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Biology subjects

Singh, P.

Publications and source records attributed to Singh, P..

At least 19 recordsLinked to original sources

Imputing relative abundances via Gaussian random fields

Longitudinal studies with human participants frequently face the challenge of missing samples, which lead to incomplete trajectories of participant biomarkers and a reduction in data size. Existing approaches focus on single-variable imputation or model features one at a time, while neural networks typically require a large number of parameters and substantial training data. To address these challenges, we present an explainable model for multiple imputation of compositional samples based on a Gaussian random field (GRF). The model is specifically designed for longitudinal data: it incorporates participant-specific information and accounts for both temporal dependence across repeated observations and within-sample correlations, while using a moderate number of parameters. By coupling the GRF with sparse activation functions, the framework outputs sum-constrained compositions and explicitly models zero inflation. Under the Aitchison and Jensen--Shannon distances, our framework achieves imputation accuracy comparable to that of an observation-carrying baseline, while overcoming its static nature by modeling temporal trajectories and multivariate correlations. The resulting multiple imputations allow us to reconstruct complete datasets for downstream analysis and uncertainty quantification. We demonstrate the practical utility of our framework on gut microbiome datasets, restoring incomplete time series and achieving a balanced accuracy of 0.72 in predicting food allergy in infants (sensitivity 0.8) and of 0.64 in forecasting postpartum depression.

bioinformatics↗

Simple soil eDNA sampling reveals terrestrial mammal diversity in the Central Kalahari and highlights waterholes as sampling hotspots

Remote and vast ecosystems present a challenge for biodiversity monitoring. The Central Kalahari Game Reserve in Botswana is one such ecosystem spanning 52,800 km2, a mosaic of fossilized rivers, clay pans, semi-arid sand dunes, scrub, and woodland. It is one of the last remaining untouched wildernesses in the world and hosts unique biodiversity, including populations of threatened mammals. Due to its extreme arid environment, size, and inaccessibility, surveying biodiversity across the reserve is both logistically and technically difficult. Here we demonstrate that a low-effort approach of sampling soil across the reserve is an efficient way to rapidly assess the terrestrial mammal biodiversity using eDNA metabarcoding. We found that eDNA from soil near waterholes performed better at capturing local biodiversity than soil from roads. eDNA metabarcoding detected 38 mammal species, 15 bird species and two amphibians and a simultaneously conducted camera trapping study detected 29 mammal species. eDNA was particularly powerful for detecting small-bodied taxa, including murids, mongooses and bats, which were largely missed by camera traps, while camera trapping was aimed at and performed well for detecting felids. Mammal species that had a higher activity nearby eDNA sampling points (i.e., a higher number of camera trapping recordings nearby) had a higher probability of being detected via eDNA. We propose that simple soil collection for eDNA metabarcoding, particularly close to animal congregation points such as savanna waterholes, offers a practical, scalable and cost-effective strategy for DNA-based biomonitoring in large, remote areas such those in Southern Africa.

ecology↗

Reply to: Structural polymorphism of amyloid fibrils in ATTR amyloidosis revealed by cryo-electron microscopy

In the paper by Nguyen et al, published in Nature Communications in 2024, we describe structural polymorphism in fibrils extracted from patients with ATTR amyloidosis. ATTR amyloidosis is characterized by deposition of transthyretin amyloid fibrils across multiple organs, resulting in marked phenotypic variability. Previous studies on ATTR suggest its phenotypic variability may be linked to differences in fibril structure, as demonstrated in amyloid diseases such as AA amyloidosis, tauopathies, or synucleinopathies. Thus far, most ATTR fibrils adopt a strikingly similar fold across patients and affected organs. The most notable exception is described in the study by Nguyen et al., where we report the identification of two co-existing polymorphs in cardiac fibrils from three patients with the ATTRv-I84S variant. These cardiac polymorphs differ in conformation of the segment Gly57 to Gly67, termed the gate, which regulates access to a central polar channel within the fibril core. In one polymorph, the gate adopts an open conformation, exposing the channel, whereas in the other it is closed, shielding it. Whether these polymorphs extend beyond cardiac tissue remains unknown. Here, we address this question by extending our analysis to additional organs from a fourth patient with ATTRv-I84S amyloidosis.

biophysics↗

Patient-Derived Melanoma Organoids Preserve Tumor-Immune Heterogeneity and Reveal Context-Dependent Responses to Immune Checkpoint Blockade

Abstract Background: Experimental models that retain endogenous tumor immune complexity are needed to investigate heterogeneous responses to immune checkpoint inhibitors (ICIs) in melanoma. We established patient-derived melanoma organoids (PDMOs) to examine retention of parental tumor cellular components and investigate patient-specific and context-dependent responses to checkpoint blockade. Methods: Fresh melanoma specimens (n=50), including primary tumors, neoadjuvant-treated tumors and tumor-infiltrating lymphocyte (TIL) associated samples were processed for Matrigel-embedded culture of patient-derived melanoma organoids (PDMOs). Characterization and functional studies were performed in subsets of established cultures. Immunofluorescence assessed tumor, stromal, immune, and checkpoint-marker expression in PDMOs and matched parental tissues. NGFR expression was evaluated in relation to organoid establishment and growth. Responses to anti PD1 alone or combined with anti-LAG3 or anti-CTLA4 therapies were evaluated using viability and morphological analyses, with selected models undergoing immune phenotyping and multiplex cytokine profiling. Available clinical outcomes were used for exploratory comparison. PDMOs and matched two-dimensional cultures were evaluated under 21% and 5% oxygen conditions. Results: PDMOs were established from 30 of 50 specimens (60%) and retained melanoma, stromal, lymphoid, and myeloid components identified in matched parental tissues during early culture, together with immune checkpoint expression. Higher NGFR (CD271) expression was associated with greater organoid-forming capacity in an exploratory subset. Responses to checkpoint blockade varied across patient-derived models and regimens, with combination blockade not uniformly producing lower viability than PD1 monotherapy. Ex vivo responses showed parallels with available clinical outcomes, with discordant cases also observed. In three selected PDMOs, divergent PD-1 responses were accompanied by differences in T-cell and myeloid representation, MHCII and AXL expression, and inflammatory versus immunoregulatory cytokine profiles. Under normoxic conditions, PDMOs and matched two-dimensional cultures exhibited divergent treatment sensitivities, including clinically responding cases in which sensitivity was retained in PDMOs but attenuated in monolayers. Changing oxygen tension further altered checkpoint sensitivity within individual PDMOs, with the direction and magnitude of the observed shifts varying by model and regimen. Conclusions: Early passage PDMOs retain key endogenous tumor, immune, and stromal components and support functional investigation of patient-derived melanoma biology. Their heterogenous responses across checkpoint regimens and oxygen conditions provide a platform for investigating how tumor-immune composition and environmental context influence treatment sensitivity.

immunology↗

AGC kinase homology requires and enables co-targeting for CNS regeneration

Axon regrowth in the central nervous system (CNS) is constrained by robust regulatory networks. Here we show that optimal neurite outgrowth in rodent and human CNS neurons is achieved by co-inhibition of kinases across four closely related clades within the protein kinase A, G, and C (AGC) family. The kinases derive from ancestral regulators of cytoskeletal dynamics, resource allocation, and polarized cell growth. Their shared domain architecture makes polypharmacology (co-engagement by a single small molecule) feasible. Phenotype-guided optimization of a tool compound with established efficacy in mouse spinal cord injury models yielded TMP-316, a drug candidate engaging these AGC kinases with selectivity against the broader kinome. A single intrathecal dose of TMP-316 produced sustained motor recovery in a rat cervical hemicontusion model. These findings reveal conditions under which polypharmacology is simultaneously required and enabled by shared evolutionary origins, illuminating a therapeutic discovery principle for pathologies governed by functionally overlapping targets.

neuroscience↗

HuMMANet: A Harmonized Cross-Study Resource for Integrative Analysis of Human Gut Microbiome Metabolome Associations

Deciphering gut microbiome to host metabolome interaction is critical for understanding how microbial communities generate bioactive signals that shape host physiology and disease. Progress, however, has been hindered by inconsistent metabolite annotations, poor interoperability across studies, and the absence of integrated resources placing microbiome-derived metabolites within their functional, microbial, physiological, and clinical context. Here we present HuMMANet (Human Microbiome Metabolome Annotation Network), a harmonized resource integrating 46 paired gut microbiome metabolome studies (59 study-units; 14,405 samples; 13 disease categories plus a healthy/control reference category) with a scalable metabolite-harmonization framework. HuMMANet resolves heterogeneous annotations through a multi-stage workflow spanning RefMet, HMDB, PubChem, Metabolomics Workbench, SMPDB, MiMeDB 2.0, GNPS/microbeMASST, DrugBank, and DrugCentral, yielding a reference atlas of 54,914 unique metabolites, annotated with standardized chemical identifiers, biochemical pathways, microbial producer associations, physiological distributions, disease links, and structural relationships to approved therapeutics, a unified reference framework for microbiome metabolome research. Applying HuMMANet to a multi-cohort integration of adult serum and fecal metabolomes, we identified 519 serum and 322 fecal metabolites reproducibly associated with gut microbial community composition (PERMANOVA, P < 0.05 in at least 50% of studies in which detected), enriched for specific biomolecular classes and pathways. Cross-referencing these against Health Associated Core Keystone (HACK) taxa revealed 58 serum and 25 fecal metabolites (HACK positive) whose taxon-level associations tracked positively with the taxon specific HACK indices. These reproducible metabolomic signatures of microbiome health included indole3propionic acid, a gut barrier-protective microbial tryptophan metabolite, and 3phenylpropionate. Drug similarity annotation within HuMMANet linked 16 of this serum and 13 fecal HACK positive metabolites to therapeutics used in neurological, inflammatory, and vascular disease. Conversely, 38 serum and 65 fecal metabolites, including imidazole propionate and long-chain acylcarnitines such as ACar 18:0, showed HACK negative signatures previously associated with dysbiosis-linked disease. GNPS/microbeMASST and MiMeDB 2.0 annotations further traced subsets of these metabolites to putative bacterial producers. HuMMANet thus provides a standardized framework for reproducible microbiome metabolome integration, enabling cross study discovery and translational prioritization of conserved microbiome derived metabolic signatures across human populations and disease states.

bioinformatics↗

Imaging guided single-cell multiomics unveils shared autoreactive CD4+ T-cell responses in blood, locoregional lymph node and affected tissues of patients with systemic autoimmunity

Systemic autoimmune connective tissue diseases (CTDs) are characterized by anti-nuclear antibodies, shared HLA-associated genetic risk, and frequent disease overlap, suggesting a central role for CD4+ T cells in pathogenesis. However, defining disease-driving CD4+ T-cell responses remains challenging due to their localization within lymphoid and affected tissues and the lack of approaches linking these responses to circulating counterparts. We combined [18F]-labeled thymidine PET/CT-guided tissue sampling, ex vivo antigen stimulation, and single-cell multiomics to characterize CD4+ T-cell responses in blood, PET-avid locoregional lymph nodes (LNs), and disease-affected tissues from patients with the immunologically distinct CTDs systemic sclerosis and Sjogren's disease. PET-avid LNs from both diseases exhibited enhanced adaptive immune activity and contained an expanded population of interferon-stimulated gene (ISG)-expressing TRAIL+ CD4+ T cells. In Sjogren's disease, active LNs and affected tissues harbored diverse effector CD4+ T-cell populations, including follicular and peripheral helper T cells and Th2/Th17 cells. In contrast, systemic sclerosis tissues lacked effector CD4+ T cells, while active LNs were enriched for naive, regulatory, and TRAIL+ ISG CD4+ T cells. Antigen stimulation of peripheral blood mononuclear cells enriched for expanded effector CD4+ T-cell populations that shared activation profiles and clonal relationships with cells in LNs and affected tissues, many representing autoreactive antigen-specific T cells. TRAIL+ CD4+ T cells suppressed effector T-cell differentiation, autoreactive plasma cell generation, and autoantibody production in vitro, identifying a previously unrecognized immunoregulatory population. Together, this workflow enables comprehensive characterization of pathogenic and regulatory CD4+ T-cell responses across CTDs.

immunology↗

MifRix: An Integrated microbiome framework for predicting generic and disease-specific risks investigating inter-disease diagnostic cross-talks and intra-disease signature variability

The human gut microbiome is increasingly recognized as a diagnostic indicator across diverse diseases, yet unified frameworks integrating taxonomic and functional features for multi-disease risk assessment, while capturing disease-specific variability in microbiome-alteration signatures, remain limited. Here we present MifRix (Microbiome-inferred Risk-scores with explainability), a two-step ensemble machine-learning framework integrating microbial composition with composition-derived functional signatures to predict generic and disease-specific risk scores across 10 major diseases, coupled with profiling of risk-explainable microbiome features. MifRix was trained using 38,054 gut microbiomes spanning 150 cohorts and 48 nationalities, leveraging taxa abundance and taxa-inferred functional profiles derived from 57,743 functional features mapped across 4,814 species-level taxa. On unseen validation datasets (4,649 microbiomes, 32 cohorts), MifRix outperformed established microbiome health metrics, with disease-specific risk scores achieving strong discrimination (AUC: 0.92-0.99). The explainability module revealed shared microbial signatures across disease pairs, organizing all ten diseases along a continuous gastrointestinal-to-neurological risk gradient, driven not by whole-disease microbiome alteration signatures but by specific, reproducible sub-signatures within each disease. Applied across independent cohorts, MifRix scores further identified population subgroups and individuals at elevated risk for related diseases, flagged precursor/pre-disease states, and tracked therapy-associated response, establishing an interpretable framework for microbiome-based precision diagnostics and disease-risk stratification.

microbiology↗

Oncogenic transformation proceeds through a transient state of cellular plasticity constrained by lineage-specific barriers.

Normal tissues frequently harbor oncogenic mutations without progressing to cancer, but the cellular basis of this resistance remains poorly defined. We asked whether transformation requires selection of a rare, permissive state within normal-like cell populations. Dermal fibroblasts and mammary epithelial cells responded uniformly to combined HRAS-G12V expression and p53 disruption. Cellular barcoding revealed no loss or enrichment of clones during morphological transformation, arguing against clonal selection as the primary driver of neoplastic reprogramming. Instead, all transduced cells underwent an early, shared transcriptional transition characterized by loss of differentiation markers, induction of RAS-associated and inflammatory programs, activation of alternative-lineage signatures, increased single-cell entropy, and chromatin decondensation. These changes were transient: entropy and chromatin accessibility subsequently declined, and some cells moved toward the control transcriptional state, whereas others stabilized in altered states. The two lineages followed distinct trajectories. Fibroblasts showed greater initial transcriptional plasticity but subsequently reverted more strongly toward the normal state, whereas epithelial cells changed more gradually and continued to diverge from it, suggesting a stronger barrier to transformation in the mesenchymal lineage. Thus, oncogenic perturbation initiated reprogramming throughout the population but did not uniformly produce a stable transformed state. Together, these findings support a model in which normal-like cells tolerate oncogenic mutations not because most cells fail to respond, but because a p53-independent, cell-intrinsic barrier limits the stabilization of malignant transformation following a transient period of heightened plasticity. This framework may facilitate the identification of mechanisms that constrain tumor initiation.. Significance StatementLoss of TP53 and activation of oncogenic RAS are common drivers of human cancer and frequently coexist in the same tumor. Nevertheless, these alterations often fail to induce malignant transformation. To investigate why, we introduced both alterations into fibroblasts and epithelial cells and tracked their responses using clonal barcoding, single-cell transcriptomics, and phenotypic assays. Initially, cells uniformly reprogrammed their transcriptional and chromatin states without detectable clonal selection. They then diverged: some reacquired transcriptional profiles resembling those of the original normal cells, whereas others became transformed. Thus, normal cells resist transformation through an intrinsic barrier that operates despite p53 disruption. Because this transition is transient and reversible, it provides a tractable window for studying the earliest stages of cancer development.

cancer biology↗

Identification of pre-existing ubiquitous neoantigen-reactive tumor-infiltrating T-cells in a patient with metastatic pancreatic neuroendocrine tumor

BackgroundMutation-derived neoantigens, typically identified in primary tumors, are emerging therapeutic targets for personalized cancer vaccines and adoptive T-cell therapies. However, clinical efficacy of neoantigen-directed therapies in patients with metastatic disease remains limited, partly due to inter-site genetic heterogeneity. We investigated whether ubiquitous neoantigens-derived from mutations shared across all tumor sites-could provide more effective, durable targets, particularly in patients undergoing resection of metastatic lesions. MethodsWhole-exome and RNA sequencing were performed on 14 tumor samples (primary and 13 synchronous nodal metastases) from a treatment-naive patient with pancreatic neuroendocrine tumor (PNET). Ubiquitous mutations were identified bioinformatically, and their immunogenicity assessed using in-vitro stimulation of autologous peripheral blood mononuclear cells followed by IFN-{gamma} ELISpot assay. Neoantigen-specific T-cell clonotypes were further identified by HLA-tetramer staining and single-cell RNA/TCR sequencing. Neoantigen-reactive clonotypes identified in peripheral blood were tracked across multiple metastatic sites using bulk TCR{beta} repertoire sequencing. ResultsAmong 1,195 non-synonymous mutations detected, eight were shared across all 14 tumor sites. Of these, one encoded a neoantigen that elicited a reproducible IFN-{gamma} ELISpot response in peripheral blood, confirming its immunogenicity. Further, we identified the corresponding neoantigen-reactive TCR clonotypes in blood. Comparison with bulk TCR{beta} repertoires from eight metastatic sites showed that these clonotypes were present in every site analyzed, with evidence of local clonal expansion. ConclusionThis study provides direct evidence that a single ubiquitous mutation-derived neoantigen can generate systemic T-cell responses and clonotype expansion across multiple metastatic sites in a TMB-low, TIL-low tumor. Our findings support incorporating mutation-sharing status across metastases as a key criterion for neoantigen selection in cancer vaccines and adoptive T-cell therapies. This approach could inform the design of neoantigen-directed immunotherapies in metastatic PNET and potentially other metastatic solid tumors. What is already known on this topicNeoantigen-directed therapies, such as personalized cancer vaccines or adoptive T-cell transfer, can induce anti-tumor responses but have shown limited success in metastatic disease. One major barrier is genetic heterogeneity between tumor sites, suggesting that targeting ubiquitous mutations-those shared across all tumor sites-may improve the efficacy of such therapies. What this study addsIn one patient with metastatic pancreatic neuroendocrine tumor involving 13 lymph nodes, we identified eight ubiquitous mutations, one of which generated a detectable neoantigen-specific T-cell response in blood. The corresponding T-cell clonotypes were found across all metastatic sites analyzed and showed evidence of clonal expansion, providing direct evidence of systemic and local recognition of a shared neoantigen in a TMB-low/TIL-low cancer. How this study might affect research, practice or policyThese findings support incorporating mutation sharing across metastases as a key criterion in neoantigen selection for cancer vaccines and adoptive T-cell therapies. This strategy could enhance the relevance and durability of neoantigen- directed approaches in patients with metastatic disease.

immunology↗

An Atomistic Description of Heterotypic Lipid Exchange by Sec14-like Phosphatidylinositol Transfer Proteins

Lipid transfer proteins (LTPs) are core regulators of the membrane dynamics, lipid signaling and intracellular communication networks that connect every organelle in the eukaryotic cell. ATP-independent lipid exchange reactions are a hallmark activity of these proteins. These remarkable reactions are essential for the important biological functions of LTPs but how lipid exchange is executed is not at all understood. Herein, we focus on phosphatidylinositol transfer proteins (PITPs) of the highly conserved and highly expanded Sec14/CRAL-Trio-like protein superfamily that potentiate phosphatidylinositol-4-phosphate (PtdIns4P) signaling in eukaryotic cells. Using an integrated structural approach, we describe in atomistic detail the lipid exchange reaction of Sec14-like PITPs. The molecular concepts we identify not only yield insights into how these PITPs integrate metabolic activity with PtdIns4P signaling in cells but also provide a framework for interpreting the functional mechanisms of other LTPs of the Sec14/CRAL-Trio superfamily.

biophysics↗

A large-scale crowd-sourced annotated acoustic dataset of Indian fauna

Global rates of biodiversity loss warrant conservation action and monitoring at large geographic scales. Conservation technologies such as acoustic monitoring in conjunction with deep learning now enable us to monitor wildlife simultaneously across space and time. However, for a significant proportion of biodiversity in tropical regions, we cannot yet rely on automated recognition approaches because we lack acoustic templates to robustly train deep learning algorithms. In this paper, we relied on a novel participatory approach, enlisting researchers, conservation practitioners, and nature enthusiasts to create a unique crowd-sourced, open-access dataset of acoustic annotations across taxonomic groups for biodiversity in India. Our dataset comprises 3311 minutes of strongly labelled data (bounding boxes or annotations for a species vocalization) and 2504 minutes of weakly labelled data (indicating the presence of a species within an audio file but lacking bounding boxes) for 518 species across India, spanning 25 of 36 states and union territories. We present metadata and code for data processing and highlight the strengths of a participatory approach to biodiversity monitoring.

ecology↗

Plant Functional Type Composition, Rather Than Species Diversity, Shapes Soil Microbial Functional Diversity and Redundancy

Cover crops (CCs) are widely used to improve soil health, but their species-specific effects on microbial functional diversity and redundancy remain poorly understood. We evaluated how monocultures of field pea (Pisum sativum), forage radish (Raphanus sativus), and cereal rye (Secale cereale), as well as a three-species mixture (3spp) and a five-species mixture (5spp), influence rhizosphere bacterial and fungal communities in a field experiment. Amplicon sequencing and predictive functional profiling were used to assess microbial diversity, composition, functional potential, and functional redundancy (FR). Microbial alpha diversity showed little change across treatments, but community composition and predicted functional profiles were strongly influenced by CC identity. The 3spp rhizobiome showed the highest bacterial FR, broad metabolic capacity, and greater network connectivity, suggesting increased functional stability of all treatments. In contrast, the 5spp rhizobiome showed reduced bacterial FR and more widespread functional depletion, likely due to imbalances in plant functional types. Monocultures showed species-specific patterns, with rye supporting functionally efficient communities and radish promoting more competitive interactions and reduced functional diversity. Fungal communities responded differently from bacterial communities. While fungal taxonomic shifts were limited, they showed stronger compositional differences among treatments and more stable functional profiles. Notably, fungal FR was highest in the 5spp rhizobiome. Overall, these findings highlight that balanced plant functional composition, rather than greater species richness alone, is important for shaping rhizosphere microbial function and redundancy. Balanced mixtures promoted higher bacterial redundancy, while more diverse but functionally imbalanced mixtures did not consistently enhance microbial function. ImportanceThe benefits of using CCs in agricultural settings are mostly influenced by microbial communities, including nutrient cycling, organic matter turnover, and resilience to disturbance. However, CC mixtures are often promoted on the assumption that more species provide greater soil benefits. This work challenges that assumption by showing that the functional identity and balance of CC species may matter more than the number of species alone. This distinction is important for farmers, land managers, and researchers because CC seed mixes can be costly, and poorly balanced mixtures may fail to deliver intended microbial benefits. Understanding which plant combinations support functionally stable microbial communities can improve CC recommendations and help design agricultural systems that maintain soil processes under changing environmental conditions.

microbiology↗

Proprotein convertase subtilisin kexin type 9 (PCSK9) inhibition attenuates abdominal aortic aneurysm formation via enhanced macrophage-dependent efferocytosis

Abdominal aortic aneurysms (AAAs) occur predominantly in the elderly population and currently there is no effective pharmacological therapy for mitigating AAA growth and preventing impending rupture. Proprotein subtilisin kexin type 9 (PCSK9) gene has been identified as a specific risk-locus for AAA development. However, the mechanistic and clinical role of PCSK9-mediated signaling in AAAs has not been delineated. We demonstrate that treatment with PCSK9 inhibitors, such as Evolocumab, mitigates vascular inflammation and remodeling, resulting in attenuated aneurysm growth in clinical datasets as well as experimental models of AAA and aortic rupture. Mechanistically, Evolocumab immunomodulates macrophage reprogramming to enhance clearance of apoptotic smooth muscle cells via MerTK-dependent efferocytosis that ameliorates aortic inflammation and vascular remodeling. Furthermore, Evolocumab increases the expression of oxidized phosphatidylserine species and decreases expression of lysophospholipids, succinate, and glycolytic intermediates within the aortic wall compared to untreated controls, further enhancing the pro-resolving functions of macrophages. Collectively, our data demonstrates the ability of PCSK9 inhibition to regulate macrophage-specific efferocytosis that limits AAA progression and prevents aortic rupture.

immunology↗

Cereal grafting in rice and pearl millet preserves photosynthetic performance and stomatal dynamics, establishing a platform for root-shoot communication studies

O_LICereal grafting using embryonic tissues has recently become technically feasible; however, the physiological consequences of cereal grafting remain uncharacterized. C_LIO_LIWe systematically evaluate photosynthetic performance and stomatal dynamics across different graft combinations in two photosynthetically distinct species, rice (C3) and pearl millet (C4). We first assessed steady-state photosynthetic performance and dynamic stomatal responses in five-week-old rice and pearl millet grafts grown under a saturated water regime, to establish whether cereal grafting alters physiology at early stages. Next, we assessed same traits at the onset of optimal water regime, and after five days to determine whether any graft-induced effects on photosynthesis or growth persisted over time. C_LIO_LIWe observed that across contrasting water regimes and at different plant developmental stages, cereal grafting did not alter growth, photosynthesis or stomatal kinetics in either species, while revealing modest early stage C4-specific adjustments in stomatal dynamics without affecting photosynthetic capacity or biochemical parameters. C_LIO_LIWe demonstrate that cereal grafting does not alter core physiological traits in rice or pearl millet and can be deployed without long-term impact on photosynthesis. These findings establish cereal grafting as a tractable platform for mechanistic dissection of root-shoot signaling and trait combination across different C3 and C4 cereals. C_LI

Plant Biology↗

Microbiota-derived indole limits Campylobacter jejuni colonization by inhibiting respiration and metabolism

How the microaerophilic Campylobacter jejuni grows in the inflamed gut is poorly understood, hampering our ability to identify novel targets for controlling infections by this pathogen. Our prior ferret study indicated that C. jejuni growth during infection is driven by intestinal inflammation. Without manipulating microbiota or introducing pro-inflammatory genetic lesions (or both), conventional mice are naturally resistant to C. jejuni colonization and infection. To test the impact of inflammation per se on C. jejuni infection, we induced transient intestinal inflammation in mice using short-term dextran sodium sulfate (DSS) treatment. The resulting colitis disrupted colonization resistance, enabling rapid C. jejuni growth in the murine colon within three days of infection, accompanied by exacerbated intestinal inflammation. DSS-induced colitis led to enrichment of mucin-degrading bacteria and depletion of taxa producing short-chain fatty acids and indole; metabolomic profiling confirmed a marked reduction in colonic indole levels in both DSS-treated and infected mice. At physiological concentrations, indole inhibited C. jejuni growth in vitro and resulted in reduced transcript levels from key energy-generating pathways, including nitrate respiration (napA), aerobic respiration (ccoN), lactate utilization (lctP), and the acetate switch (ackA/ptaA); consistent with this, mutations in these pathways led to fitness defects in DSS-treated mice, highlighting their importance for C. jejuni colonization in the inflamed gut. Moreover, treatment with indole or the indole-producing probiotic Escherichia coli Nissle 1917 significantly reduced C. jejuni colonization in vivo. Our findings show how C. jejuni establishes and grows in the inflamed gut and demonstrate that microbiota-derived metabolites play a key role in regulating C. jejuni pathogenicity. SignificanceThe mechanisms by which the microaerophilic pathogen C. jejuni proliferates in the inflamed intestine remain poorly understood. Using a DSS-induced colitis mouse model, we identify host physiological changes, microbiota alterations, and metabolic factors that promote C. jejuni colonization during intestinal inflammation. These findings provide mechanistic insight into why Campylobacter species are frequently detected in patients with Inflammatory Bowel Disease (IBD) and how infection can exacerbate intestinal inflammation and worsen disease symptoms. We further identify the microbiota-derived metabolite indole as a potent inhibitor of C. jejuni colonization, highlighting a potential metabolite-based therapeutic strategy to combat emerging multidrug-resistant C. jejuni infections.

microbiology↗

Synovial transcriptional clusters link cartilage degeneration to cell-type-specific gene expression in knee osteoarthritis

ObjectivesTo identify synovial transcriptional clusters in human knee osteoarthritis (OA) and determine how these relate to synovial histologic features, cell-type-associated gene expression, and cartilage degeneration severity. MethodsBulk RNA sequencing (RNA-seq) of synovial tissue from n = 135 patients with knee OA was analyzed using consensus clustering. Clusters were compared by clinical and histologic features, including cartilage degeneration severity (OARSI score). Single-cell RNA-seq (n = 18) and spatial transcriptomics were used to relate cartilage degeneration-associated gene expression patterns to synovial cell populations. ResultsFour synovial transcriptional clusters that differed in synovial histologic features and cartilage degeneration severity were identified. Greater cartilage degeneration was associated with enrichment of lining fibroblast- and inflammatory myeloid-associated gene expression, whereas lesser cartilage degeneration was associated with enrichment of sublining fibroblast, endothelial, mural cell, and adipocyte-associated gene expression. ConclusionsHuman knee OA synovium segregates into transcriptional clusters associated with cartilage degeneration severity. Synovial transcriptional heterogeneity corresponds to cell-type-associated gene expression. Key messagesO_ST_ABSWhat is already known on this topicC_ST_ABSO_LIOsteoarthritis synovium exhibits marked histologic and molecular heterogeneity. C_LIO_LISynovial inflammation detected by MRI correlates with cartilage degeneration and predicts progressive cartilage loss in knee OA. C_LIO_LIPrior transcriptomic studies have identified molecular subsets of OA synovium, but their relationship to cartilage degeneration severity remains unclear. C_LI What this study addsO_LIOA synovium segregates into four transcriptional clusters: Sublining (C1), Lymphomyeloid (C2), Myeloid (C3), and Major trauma (C4). C_LIO_LIGreater cartilage degeneration is associated with enrichment of inflammatory myeloid and lining fibroblast gene expression, whereas lesser degeneration is associated with enrichment of adipocyte, sublining fibroblast, endothelial, and mural cell-associated gene expression. C_LI How this study might affect research, practice or policyO_LIProvides a framework for a clinically relevant biological stratification of OA patients based on synovial molecular features. C_LIO_LIInforms future efforts to link synovial biology with OA prognosis, cartilage degeneration, treatment allocation, and development of targeted therapeutic strategies. C_LI

genomics↗

Biodiversity dynamics with complex genotype-to-phenotype architecture in multilayer networks

2The genotype-to-phenotype architecture (GPA), defined by complex interactions such as pleiotropy, epistasis, and regulatory control, is a fundamental yet often overlooked driver of biodiversity dynamics. While empirical evidence suggests that traits mediating species interactions (biotic) and environmental responses (abiotic) are frequently correlated, most eco-evolutionary theories treat these traits as independent, leaving a gap in our understanding of how genomic architecture influences community-level outcomes. In this study, we contrast two distinct GPAs, modular (independent trait evolution) and correlated (integrated trait evolution), within a spatially explicit multilayer network framework. We evaluate their impact on biodiversity across varying regimes of selection, migration, and biotic and environmental filtering. Our results reveal a hierarchy of drivers: selection strength dictates the absolute magnitude of the architectural effect, while migration and context-dependent biotic and abiotic effects determine which architecture yields a diversity advantage. Correlated GPAs enhance species coexistence and diversity in low-migration landscapes characterized by strong selection and moderate, balanced biotic and abiotic pressures. In these contexts, trait integration serves as a buffer against selective noise. Conversely, modular GPAs support higher diversity under high migration and strong biotic interactions, where the decoupling of trait modules provides the adaptive flexibility necessary to navigate spatially conflicting selective pressures. Our findings demonstrate that genomic architecture acts as a critical filter for environmental perturbations. Integrating complex GPAs into multispecies models is essential for quantifying the co-evolutionary feedbacks among traits, population adaptation, and species persistence. Our framework provides a path for predicting how biodiversity emerges and persists across biological scales, from genomics to communities and food webs, under the accelerating pressures of global change. 1 ConclusionsO_LIWe integrate trait architecture to spatial biodiversity to show biodiversity patterns are not merely products of ecological interactions, but are fundamentally constrained by Genotype-to-Phenotype Architecture (GPA). By linking GPA to biodiversity we show the interplay between the complexity of an organism and community structure in determining diversity patterns. C_LIO_LIThe hierarchy of Eco-Evolutionary Drivers: We establish a new conceptual hierarchy where selection strength acts as the fundamental governor of architectural impact, while the specific architecture predicting higher diversity (Correlational vs. Modular) is dictated by the interplay of migration scales and context-dependent biotic and abiotic dynamics. C_LIO_LISelection-Migration contingency for coexistence: We provide a new hypothesis for species coexistence: Correlational selection serves as a stabilizing force under dispersal limitation, whereas Modular trait architecture provides the adaptive flexibility to maintain diversity in high-migration, spatially heterogeneous landscapes. C_LIO_LIAdaptive decoupling as a diversity engine: We propose that trait modularity functions as a "buffer" against extinction by decoupling phenotypic responses. This allows populations to navigate conflicting selective pressures, effectively facilitating evolutionary rescue in complex biotic environments. C_LIO_LIMethodological framework for empirical inference: To bridge the gap between theory and data, we provide a novel likelihood-based framework. This enables researchers to infer latent trait architectures from population genomic samplings, turning GPA from a theoretical construct into a measurable sampling variable in natural populations. C_LIO_LIWe define a new roadmap for the next generation of eco-evolutionary modeling. By identifying the gaps between existing simulation engines, we provide a conceptual "blueprint" for a digital ecosystem that fully integrates complex genetic architecture with global bio-diversity dynamics. C_LI

evolutionary biology↗