Biology subjects

Ito, M.

Publications and source records attributed to Ito, M..

At least 19 recordsLinked to original sources

bioRxiv2026

Reverse genetics system for emerging tick-borne orthonairovirus highlights dispensable N-terminal region of Gn associated with replication in tick vectors

A growing number of emerging and re-emerging tick-borne orthonairoviruses belonging to the Sulina and Tamdy genogroups have recently been identified in association with febrile diseases in East Asia. Yezo virus (YEZV) is one such viruses and is genetically distinct from well-characterized Crimean-Congo hemorrhagic fever virus. Orthonairovirus glycoproteins (Gn and Gc) expressed from a single glycoprotein precursor (GPC) gene mediate interaction with host cells, yet the organization and functions of the YEZV glycoproteins remain largely undefined. To characterize YEZV glycoproteins in the context of infection, we developed a reverse genetics system that enables recovery of recombinant YEZV entirely from cloned cDNAs. The recombinant wild-type virus exhibited growth properties comparable to those of the parental isolate in vitro and maintained pathogenicity in vivo. Proteomic analysis of purified virus particles produced in mammalian cells showed that peptide coverage of GPC-derived products began at residue 69. We therefore used the reverse genetics system to examine the functional importance of the GPC subregion upstream of residue 69. A mutant with a deletion of GPC residues 28 to 68 (rYEZV GPC{Delta}28-68), which retained the predicted signal peptide, was successfully recovered, suggesting that this subregion is dispensable for producing infectious virus. rYEZV GPC{Delta}28-68 propagated at levels comparable to those of the wild-type virus in mammalian cells and exhibited similar pathogenicity in a mouse model. Interestingly, the mutant reached lower viral titers in tick-derived ISE6 cells and in ticks in vivo, suggesting that this N-terminal subregion may contribute to virus replication in ticks. Together, these findings demonstrate the utility of the newly established reverse genetics system for dissecting the functions of the YEZV glycoproteins in mammalian and tick systems.

microbiology↗

bioRxiv2026

REN-former prioritizes candidate regulators of kidney disease-state transitions through single-cell foundation modeling and human genetics

Background Acute kidney injury (AKI)-to-chronic kidney disease (CKD) transition is associated with dynamic changes in tubular cell state. However, conventional single-cell transcriptomic analyses primarily identify genes differentially expressed between disease states and do not directly evaluate genes associated with directional transitions between cellular states. Methods We developed REN-former by fine-tuning Geneformer using the GSE183276 single-cell RNA-sequencing dataset from 45 participants representing Normal Reference, AKI, and CKD states. In silico gene deletion and overexpression analyses were used to estimate directional transcriptomic shifts, with a focus on proximal tubular cells. Selected genes were evaluated using summary-data-based Mendelian randomization (SMR), colocalization and expression analysis in additional KPMP participants not included in GSE183276. Results REN-former achieved recall values of 0.99, 0.80, and 0.79 for Normal Reference, AKI, and CKD, respectively. In silico perturbation analyses identified distinct gene programs associated with transitions from Normal Reference to AKI, from Normal Reference to CKD, from AKI to CKD, and from CKD to Normal Reference. Conventional analysis showed metabolic suppression and increased inflammatory and stress-response activation. SMR identified IFITM3, CALR, TTR, CALM1, MUC13, and RPL13, and colocalization supported IFITM3, CALR, TTR, and CALM1. In additional KPMP data, IFITM3 was higher, whereas TTR and CALM1 were lower, in CKD proximal tubules; CALR did not differ significantly. The observed expression changes were concordant with the REN-former-predicted directions for TTR and CALM1 but discordant for IFITM3. Conclusion REN-former provides a framework for prioritizing candidate regulators of kidney disease-associated cell states by integrating predicted perturbation effects with human genetic and transcriptomic evidence.

bioinformatics↗

bioRxiv2026

Prepatterned Tissue Stiffness Gradient Controls Organ shape and size

Organ morphogenesis is orchestrated by precise coupling of mechanical and biochemical signals, yet how these two signals are integrated within tissue niche to regulate organ shape and size remains poorly understood. Here, using sweat gland as a tractable model of exocrine mini-organs, we demonstrate that dermal fibroblasts play a key role to translate prepatterned tissue stiffness gradients into spatial Wnt5a gradients via Piezo1-mediated mechanosensing. Following epidermal placode formation, progenitor cells collectively invaginate into the dermis, where spatiotemporally graded Wnt5a directs their sequential transition from ductal to glandular fate in softer distal dermis, specifying distinct exocrine compartments. Perturbation of dermal stiffness gradients and mechanosensing in vivo and ex vivo alters Wnt5a expression levels and gradient patterns and thereby changes the ductal length and glandular size, revealing a direct link between tissue mechanics and organ dimensions. To recapitulate this process in vitro, we engineer a microfluidic platform capable of generating robust and linear morphogen gradients in 3D, allowing duct-to-gland fate transitions to occur at a high resolution. Together, our finding identifies prepatterned stiffness gradients as a master upstream regulator of coupled mechanical-biochemical signaling, define a mechanistic framework for two-step sequential glandular morphogenesis, and offer new strategies for engineering glandular epithelia in regenerative medicine.

developmental biology↗

bioRxiv2026

Evaluation of Locomotor Activity in Group-Housed Hamsters: Effects of Cage Size and Running Wheel Availability

While hamsters are commonly housed in groups within pet shops in Japan, small cages are often thought to restrict their physical activity, leading to arguments that larger cages should be provided. Although previous research has investigated how cage size and running wheel availability influence activity levels in individually housed hamsters, no studies to date have examined these specific effects in a social housing context. Therefore, this study investigated how cage size and the presence of a running wheel affect the activity levels of individual hamsters using the group-housing conditions with five hamsters. Video recordings were captured for 24 hours across four distinct cage environments using a camera installed directly above each cage. From these recordings, the distances traveled on both the cage floor and the running wheel were calculated for each hamster and statistically analyzed. The results revealed that overall activity levels were significantly higher in cages equipped with a running wheel than in those without. Although cage size did not yield a statistically significant difference, a marginal trend toward higher activity in larger cages was observed.

animal behavior and cognition↗

bioRxiv2026

A rapidly deployable CRISPR-Cas3 diagnostic platform for emerging RNA viruses

Rapidly converting viral genome information into deployable molecular tests remains a major challenge in outbreak preparedness. We developed CONAN-SWIFT (Simple Workflow for Isothermal Field Testing), a sequence-to-test platform that integrates computational assay design, reverse-transcription loop-mediated isothermal amplification, CRISPR-Cas3 detection, reagent lyophilization and lateral-flow readout. Sequence-guided assays for Andes virus and Bundibugyo virus were established within approximately three weeks and extended to four additional filoviruses. A web-based designer supported crRNA selection, and systematic RT-LAMP primer optimization improved amplification performance. Recombinant Escherichia coli-expressed Cascade enabled standardized preparation of lyophilized Cas3-detection reagents, which were combined with a battery-operated isothermal device. The portable system detected as few as 10 input RNA copies per reaction within approximately 40 min. It also detected viral RNA and biologically contained, replication-incompetent Ebola virus in spiked human blood and concentrated wastewater. These findings establish the analytical feasibility of a rapidly adaptable CRISPR-Cas3 engineering framework for decentralized detection of emerging RNA viruses.

bioengineering↗

bioRxiv2026

Multi-layered control of chromosomal assembly of the meiotic DNA break machinery

Meiotic DNA double-strand breaks (DSBs) form in chromatin loops that are tethered to chromosome axes. How proteins required for DSB formation in vivo (DSB proteins) assemble on meiotic chromosomes to control non-random DSB distribution remains unsolved. Here we mapped the spatial distributions of all ten DSB proteins that form three complexes (Spo11-Ski8-Rec102-Rec104 [Spo11-core], Rec114-Mei4-Mer2 [RMM], and Mre11-Rad50-Xrs2 [MRX]) in budding yeast and elucidated their functional dependencies with axial proteins Red1 and Hop1. At local levels, our analysis suggests that their axis-associated assembly involves two distinct pathways, Hop1-RMM-MRX and Red1-Spo11 core, whereas Spo11-core alone can bind DSB sites. Red1 also binds DSB sites in a DSB-dependent manner, supporting its critical role in DSB repair. At large scale, all ten DSB proteins and both axial proteins are enriched at DSB-hot domains 20-40 kb wide that correspond to sizes of the axis-loop units. Further analysis at short and long distances suggests that their local assembly, short-range/intra-loop distributions, and long-range/inter-loop distributions are distinctly controlled. Similarly in mouse spermatocytes, MEl4 and a Hop1 homolog HORMAD1 are enriched at DSB-hot domains 1-3 Mb wide, and their long-range distributions are distinctly controlled from their local assembly. Our results elucidate multi-layered mechanisms of chromosomal assembly of the DSB machinery in shaping the DSB landscape in yeast and mouse.

Molecular Biology↗

bioRxiv2026

Analysis of publicly available genomic data

Ethambutol (EMB) is a vital drug for treating Mycobacterium avium-intracellulare (MAI) infections; however, the genomic mutations underlying EMB resistance in MAI remain unclear. Herein, we evaluated eight sets of MAI clinical isolates, each containing at least two serial isolates collected from the same patient who received EMB in Japan. In four sets, the isolates independently increased EMB MIC by 4-fold, coinciding with mutations in the upstream region of embA or those corresponding to Mycobacterium tuberculosis (Mtb) embB Met306Val and Gln497Arg. Based on the increased EMB MIC values, we defined normal and elevated EMB MICs as [≤]8 {micro}g/mL and [≥]16 {micro}g/mL, respectively. In the other four sets, all of the isolates had elevated EMB MICs. In silico promoter prediction and expression analysis indicated that the upstream region of embA corresponds to the embA-embB promoter region, and mutations in this region increased the transcription of embA and embB, increasing EMB MICs. Furthermore, the analysis of 60 epidemiologically unrelated strains revealed that isolates with mutations in the embA-embB promoter and at embB codons 306/497 exhibited significantly higher EMB MICs compared with those without mutations. Publicly available genomic data demonstrate the worldwide occurrence of these mutations in clinical isolates. These results establish an association between elevated EMB MICs and mutations at embB codons 306/497 and the embA-embB promoter and are expected to predict EMB resistance.

microbiology↗

bioRxiv2026

Single-cell spatial multiomics identifies POSTN+ CAFs mediating chemoradiotherapy resistance in rectal cancer

Neoadjuvant chemoradiotherapy (CRT) is standard for locally advanced rectal cancer (LARC), yet many patients retain residual disease. To resolve CRT-associated remodeling of the tumor microenvironment, we generated a multimodal spatial atlas from serial sections of paired pretreatment and post-treatment specimens from 24 patients using Xenium single-cell spatial transcriptomics and PhenoCycler multiplex proteomics, profiling 2.8 million cells; matched Visium HD datasets were generated on adjacent serial sections. Resistance was most strongly associated with fibroblast and myeloid programs adjacent to residual tumor. We identify a periostin (POSTN)-expressing CAF subset selectively enriched around residual tumor cells in non-responders, displaying a myofibroblastic phenotype and activating extracellular matrix remodeling, noncanonical WNT signaling, and immunosuppressive pathways. Tumor cells neighboring POSTN+ CAFs show consistent epithelial-mesenchymal transition signatures. Together, this atlas enables interrogation of CRT-induced spatial remodeling and nominates POSTN+ CAFs as key mediators and targets of CRT resistance, with direct relevance to CRT-based combination strategies.

cancer biology↗

bioRxiv2026

Identification of scavenger receptor BI as a scavenger of free heme that is essential for protection against hemolysis

Severe hemolysis is a life-threatening condition with limited therapeutic options. Although haptoglobin and hemopexin sequester hemoglobin and heme, these protective systems are rapidly saturated during acute hemolysis, leading to the accumulation of cytotoxic free heme. In this study, we identify scavenger receptor BI (SR-BI) as a critical mediator of free heme clearance. SR-BI binds heme and facilitates its hepatic uptake under pathological conditions. Mice lacking hepatic SR-BI exhibit impaired heme clearance and increased susceptibility to heme- and hemolysis-induced lethality. Pharmacological upregulation of hepatic SR-BI via imatinib or adenoviral delivery confers protection against heme toxicity. Using a humanized model of sickle cell disease (SCD), we further demonstrate that sickle hepatopathy significantly reduces hepatic SR-BI expression compared to non-SCD littermates, potentially increasing vulnerability to heme-induced injury. Notably, adenoviral-mediated SR-BI upregulation rescues SCD mice from heme toxicity. These findings reveal a previously unrecognized mechanism of heme detoxification via hepatic SR-BI and identify a promising therapeutic target for hemolytic disorders. One-Sentence SummaryIdentification of scavenger receptor BI as a targetable scavenger of heme in hemolysis

physiology↗

bioRxiv2026

Efficient generation of hematopoietic progenitor cells from human pluripotent stem cells by robotic automation

Differentiation of human pluripotent stem cells (hPSCs) into therapeutic cell types requires stringent control of developmental signals, but conventional manual operations introduce substantial intra- and inter-operator variability. While automation can reduce such variability, fixed systems often lack compatibility with the dynamic, stage-specific, and format-diverse manipulations required for hPSC culture and differentiation. Here, we establish a flexible robotic platform to standardize a stroma- and xeno-free embryoid body (EB)-based hematopoietic progenitor cell (HPC) differentiation process and integrate machine learning (ML) to optimize key process parameters including the concentrations of multiple signaling molecules. Despite intrinsic biological stochasticity and technical complexity associated with EB formation, the robotic platform substantially reduced experimental variation. ML-guided, unbiased exploration not only identified combinations of signaling inputs that markedly improved the efficiency and reproducibility of HPC differentiation and subsequent natural killer cell generation, but also illuminated key signaling logic underlying early human hematopoietic development. Together, these results demonstrate that data-driven robotic automation can uncover optimal culture conditions that are difficult to identify through conventional human-driven experimentation.

developmental biology↗

bioRxiv2026

Age-dependent accumulation of RAD51 on non-damaged chromosomes prevents chromosome segregation in mammalian oocytes

RAD51 is targeted to single-stranded (ss)DNA for homologous recombination and DNA replication fork homeostasis. However, the physiological consequences of RAD51 binding to intact double-stranded (ds)DNA, which is tightly limited in vivo, remain elusive. Here we revealed an intrinsic property of RAD51 to bind chromosome axes where cohesin and condensin bind, which is actively suppressed by FIGNL1 AAA+ ATPase. In Fignl1-deficient mouse oocytes, an age-dependent RAD51 accumulation with little DNA damage leads to improper chromosomal localization of condensin II and topoisomerase II, failure in chromosome condensation with massive chromosome entanglement, and meiosis I arrest. We propose that promiscuous RAD51 binding to non-damaged chromosomes, which is prevented by a RAD51 remodeler, is a unique type of chromosomal pathology associated with genome instability.

cell biology↗

bioRxiv2026

Spatial distribution of ticks and tick-borne pathogens in central Hokkaido, Japan and associated ecological factors revealed by intensive short-term survey in 2024

Tick-borne pathogens are transmitted by tick bites to cause infectious diseases in humans and domestic animals. To anticipate the occurrence of tick-borne diseases, it is required to understand high resolution distribution of infection risk and associated ecological factors. The aim of the present study is to reveal the spatial distribution of ticks and ticks infected with pathogens in central Hokkaido, Japan. Adult and nymphal ticks were collected with a constant effort at 171 sites from 13 May to 26 June 2024, followed by screening tick-borne pathogens. The potential suitable habitats of seven tick species and the endemic tick-borne pathogens in the study area (i.e. tick-borne encephalitis virus, Yezo virus, Beiji nairovirus, Lyme disease group borreliae, and relapsing fever group borreliae) were predicted using ecological niche modeling. Ixodes persulcatus and Ixodes ovatus were identified as the primary ticks to determine the distributions of all the pathogens. Besides, the predicted suitable habitats were specific to each pathogen/tick species. Among the environmental variables considered for modeling, snow depth appeared to significantly contribute to the distribution differences between ticks and pathogens. The findings of this study expand our understanding of the spatial risk distribution of tick-borne pathogen infections and its ecological context.

ecology↗

bioRxiv2026

The REFLEX system enables in vivo identification of perivascular angiogenic macrophages in the heart

Direct identification of physically interacting cells in vivo remains challenging because conventional interactome analyses infer signaling partners from transcriptomes and cannot reveal which cells are in direct contact. In pressure-overload induced cardiac remodeling, VEGF-A plays a central role in the maintenance of vascular integrity and cardiac function. However, the cell type which produces VEGF-A and how the VEGF-A peptide is delivered to vascular endothelial cells remains unclear. Here, we developed a genetically encoded platform that combines REFLEX mice with HUNTERuni-seq, enabling unbiased detection and transcriptional profiling of the cells that physically interact with vascular endothelial cells. The REFLEX and HUNTERuni-seq approach identified subpopulations of Vegfa positive macrophages which we named perivascular angiogenic macrophages (PVAMs). Although the amount of VEGF-A in PVAMs is small, loss of VEGF-A in PVAMs impaired angiogenesis and systolic function during pressure overload. We additionally show that direct contact between PVAMs and endothelial cells is critical for the delivery of VEGF-A to endothelial cells. Conventional interactome analysis predicted that cardiomyocytes as dominant sources of VEGF-A in the heart. However, cardiomyocyte Vegfa deletion had no effect on capillary density nor systolic function in a model of heart failure. These results suggest that VEGF-A signaling does not rely on free diffusion through the interstitium and that cellular proximity and physical contact between PVAMs and endothelial cells are the key determinants of effective signal delivery. Together, these findings establish REFLEX and HUNTERuni-seq as a versatile platform for uncovering biologically critical cell-to-cell interactions and provide new insight into intercellular communication in pathological tissue contexts.

cell biology↗

bioRxiv2026

A lethal mouse model of Oz virus infection reveals hepatic involvement and enables evaluation of antiviral and vaccine efficacy

Oz virus (OZV), a member of the genus Thogotovirus in the family Orthomyxoviridae, is an emerging tick-borne virus reported in Japan. A fatal human case and seroepidemiological evidence of widespread exposure among wild animals and humans suggest its potential public health significance. However, no animal models suitable for pathogenic studies or evaluation of countermeasures are available for OZV. Here, we have established a lethal mouse model of OZV infection using cell-adapted virus and mice lacking type I interferon signaling (B6 Ifnar1 KO mice). OZV infection resulted in 100% mortality and was characterized by robust viral replication in the liver and spleen, severe hepatitis, and acute liver injury. Using this model, we also demonstrated that oral administration of T-705, an antiviral drug widely used against RNA viruses, as well as immunization with an inactivated whole virus particle vaccine, protected B6 Ifnar1 KO mice from lethal OZV infection by mitigating the acute hepatitis. The present study provides critical insights into OZV pathogenesis and establishes a practical in vivo platform for the development of countermeasures against OZV infection. Significance statementEmerging tick-borne viruses pose a growing public health concern, yet progress in understanding their pathogenesis and in developing countermeasures is often limited by the lack of relevant animal models. Oz virus (OZV), a recently identified thogotovirus associated with a fatal human case, exemplifies this challenge. Here, we establish a lethal mouse model of OZV infection that reveals pronounced hepatic involvement as a central pathological feature. Using this model, we demonstrate effective protection by both an antiviral drug and an inactivated vaccine against lethal OZV challenge. This study provides a practical in vivo platform for investigating OZV pathogenesis and for accelerating the development of medical countermeasures against this emerging tick-borne virus.

microbiology↗

bioRxiv2026

Large-scale brain network differences during deliberate and spontaneous mind-wandering in a sustained attention task: An electroencephalography source-level connectivity study

Mind-wandering (MW), the shift of attention away from an ongoing task, can be classified as either deliberate or spontaneous, depending on whether these internal thoughts arise intentionally or unintentionally. Previous functional magnetic resonance imaging-based resting state studies have shown that spontaneous MW traits are positively correlated with coupling within the default mode network (DMN), whereas deliberate MW traits are positively correlated with coupling both within the DMN and between the DMN and control-or attention-related networks. However, state-based functional connectivity during each MW episode remains unknown. Addressing this knowledge gap, we investigated how deliberate and spontaneous MW differentially involve state-based large-scale network interactions during a sustained attention task using electroencephalography (EEG), which captures rapid network dynamics. Thirty-one participants performed the gradual-onset continuous performance task using 64-channel EEG. Mental states were classified via experience-sampling probes as on-task, deliberate, or spontaneous MW. EEG data from 1-s pre-probe epochs were analyzed using source estimation and the weighted phase lag index to assess functional connectivity within and across the DMN, control network (CN), dorsal attention network (DAN), and salience network (SN) in the delta, theta, alpha, and beta frequency bands. Relative to both spontaneous MW and the on-task state, deliberate MW was characterized by significantly stronger alpha band functional connectivity. A prominent connectivity cluster was centered on the right frontal operculum-insula of the SN, linking regions across the SN, CN, and DMN. In addition, deliberate MW exhibited enhanced SN-DAN-DMN coupling compared with the on-task state. By contrast, spontaneous MW did not show significant connectivity differences relative to the on-task state in any frequency band. Taken together, these results suggest that alpha band network interactions centered on the SN may contribute to the intentional regulation of internally directed thought during deliberate MW.

neuroscience↗

bioRxiv2025

A SABATH family enzyme regulates development via the gibberellin-related pathway in the liverwort Marchantia polymorpha

The SABATH family enzymes are a group of plant-specific methyltransferases that catalyze the methylation of many small molecules, including several plant hormones. While this family emerged anciently before the evolution of land plants from streptophyte algae, little is known about their biological function in plant lineages other than angiosperms. Here, we identified 12 SABATH family genes from the liverwort Marchantia polymorpha and found that MpSABATH2 is essential for its development. Mpsabath2 mutants were severely inhibited in thallus growth and gemma cup formation, while spontaneously forming sexual branches under non-inductive conditions. These phenotypes resembled the developmental responses to far-red light, which was also supported by transcriptome analysis. Further genetic analysis connected this phenomenon with gibberellin (GA)-related metabolism. Blocking GA biosynthesis partially rescued Mpsabath2 phenotypes, which were restored by treatment with the GA precursor, ent-kaurenoic acid. Given that bryophyte and angiosperm SABATH proteins fall into distinct phylogenetic clades, our findings suggest that SABATH family enzymes independently acquired roles in developmental regulation through convergent or parallel evolution in land plants.

plant biology↗

bioRxiv2025

From perception to valence: a pair of interneurons that assign positive valence to sweet sensation in Drosophila

Assigning valence--appeal or aversion--to gustatory stimuli and relaying it to higher-order brain regions to guide flexible behaviors is crucial to survival. Yet the neural circuit that transforms gustatory input into motivationally relevant signals remains poorly defined in any model system. In Drosophila melanogaster, substantial progress has been made in mapping the sensorimotor pathway for feeding and the architecture of the dopaminergic reinforcement system. However, where and how valence is first assigned to a taste has long been a mystery. Here, we identified a pair of subesophageal zone interneurons in Drosophila, termed fox, that impart positive valence to sweet taste and convey this signal to the mushroom body, the flys associative learning center. We show that fox neuron activity is necessary and sufficient to drive appetitive behaviors and can override a tastants intrinsic valence without impairing taste quality discrimination. Furthermore, fox neurons transmit the positive valence to specific dopaminergic neurons that mediate appetitive memory formation. Our findings reveal a circuit mechanism that transforms sweet sensation into a reinforcing signal to support learned sugar responses. The fox neurons exhibit a convergent-divergent "hourglass" circuit motif, acting as a bottleneck for valence assignment and distributing motivational signals to higher-order centers. This architecture confers both robustness and flexibility in reward processing--an organizational principle that may generalize across species.

neuroscience↗

bioRxiv2025

Elevated plasma cholesterol causally improves sepsis outcome by promoting hepatic metabolic reprogramming

RationaleSepsis is a life-threatening condition with high mortality and limited therapeutic options. While hypocholesterolemia is associated with poor outcomes, the causal role of cholesterol and its underlying mechanisms remain unclear. ObjectivesTo determine whether elevated plasma cholesterol improves sepsis survival and to elucidate the mechanistic basis of this effect. MethodsWe analyzed septic patients from the MIMIC-IV database. Total cholesterol levels were compared between 28-day survivors and non-survivors using Wilcoxon rank-sum tests. Cox proportional hazards regression assessed the association between cholesterol levels and mortality, adjusting for age, sex, race, SOFA score, and Charlson comorbidity index. To establish causality, C57BL/6J mice were randomized to receive either a high-cholesterol diet (HCD) or regular diet (RD) for three days prior to cecal ligation and puncture (CLP) to induce sepsis. Measurements and Main ResultsSurvivors had significantly higher cholesterol levels than non-survivors (median, 135 vs. 126 mg/dL, p < 0.001). High cholesterol ([≥] 133 mg/dL) was independently associated with reduced 28-day mortality (adjusted HR = 0.80, 95% CI: 0.67-0.95, p = 0.012). In mice, HCD elevated plasma cholesterol and significantly improved survival (52.5% to 90%), independent of immune modulation.Hepatic transcriptomics revealed metabolic reprogramming, including upregulation of oxidative phosphorylation and glutathione-mediated antioxidant pathways, and suppression of endoplasmic reticulum proteostasis. Notably, inhibition of mitochondrial respiration abolished the survival benefit. ConclusionsElevated plasma cholesterol causally improves sepsis outcomes by promoting hepatic metabolic reprogramming. These findings offer mechanistic insight into clinical observations and suggest hepatic bioenergetics as a novel therapeutic target in sepsis.

physiology↗