, as well as a much larger number of non-neuronal cells. However, the molecular and cellular diversity of these human PNS cell types and their implications in human disease remain elusive. By generating an integrated single-cell multi-omic atlas of human SG and DRG, we provide comprehensive transcriptional and epigenomic landscapes of various cell types in these peripheral ganglia. While the major cell types and their cell-type-specific transcriptional and epigenomic features are similar between human SG and DRG, we identify key differences between SG and DRG cell types. These differences highlight the distinct molecular and cellular mechanisms underlying their specific functions. We also profiled key genomic regulatory networks that govern cell-type-specific gene expression in these peripheral ganglia. Moreover, by mapping the expression and chromatin accessibility of disease-associated genes in human SG and DRG, we identify cell types that may underlie various peripheral neuropathies. This atlas serves as a valuable resource for understanding the intricate cell-type-specific molecules and interactions in the human PNS and their implications in human health and diseases.

neuroscience↗