Biology subjects

Zheng, J.

Publications and source records attributed to Zheng, J..

At least 19 recordsLinked to original sources

bioRxiv2026

ClonoDeconv: reference-based α - β paired recurrent clonotype proportion estimation from bulk-TCR sequencing

T cell receptor (TCR) sequencing is crucial when studying antigen-specific T cell responses and clonal dynamics. A key objective of TCR repertoire analysis is to identify recurring clonotypes across timepoints or different disease states, providing insight into the evolution of immune response. Bulk-TCR sequencing enables high-throughput profiling of TCR and {beta} sequences but lacks pairing information, while single cell TCR sequencing (scTCR-seq) retains pairing information but is limited by sensitivity, throughput, and costs. We present ClonoDeconv, a computational framework to deconvolute paired TCR {beta} ; clonotype frequency from bulk TCR sequencing experiments using a paired scTCR-seq reference. ClonoDeconv robustly estimated shared clonotype abundances from 5' RACE bulk-TCR datasets using a scTCR-seq reference from the same donor. Applied to longitudinal samples, ClonoDeonv successfully captured the expansion and contraction of rare virus-specific clones across timepoints. Taken together, ClonoDeconv provides a cost-effective and scalable approach for studying the longitudinal T-cell clonal dynamics.

bioinformatics↗

bioRxiv2026

Pyruvate partitioning defines an immunosuppressive metabolic niche in colorectal cancer

Purpose. Colorectal cancer (CRC) reprograms glucose metabolism, diverting pyruvate from mitochondrial oxidation toward lactate export. How this partitioning decision is organized across cell types and tissue space, and how it relates to immune programmes, remains unresolved. Methods. We defined a Pyruvate Partitioning Index (PPI) as the difference between an oxidative arm (MPC1, MPC2, PDHA1, PDHB, DLAT, DLD, PDHX) and an export arm (LDHA, LDHB, SLC16A1, SLC16A3, SLC16A7), and computed it across TCGA COADREAD (n = 592), GSE39582 (n = 579), two single-cell atlases (370,115 and 63,689 cells), 13 Visium sections and an exploratory urothelial immunotherapy cohort (IMvigor210, n = 298). Results. PPI was lowest in microsatellite-unstable (MSI) tumours (Mann-Whitney p = 3.8 x 10^-9) and inversely associated with overall survival (log-rank p = 1.2 x 10^-4). Epithelial cells were the most oxidative compartment (median +0.058) and myeloid cells the most export-prone (-0.171; Kruskal p [~] 0), reproduced independently; MMRd epithelium was intrinsically export-shifted (-0.012 vs +0.076 vs +0.192 for MMRd, MMRp and normal). Spatially, PPI was non-random (Moran's I significant in 12/13 sections), with low-PPI spots enriched for SPP1+ macrophages, myofibroblasts and plasma cells, but not CD8+ T-cell factors. Low-PPI tumours showed coordinated increases in immune-checkpoint, myeloid and Treg programmes (checkpoint rho = -0.262), and network perturbation identified LDHA as the hub. Conclusions. Pyruvate partitioning is cellularly and spatially organized in CRC and defines a myeloid-stromal immunosuppressive niche. Because MSI tumours (the best CRC immunotherapy responders) had the lowest PPI, the PPI-response relationship appears cancer-type specific and requires validation in CRC cohorts.

cancer biology↗

bioRxiv2026

Stressed alga-free coral cells can initiate coral bleaching

Coral bleaching is driving the rapid decline of coral reefs worldwide, yet a fundamental question remains unresolved: does the bleaching cascade begin in the coral host or in its intracellular algal symbionts, Symbiodiniaceae? Resolving where this process starts is critical for understanding how symbiosis collapses and where early intervention might be effective. Here we use transient stress to create a temporal window between the early cellular response and subsequent symbiosis breakdown. Across transient menthol and thermal stress in soft and stony corals, bleaching developed after the stress had ended. In Xenia sp., single-cell transcriptomics revealed a broad host response before detectable symbiont loss, including coordinated redox changes consistent with conditions favouring reactive oxygen species (ROS) accumulation. Cell-resolved imaging showed that alga-free coral cells were the first to become ROS-positive, whereas Symbiodiniaceae showed no detectable ROS signal. This early host response was transmissible. Conditioned medium from stressed cultures induced the emergence of ROS-positive host cells and subsequent symbiosis breakdown in unstressed recipients. Purified ROS-positive alga-free coral cells alone were sufficient to trigger symbiosis breakdown in recipient cultures without direct stress exposure. Suppressing the early emergence of ROS-positive coral cells markedly reduced later bleaching, whereas intervention after stress was substantially less effective. These findings identify a host-initiated, intercellularly propagating route to coral bleaching and reveal an early host response that can determine the subsequent fate of the symbiosis.

cell biology↗

bioRxiv2026

A lifespan-scale single-cell atlas defines an early-childhood immunometabolic transition linked to age-referenced immune states

Childhood represents a critical period with rapid metabolic and immune remodeling, during which perturbations may have lasting consequences for health. We integrated single-cell RNA sequencing (scRNA-seq) profiles of peripheral blood mononuclear cells (PBMCs) from 2,008 healthy individuals aged 0-100 years. Age-resolved modeling highlighted an early-childhood transition centered at age 6, characterized by coordinated remodeling of metabolism-related transcriptional programs, particularly in monocyte and B-lineage cells. Matched serum metabolomics and lipidomics in a pediatric subcohort detected a temporally concordant wave of metabolite and lipid changes around ages 6-8 and extensive metabolite/lipid-transcript associations, prominently involving steroid and phospholipid species. Building on these findings, we developed an immunometabolic transition score and age-specific reference chart, which were evaluated in an independent healthy cohort and applied to quantify deviations in pediatric immune disorders. These findings highlight a coordinated immunometabolic transition in early childhood and provide an age-referenced framework for distinguishing atypical immune-state variation from healthy age-associated variation.

immunology↗

bioRxiv2026

From pose to behavior: SABER integrates identity-resolved multi-animal pose tracking with language-model-based behavioral factor discovery

Quantifying social behavior requires accurate assignment of posture and actions to individual animals, which is often hindered by close contact, occlusion, and identity switches. Meanwhile, current behavioral recognition pipelines still lack stable predictive accuracy. Here we developed SABER, a locally deployable framework that couples multianimal pose estimation with identity preserving tracking, interpretable behavioral factor mining, and multiscale temporal behavior prediction from a single overhead video stream. Across spontaneous two mice social interaction, mating, aggression, and four mice recordings, SABER improved pose estimation accuracy and tracking continuity relative to comparator pipelines. Its behavioral factor-mining procedure identified interpretable kinematic, postural, and social descriptors, and temporal integration improved classification of behavioral categories. SABER offers an intuitive, open-source interface to facilitate use. Applied to social defeat stress mice, SABER detected reduced approach behavior and a multivariate behavioral profile that distinguished depression susceptible from control animals. SABER outputs could also be synchronized with miniscope calcium recordings, enabling joint analysis of behavioral states and neuronal population activity. SABER therefore provides an accessible, identity resolved route from single view social interaction video to behavioral phenotyping and brain behavior analysis.

animal behavior and cognition↗

bioRxiv2026

The Gp78 ubiquitin E3 ligase drives mitophagy by regulating omegasome formation

Autophagy is initiated at endoplasmic reticulum (ER)-associated omegasomes, yet how the ER contributes to mitochondrial cargo degradation during early autophagosome formation remains unclear. Gp78 is an ER-resident RING E3 ubiquitin ligase involved in ER-associated degradation (ERAD) that also promotes Parkin-independent mitophagy. However, the spatial relationship between Gp78 ubiquitin ligase activity, omegasome formation, and mitochondrial degradation has yet to be defined. Combining targeted mitochondria labeling and super-resolution STED microscopy, we show that the Gp78 ubiquitin ligase is localized to and regulates DFCP1-positive omegasome formation, where it drives mitochondrial protein breakdown. Using fluorescent mitophagy reporters and acute ivermectin-induced mitophagy, generation of mitophagy intermediates lacking mitochondrial outer membrane (TOMM20) was found to be dependent on the Gp78 RING finger as well as the BAG6-UBL4A-USP13 ubiquitin-ligase module, and independent of PINK1 and Parkin. Object-based analysis of 3D STED images further showed that both DFCP1- and LC3-positive structures associate primarily with intact mitochondrial fragments which show progressive degradation of mitochondrial content. Gp78 localizes to and promotes, via BAG6, the formation of DFCP1-positive omegasomes and mitochondrial protein degradation in these early autophagic structures. ER-derived omegasome biogenesis is therefore coupled to Gp78-dependent ubiquitin ligase activity and local degradation of mitochondrial cargo during mitophagy.

cell biology↗

bioRxiv2026

PromptBio: An Agentic Platform for End-to-End Computational Biomedical Research

Modern biomedical research increasingly depends on complex computational analyses, yet translating a scientific question into a reliable workflow still requires substantial technical expertise and manual coordination. PromptBio is a multi-agent AI platform available through a web portal at https://promptbio.ai that addresses this challenge. Through natural-language interaction, its agent harness, PromptGenie, translates a research objective into an inspectable plan, executes the required research and analysis, and adapts subsequent steps as evidence and results emerge. PromptBio integrates reasoning with managed execution while preserving human oversight and a traceable record of the research process. It can apply validated methods, construct custom analyses, and incorporate external workflows, enabling researchers to move beyond fixed pipelines while supporting reproducibility. We evaluate the platform through benchmarks of end-to-end bioinformatics analysis and biomedical deep research, validation of representative omics and machine-learning skills, and a hypothesis-driven regulatory-genomics case study. PromptGenie achieved higher analytical accuracy and stronger evidence retrieval and synthesis than the comparison agents, while the evaluated domain skills produced results consistent with established methods. The case study further demonstrates how PromptBio can integrate human-specific genomic and epigenomic features to investigate cortical development. Together, these findings show that PromptBio can coordinate complex biomedical analyses under researcher oversight, offering an accessible approach for accelerating scientific iteration and transforming research questions into transparent, reusable computational workflows.

bioinformatics↗

bioRxiv2026

Rapid spatial cognition in mice, with and without neocortex and hippocampus

Rapid learning, memory, and generalization are often attributed to circuits of the neocortex and hippocampus, but their specific role remains unclear. To examine these cognitive abilities together in individual mice, we observed mice navigating the Manhattan Maze, a reconfigurable 3D labyrinth. Naive wildtype mice improved within two rewards, approached efficient paths within about 20 rewards, retained a 9-turn route overnight, and learned faster in new configurations. Much of the few-shot improvement follows from a rule-based forward bias that emerged even before any reward. In a maze with loops, where that rule is less useful, mice learned within a few rewards to prefer one bottleneck corridor far from the reward location, while choices elsewhere stayed flexible. To begin linking these different components of learning to brain function, we presented the same task to mutant mice that lack the hippocampus and most of the neocortex. They were impaired during initial exploration, where repetitive scanning made them about 3-fold slower to obtain the first few rewards. Past that stage, learning, retention over weeks to months, and generalization to new mazes were largely preserved. Learning this fast is hard to reconcile with trial and error, in which value propagates backwards from the reward. A neuromorphic circuit model instead accounts for the bottleneck choice: it builds a map of the environment without reward, and needs neither cortical nor hippocampal circuit motifs. Therefore, learning is possible without the involvement of neocortex and hippocampus. Structure learned before the first reward, rather than the reward itself, may be what makes few-shot learning possible.

neuroscience↗

bioRxiv2026

A Hymenoptera-restricted gene mediating ant castes co-opts deeply conserved machinery to control organ size

Lineage-specific genes are widespread and have been implicated as phenotypic innovation inducers, but how they acquire complex developmental functions remains poorly understood. Ant queens and workers develop dramatically different organ sizes from identical genomes under juvenile hormone (JH) control, yet the molecular effectors translating JH signalling into caste-specific organ growth remain unknown. Here we identify torch, a Hymenoptera-restricted gene, as the most consistently gyne-biased and JH-responsive gene across 68 ant species. Knockdown of torch in virgin queens of Monomorium pharaonis produces a worker-like, multi-organ growth-restricted phenotype. Mechanistically, torch harbours an E-box-like motif activated by the JH receptor Gce-Tai and acts as a GA-repeat-binding transcription factor that regulates Hippo signalling, the deeply conserved organ-size control pathway in animals. Expressing torch heterologously in mice and a growth-restricted Drosophila background shows that the gene retained its general growth-promoting activity across more than 700 million years of animal evolution in lineages that lack the gene, establishing that its function is mediated through conserved rather than ant-specific machinery. A lineage-specific gene can therefore acquire complex morphogenetic function by co-opting ancient organ-size circuitry, providing a general route by which novel genes can drive phenotypic innovation.

evolutionary biology↗

bioRxiv2026

LAND: Latent Aligned Neural-Behavioral Dynamics via Flow Matching forGeneralizable Movement Decoding

Generalizable movement decoding remains a central challenge for invasive brain--computer interfaces (BCIs), as decoders trained under limited calibration conditions often fail to generalize to unseen movement speeds, limbs, and subjects. Existing decoding methods are typically trained on paired data collected under restricted conditions. How to incorporate behavioral structure from unpaired data for robust out-of-distribution (OOD) decoding therefore remains unresolved. To address this, we propose LAND (Latent Aligned Neural-behavioral Dynamics), a framework that aligns latent neural and behavioral dynamics through flow matching. By learning a neural-to-behavioral transport map and using behavioral-dynamics priors from unpaired data to encourage structured neural manifolds, LAND regularizes representation geometry to promote cross-domain generalization. We evaluate LAND on synthetic neural data, epidural BCI recordings from a tetraplegia participant, and multi-electrode array (MEA) recordings from nonhuman primates (NHPs). Across these settings, LAND improves zero-shot generalization to OOD movement speeds and yields speed-modulated manifolds. With limited target-domain fine-tuning, it further improves transfer across limbs and subjects. These results support flow-based neural--behavioral alignment with unpaired kinematic priors as an approach for learning transferable neural representations and robust movement decoding across behavioral and recording domains.

neuroscience↗

bioRxiv2026

Trehalose exerts cryoprotective effects on piglet testicular tissue

The cryopreservation of testicular tissue is crucial for maintaining male fertility; However, its efficacy is often compromised by oxidative stress and mitochondrial dysfunction. Trehalose, a natural cryoprotectant, demonstrates significant potential, yet its specific mechanisms, particularly in mitochondrial regulation, remain insufficiently characterized. This study aimed to investigate the cryoprotective effects of trehalose on testicular tissue from 18-21-day-old piglets, with a focus on mitochondrial metabolism. Samples were cryopreserved via a slow-freezing protocol in a modified standard solution containing 200 mmol/L trehalose. The protective effect was evaluated by measuring testosterone synthesis, blood testis barrier (BTB) and spermatogenesis. Additionally, protective outcomes were assessed by measuring cell viability, tissue morphology, reactive oxygen species (ROS) levels, apoptosis rates, and testosterone secretion following freeze-thaw cycles. Transcriptomic sequencing and bioinformatics analyses were conducted to elucidate the underlying molecular mechanism. Cryopreservation led to reduced testosterone synthesis and secretion, decreased levels of BTB-binding proteins, and impaired spermatogenesis. Results indicated that 200 mmol/L trehalose significantly improved cell survival, decreased apoptosis and ROS levels, and enhanced testosterone secretion. 200 mmol/L trehalose partially increased the expression of StAR and CYP11A1 genes associated with testosterone synthesis while it protected the tight junction proteins Claudin-11, ZO-1 and the gap junction protein Cx43. Consequently, it exerted a reproductive protective effect by increasing the expression of key spermatogenic regulators DDX25, HMGB2, acrosomal protein DYP19L2, and sperm tail proteins AKAP4 and CFAP44. Transcriptomic profiling demonstrated that trehalose predominantly restored the transcriptional expression of genes involved in the mitochondrial electron transport chain and oxidative phosphorylation pathways, including ND2, COX2, ATP8, ATP6, ND5, ND6 and CYTB. These findings indicate that trehalose primarily protects piglet testicular tissue during cryopreservation by enhancing mitochondrial function, thereby providing a molecular basis for optimizing cryopreservation protocols.

cell biology↗

bioRxiv2026

A genetic switch involving EGR1 and EZH2 links aging and development across human tissues

Aging is often regarded as the last chapter of development. There is growing evidence that aging and development are mechanistically linked. Here we describe a genetic switch involving EZH2 and EGR1 as opposing regulatory nodes that appears to connect aging and development across human tissues. We show that this switch defines two distinct states in fibroblasts: a proliferative (EZH2-high/EGR1-low) state and a non-proliferative and extracellular matrix-expressing (EGR1-high/EZH2-low) state. During replicative aging, cells shift from the proliferative state to the non-proliferative state and targeting either regulatory node--by overexpressing EZH2 or suppressing EGR1--rejuvenates aged fibroblasts. We provide evidence that this switch is involved in the transition from neural progenitor cells to matured neurons during brain development and becomes destabilized during aging, partially reversing the gene expression program established during development. We observed that this same switch becomes blurred and biased towards EGR1-high/EZH2-low state during the aging of muscle stem cells and hematopoietic stem cells. Since genetic switches are widely used in development to establish and reinforce cellular identity, we propose that attenuation of developmental switches may underlie aging across diverse organs and tissues, and partially accounts for the erosion of the epigenetic landscape and the loss of cellular identity.

systems biology↗

bioRxiv2026

FXR-mediated recruitment of PPP1CB suppresses SMAD2/3 phosphorylation to mitigate pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal interstitial lung disease with limited treatment options and poorly understood molecular underpinnings. Dysregulated TGF-{beta}/SMAD signaling is a key driver of fibrotic remodeling, promoting persistent myofibroblast activation and excessive extracellular matrix deposition. Here we identify the Farnesoid X receptor (FXR), a bile acid activated nuclear receptor, as a previously unrecognized suppressor of pulmonary fibrosis. FXR expression is significantly reduced in lung tissues from patients with PF and in myofibroblasts derived from BLM-induced mouse models, correlating inversely with fibrosis severity. Genetic ablation of FXR exacerbates BLM-induced pulmonary fibrosis by promoting fibroblast hyperactivation and dysregulation of the TGF-{beta}/SMAD signaling pathway. Mechanistically, we identify PPP1CB as a previously unrecognized FXR-interacting protein in primary myofibroblasts derived from IPF patients. We further show that FXR both increases chromatin accessibility at the PPP1CB locus and assembles a functional complex with PPP1CB, which in turn promotes SMAD2/3 dephosphorylation and suppresses their nuclear translocation. Notably, the clinical-stage FXR agonist TERN101 exhibits potent therapeutic efficacy in a BLM-induced mouse pulmonary fibrosis model. These findings establish FXR as a critical antifibrotic regulator in lung tissue and suggest that pharmacological activation of FXR may offer a promising therapeutic strategy for IPF.

cell biology↗

bioRxiv2026

Drought duration does not impact soil microbiome resilience

Increasing global droughts exert large but poorly understood effects on the microbial communities and ecology of soil. Microbial communities generally show resilience and return to pre-drought conditions when short-term droughted soils are rewet; soils exposed to long-term drought, however, often show a lag upon rewetting, after which microbial communities may or may not return to their pre-stressed conditions. Though short-term droughts have been widely studied, long-term drought manipulation experiments remain rare, especially those that compare microbial response to short-term and long-term drought in tandem. We conducted a 1000-day drought simulation in controlled laboratory conditions with soil cores collected from a tidal freshwater ecosystem in Washington state, USA, and subsequently exposed them to rewetting for two weeks. We also included short-term (30-day and 90-day) drought and rewet treatments to directly compare microbial community and organic matter responses across drought durations. We found distinct microbial taxa belonging to Firmicutes and Actinobacteria enriched after the 1000-day drought, but not after the short-term droughts. While we hypothesized that the microbial community would recover from a short-term drought after rewetting to resemble pre-drought conditions, our results revealed community dissimilarities between rewet and pre-drought conditions across all drought durations. These findings suggest unique microbial life history strategies within certain microbial phyla that make them successful colonizers during an extended drought period, and the influence of environmental and physiological context on microbial responses to rewetting. ImportanceDroughts are increasing in frequency and intensity globally with severe implications for ecosystem services and soil functions. It is important to understand how long-term drought impacts soil microbial communities and organic matter chemistry to better predict future ecosystem responses to sustained moisture deficit conditions. We subjected soils to short-term (30 and 90 days) and long-term (1000 days) drought treatments and subsequently rewetted them to understand microbiome recovery to pre-drought conditions. Our results showed that prolonged drought drastically changes the microbial community and soil organic matter profile compared to short-term drought. While we expected the soil microbiome to recover upon rewetting after short-term drought, our results showed an altered microbiome composition, compared to pre-drought conditions, for both short-and long-term drought, suggesting microbial responses to soil rewetting was independent of drought duration imposed. These results provide important insights into soil biological and chemical functions that remain sensitive to change under fluctuating soil moisture conditions and future drought scenarios.

ecology↗

bioRxiv2026

A novel AR molecular glue degrader via bivalent engagement of the N-terminal domain and RNF213-UBE2J2 complex

Targeted protein degradation enables the elimination of disease-relevant proteins through induced proximity between E3 ligases and substrates. Current androgen receptor (AR) therapies act via the ligand-binding domain (LBD) but fail against constitutively active LBD mutants or splice variants such as AR-V7. Here, we identify GZL626, a small-molecule degrader that induces proteasome-dependent degradation of both full-length AR and AR-V7. GZL626 suppresses AR transcriptional activity, inhibits proliferation of AR-positive prostate cancer cells, and blocks SARS-CoV-2 infection. Mechanistically, GZL626 binds the intrinsically disordered AR N-terminal domain (NTD) and promotes formation of a noncanonical RNF213-UBE2J2-AR ternary complex that drives AR ubiquitination and degradation. The AR DNA-binding domain (DBD) acts as a structural hub within this assembly. Distinct from PROTACs or classical molecular glues, GZL626 engages both AR and RNF213 bivalently, remodeling protein interactions to stabilize a functional E3-E2-substrate complex. These findings establish GZL626 as a first-in-class NTD degrader with potential to overcome resistance in advanced prostate cancer.

biophysics↗

bioRxiv2026

Multimodal computational framework identifies B cell convergence in autoimmunity and ageing

Identification of the origin of pathogenic immune cells is crucial for therapeutic interventions and diagnosis but pseudotime methods struggle to trace immune cells accurately. Current trajectory inference methods for B cell development and response in health and disease either ignore or underutilize antigen receptor sequence information, limiting their ability to resolve developmental pathways, particularly for pathogenic populations. Widely used methods such as Monocle 3 reconstruct developmental paths from transcriptomic similarity alone, discarding the features from immune receptors. Dandelion has combined the immune receptor features with transcriptomics but it struggles to simulate the trajectory path of B cells. Here we present ClonoTrace, a computational framework that integrates BCR sequence features with transcriptomic trajectory inference through gated fusion of multimodal embeddings. In fetal B cell development and germinal centre development, ClonoTrace achieves higher trajectory inference accuracy than Monocle 3 and Dandelion. Applied to systemic lupus erythematosus, ClonoTrace identifies memory B cell extrafollicular maturation pathway in addition to naive B cell, accompanied by induction of ZEB2 with a concomitant decline of BACH2 along the trajectory, as the alternative origin of pathogenic double negative 2 B cells (DN2) in systemic lupus erythematosus (SLE) patients. In healthy ageing, ClonoTrace identified three pathways from naive, IgM+ memory B cells and switched-memory B cells mature through a DN2-associated transcriptional state that precedes age-associated B cells. ClonoTraces fate probability algorithm indicated that IgM+memory B cell to ABC transition emerged as the leading candidate age-associated transition, that is a process distinct from SLE DN2 maturation. ClonoTrace provides a generalizable framework for receptor-informed trajectory inference, revealing the developmental pathways of pathogenic B cell populations that are untraceable to single modality approaches in autoimmunity and aging.

bioinformatics↗

bioRxiv2026

Near-Infrared II Scintillator for High-Resolution X-ray Imaging

Light scattering in scintillators is a pervasive problem and a key factor limiting X-ray imaging resolution. Here, we shift scintillator radioluminescence from the traditional visible range into the short-wave infrared (SWIR) or near-infrared II (NIR-II, 1000-3000 nm) window to mitigate light scattering and thereby enhance light penetration and X-ray imaging resolution. We present an NIR-II MgGa2O4:Ni2+ scintillator with peak emission at 1340 nm, achieving a threefold improvement in X-ray imaging resolution compared with visible scintillators owing to reduced light scattering. This heavy-metal-free NIR-II scintillator exhibits intense radioluminescence comparable to that of conventional visible-emitting CsI:Tl, achieving a detection limit of 56 nanograys per second, [~]100-fold lower than typical doses used in medical imaging. We show that this NIR-II scintillator enables high-resolution X-ray radiography of electronic circuit boards and biological tissues.

biophysics↗

bioRxiv2026

Label-free Pathogen Identification with Microscopy Imaging and Deep Learning

Rapid and accurate pathogen identification is crucial for the clinical management of infectious diseases, particularly sepsis and severe respiratory infections, yet standard clinical workflows remain slow and resource-intensive. Here, we developed an automated, high-throughput imaging platform built on standard, clinically accessible bright-field microscopy, and generated a large dataset comprising 24.9 million label-free bacterial cells across six focal pathogens. Leveraging this resource, we trained a neural network (ESKAPe-ResNet) to identify ESKAPe species at the single-bacterium level. The model achieved >92% accuracy in species-level classification and >82% accuracy in quantifying ESKAPe abundance in mock mixtures, with high specificity against non-ESKAPe bacteria. In clinical validation using sputum, bronchoalveolar lavage fluid and blood samples from patients with respiratory infections and sepsis, the approach correctly identified the dominant ESKAPe pathogen in >78% of samples after minimum broth culture enrichment. The imaging-to-identification pipeline was completed in under 10 minutes, and coupled with brief cultivation, the median time to accurate identification was reduced to 5-6 hours, compared with days for conventional blood culture-based workflows. This work establishes the proof-of-principle for label-free, hardware-minimal rapid pathogen identification, providing a clinically deployable workflow to expedite diagnosis and reduce mortality in severe bacterial infections.

microbiology↗