Biology subjects

Yin, X.

Publications and source records attributed to Yin, X..

At least 19 recordsLinked to original sources

bioRxiv2026

CAR-macrophage immunotherapy targeting fibroblast-like synoviocytes ameliorates inflammatory and degenerative arthritis

Rheumatoid arthritis (RA) is an inflammatory arthritis, whereas osteoarthritis (OA) is a degenerative joint disease. Despite their distinct etiologies, both are characterized by the pathogenic activation of fibroblast-like synoviocytes (FLS), which contribute to synovial hyperplasia, persistent inflammation, and cartilage destruction. Accordingly, pathogenic FLS represent an attractive therapeutic target for both RA and OA. Macrophages exhibit remarkable phenotypic plasticity, and interleukin-4 (IL-4) promotes an anti-inflammatory, tissue-reparative M2-like phenotype. Fibroblast activation protein (FAP) and nucleolin (NCL) are highly expressed on the surface of pathological FLS while exhibiting limited accessibility in normal tissues. Here, we developed two IL-4-armored, FLS-targeted chimeric antigen receptor macrophage (CAR-M) platforms directed against FAP or NCL, termed CAR-M2-FAP and CAR-M2-NCL, respectively. In vitro, both platforms enhanced FLS phagocytosis and promoted an M2-like transcriptional profile. Transcriptomic analysis further revealed enrichment of phagocytic, anti-inflammatory, and tissue-reparative programs. In a mouse model of RA, both CAR-M2 platforms attenuated arthritis severity, synovial hyperplasia, cartilage loss, and bone erosion. Similarly, in a mouse model of OA, they reduced synovitis, cartilage degeneration, and osteophyte formation. No overt acute toxicity was observed in either RA or OA mice following treatment with CAR-M2-FAP or CAR-M2-NCL. Collectively, the concordant therapeutic effects of CAR-M2-FAP and CAR-M2-NCL across RA and OA models provide preclinical proof of concept for FLS-targeted CAR-M2 as a potential therapeutic strategy for inflammatory and degenerative arthritis.

bioengineering↗

bioRxiv2026

Astatine-211 injury induces an immunostimulatory melanoma-cell state and promotes systemic antitumor immunity

Targeted alpha therapy is generally evaluated by absorbed dose and direct tumor-cell killing, but whether alpha-particle-injured tumor cells can induce systemic antitumor immunity remains poorly defined. We investigated the immunogenic consequences of astatine-211 (211At) injury in a prophylactic melanoma vaccination-challenge model. B16F10 cells were exposed ex vivo to 211At, etoposide, or ultraviolet irradiation under conditions that reduced viable-cell counts at 24 hours to approximately 40% of untreated controls. Injured cells were administered subcutaneously to syngeneic C57BL/6J mice before intravenous challenge with viable B16F10 cells. Vaccination with cells exposed to 1 MBq/mL 211At suppressed pulmonary metastasis, reduced regulatory T-cell proportions, and elicited tumor-reactive interferon-{gamma} responses and antibodies. Transcriptomic profiling revealed enrichment of type I interferon, inflammatory, and chemokine-related signatures, together with suppression of cell-cycle, translation, and metabolic programs. At the cellular level, 211At injury was accompanied by {gamma}H2AX accumulation, G2/M arrest, increased reactive oxygen species and cytosolic double-stranded DNA, and senescence-like changes. These features arose without consistent increases in classical damage-associated molecular patterns or prominent induction of senescence-associated secretory phenotype factors at 1 MBq/mL. In vivo pharmacologic inhibition of stimulator of interferon genes (STING) signaling attenuated the antimetastatic effect of vaccination. Together, these findings demonstrate that 211At injury can generate an immunostimulatory tumor-cell state characterized by DNA damage and altered inflammatory signaling. STING contributes to the resulting immune protection, highlighting immunogenic tumor-cell conversion as a consideration in the design of targeted alpha therapies.

immunology↗

bioRxiv2026

MetaboXcan: A multiomic framework linking genetically predicted metabolites, gene expression, and complex traits

Genetically regulated molecular phenotypes, such as gene expression and metabolites, are widely thought to mediate the effect of disease-relevant genetic loci identified by genome-wide association studies (GWAS); however, molecular mechanisms connecting genetic variation to disease remain poorly understood. While transcriptome-wide association studies (TWAS) have identified disease-associated genes, strategies that integrate the metabolome remain underexplored. Here, we introduce MetaboXcan, a framework for predicting plasma metabolite levels from genetic data and associating them with complex traits using GWAS summary statistics. We trained lasso regression models on plasma metabolite and genotype data from the Metabolic Syndrome in Men Study (METSIM); these models outperform existing genetic metabolite predictors and generalize well across independent cohorts. MetaboXcan leverages these models to perform four complementary association analyses: (i) gene-to-trait (TWAS), (ii) gene-to-metabolite (M-TWAS), (iii) metabolite-to-trait (MWAS), and (iv) gene-expression-based metabolite-to-trait associations (g-MWAS). The framework further organizes results into metabolic pathways and gene-metabolite interaction networks to facilitate biological interpretation. Applied to chronic kidney disease (CKD), MetaboXcan identified known disease risk genes (PDILT/UMOD, SPATA5L1/GATM), as well as multiple CKD-relevant metabolites (e.g., glycine, homoarginine). Our integrated multiomic analysis enables biological interpretation by revealing glycine availability-centered biochemical axes spanning oxidative stress, cellular energetics, and vascular signaling, while nominating candidate mechanisms for downstream investigation. These results validate known disease-related biology with genetic evidence and generate new hypotheses for further investigation. MetaboXcan is publicly available and broadly applicable to any GWAS phenotype.

genetics↗

bioRxiv2026

Oral Gut Microbial Axis in Inflammatory Bowel Disease and Primary Sclerosing Cholangitis

The oral cavity is increasingly recognized as a reservoir of microbes that can translocate to and influence the gut microbiome. This oral-gut microbial axis may contribute to the pathogenesis of chronic gastrointestinal and hepatobiliary disorders, including inflammatory bowel disease (IBD) and its comorbidity, primary sclerosing cholangitis (PSC). To investigate the oral-gut microbial axis in IBD and PSC, we enrolled 191 participants spanning Crohns disease (CD), ulcerative colitis (UC), CD with PSC (CD-PSC), UC with PSC (UC-PSC), and healthy controls. We generated and analyzed the whole-metagenome shotgun sequencing data from saliva, tongue swabs, and fecal samples. Across oral and gut niches, we identified multiple microbial species differentially enriched in participants with UC compared with healthy controls, including Fusobacterium nucleatum and Gemella sanguinis in saliva, Catonella massiliensis and Tannerella serpentiformis on the tongue, and G. sanguinis and Sellimonas intestinalis in feces. Paired oral-gut analyses revealed 15 potential oral-origin species enriched in fecal samples; notably, G. sanguinis and Veillonella rogosae were consistently enriched in participants with UC. These findings were further supported by an external validation dataset comprising 1,716 gut metagenomes from five independent IBD cohorts, highlighting the reproducibility of oral microbial signatures. In exploratory analyses, smoking history was associated with shifts in the oral microbiome toward a UC-like state in healthy controls, suggesting a possible environmental modifier of the oral-gut microbial axis. Collectively, our study identifies distinct oral microbial signatures and their potential translocation to the gut in IBD and PSC, underscoring the role of the oral- gut microbial axis in disease progression.

microbiology↗

bioRxiv2026

ANCHOR: haplotype-aware allelic and isoform inference from single-cell long-read RNA sequencing with de novo variant calling

Long-read RNA sequencing enables haplotype- and isoform-resolved allelic analysis of transcriptomes, yet extending this capability to single cells and distinct cell types remains computationally challenging due to sparse coverage, sequencing errors, incomplete variant information, and reference-biased transcript assignment. Here we present ANCHOR, a haplotype-aware framework for single-cell long-read RNA sequencing that performs de novo expressed-variant discovery, molecule-level haplotype assignment and isoform-resolved allelic quantification. ANCHOR combines a signed-graph variant caller, pair hidden Markov modelling and beta-binomial UMI aggregation to infer parental allele counts for genes and splice-resolved isoforms, without requiring a pre-existing phased genotype or deep learning. In human single-cell long-read RNA benchmarks, ANCHOR improved variant-calling performance over tested long-read RNA callers at single-cell and low-to-moderate coverage, and its beta-binomial model reduced depth-driven false positives in allele-specific expression testing. Applied to newly generated single-cell long-read RNA-seq data from reciprocal mouse crosses during gastrulation, ANCHOR resolved cell-type- and isoform-specific parent-of-origin imprinting and identified an antagonistic maternally biased Sgce isoform. ANCHOR provides a general framework for allele-and isoform-resolved analysis of diploid single-cell long-read transcriptomes.

bioinformatics↗

bioRxiv2026

Coronavirus envelope protein drives iron sensing disorder by hijacking the TAp73-FDXR axis

Iron overload is increasingly recognized as a critical contributor to coronavirus pathogenesis1, yet the underlying induction mechanisms remain unclear. Here, we uncover a fundamental pathway by which coronavirus drives IRP1 RNA-binding activity to induce iron accumulation2 via targeting the TAp73-FDXR axis. Specifically, coronavirus infection represses transcription of FDXR (encoding the key rate-limiting enzyme in host iron-sulfur cluster synthesis3), thereby impairing host iron-sulfur cluster generation to trigger the functional conversion of the cytosolic aconitase 1 (ACO1) into iron-regulatory protein 1 (IRP1)4, ultimately leading to the hosts persistently false perception of iron deficiency. We identify TAp73 as the primary transcription factor governing FDXR expression, and demonstrate that the coronavirus envelope protein (CoV-E) orchestrates TAp73 nuclear export. Subsequently, CoV-E binds TAp73 through a critical valine residue within its C-terminal PBM domain, inducing the K48-linked ubiquitination and proteasomal degradation of TAp73. Furthermore, we developed a CoV-E-targeting molecule, DPTP-FC, which blocks CoV-E-TAp73 interaction via forming steric hindrance and effectively alleviates iron accumulation and tissue damage caused by PEDV, PDCoV, and SARS-CoV-2 infection. Our study reveals the central role of the TAp73-FDXR axis in CoV-induced iron accumulation, highlighting CoV-E as an attractive antiviral target and DPTP-FC as a promising therapeutic candidate.

microbiology↗

bioRxiv2026

A cerebellar cognitive rheostat bidirectionally controls attention

Attention requires filtering distractors and amplifying signals, processes classically attributed to cortico-thalamic networks. Here, we reveal that the cerebellum operates as a bidirectional "cognitive rheostat" to optimize attentional states. In mice, the anterior and posterior cerebellar vermis exert opposing control over attention. Granule cells in the anterior vermis are functionally suppressed to gate sensorimotor noise via reticular nucleus-driven feedforward inhibition. Conversely, posterior granule cells are recruited by pontine inputs to amplify cognitive signals, a process relying on Grin1-mediated NMDA receptor plasticity. Circuit-specific manipulations targeting this push-pull mechanism, or localized pharmacological modulation, successfully rescued attentional deficits in an ADHD mouse model. These findings fundamentally expand the cerebellums role beyond motor coordination, identifying a topographic circuit algorithm essential for cognitive control.

animal behavior and cognition↗

bioRxiv2026

Dietary depletion of glutamine is atheroprotective

Heart attacks and strokes are late-stage complications of rupture of unstable atherosclerotic plaques. Stable plaques contain stabilizing matrix-producing fibrotic cells, largely smooth muscle cell (SMC)-derived. The molecular drivers of SMC phenotypic transitions to beneficial fibrotic or destabilizing inflammatory and calcifying phenotypes are unclear. Since atherosclerosis develops over decades, there is extensive interest in identifying dietary alterations that enhance plaque stability. We demonstrate that SMC acquire a fibrotic phenotype dependent on glutamine-derived metabolites supporting both catabolism and collagen synthesis. Moreover, dietary glutamine restriction decreases mortality of mice susceptible to atherosclerotic plaque rupture. Lesions from glutamine-restricted mice are smaller and have increased SMC investment. This study identifies dietary glutamine as a driver of cardiovascular mortality, suggesting a new strategy for reducing late-stage complications of atherosclerosis.

physiology↗

bioRxiv2026

Magnetic DNA Origami Nanorotors

Self-assembled DNA nanostructures show great promise as functional devices, highly configurable materials, and in nanorobotics. Magnetic control can provide a powerful actuation mechanism in a broad range of contexts, since it affords a high-level of external control, it is biocompatible, and orthogonal to chemical or electrical stimuli. Here we demonstrate magnetic molecular nanoactuators by leveraging the unique site-specificity of DNA origami to assemble highly anisotropic magnetic nanocubes on high-aspect ratio DNA origami bundles. We traced and controlled 100s of our DNA origami nanorotors at the single-rotor level and demonstrated their programmable magnetic clamping and controlled rotation under uniform and rotating magnetic fields. By varying the population and inter-particle spacing of the nanocubes, magnetic torque values in the order of 10-100 pN nm are achieved at field strengths < 10 mT. Monte Carlo simulations reveal that assembly of nanocubes on DNA origami rotors leads to collective magnetic properties, with numerically estimated torque values in good agreement with the experiments. Our magnetic nanorotors offer a foundation for biocompatible nanorobotics, as well as high-throughput magnetic force and torque tweezers.

biophysics↗

bioRxiv2026

HSC71 acetylation confers protection against Spiroplasma eriocheiris infection by inhibiting apoptosis and promoting ROS production in arthropods

Heat shock proteins (HSP) are increasingly recognized as pivotal regulators of host innate immunity. However, the role of HSP70 post-translational modifications in modulating the immunologic functions during pathogenic infection remains poorly understood. Here, we demonstrated that heat shock cognate 71 kDa protein (HSC71), a member of the HSP70 family, was deacetylated following infection with intracellular pathogen Spiroplasma eriocheiris. HSC71 deficiency exacerbated hemocyte apoptosis and increased host susceptibility to S. eriocheiris infection. Conversely, ectopic expression of hyperacetylation-mimetic HSC71(K579Q) in Drosophila S2 cells conferred robust protection against S. eriocheiris by suppressing apoptosis and augmenting reactive oxygen species (ROS) production. Mechanistically, carnitine O-acetyltransferase (Crat) was identified as the specific acetyltransferase for HSC71. Crat acetylated HSC71 at lysine 579 (K579), which in turn impeded its interaction with the E3 ubiquitin ligase CHIP, thereby stabilizing HSC71 by attenuating ubiquitin-mediated degradation. Crat deficiency induced apoptosis and reduced ROS levels in hemocytes, thereby exacerbating S. eriocheiris intracellular proliferation. Furthermore, acetylation of HSC71 at K579 weakened its binding to superoxide dismutase (SOD), resulting in impaired SOD activity and consequent accumulation of microbicidal ROS. Pharmacological inhibition of the deacetylase SIRT1 with EX-527 enhanced HSC71 acetylation and ROS production, conferring resistance to S. eriocheiris infection in crabs. Notably, EX-527 similarly enhanced the acetylation of Drosophila HSC71 homolog, HSPA8, which in turn impaired its interaction with SOD. This led to elevated ROS levels and restricted intracellular proliferation of S. eriocheiris in S2 cells, demonstrating evolutionary conservation of this mechanism among arthropods. Therefore, this study established the modulation of HSC71 acetylation as a promising strategy to enhance arthropod immunity. Author summaryMembers of the HSP70 family are indispensable components of host innate immunity, and their functions are finely tuned by post-translational modifications. This study investigated the mechanisms that underlie acetylation modification of HSC71 mediated resistance to S. eriocheiris infection in crab. Our results revealed that the acetyltransferase Crat and the antioxidant enzyme SOD directly interacted with HSC71. Mechanistically, Crat acetylated HSC71 at K579, which prevented its ubiquitination by promoting the disassociation of E3 ubiquitin ligase CHIP, thereby improving HSC71 stability. In HSC71-deficient or Crat-deficient crabs, hemocyte apoptosis was markedly enhanced, leading to higher host mortality upon S. eriocheiris challenge. Meanwhile, K579 acetylation on HSC71 weakened the interaction between HSC71 and SOD, resulting in the accumulation of intracellular ROS and thereby restricting S. eriocheiris propagation. Administration of the SIRT1 inhibitor EX-527 enhanced HSC71 acetylation, elevated ROS production and reduced host susceptibility to S. eriocheiris infection. These findings revealed a novel role for Crat-mediated acetylation of HSC71 in orchestrating host defense through two synergistic mechanisms: suppressing apoptosis and promoting ROS accumulation. Thus, targeting HSC71 acetylation may represent a promising avenue to combat S. eriocheiris infection.

immunology↗

bioRxiv2026

Excessive Ca2+-dependent ER-mitochondrial contact stabilization by EFHD1 drives liver injury

Metabolic-associated steatohepatitis (MASH) involves hepatocyte damage that cannot be explained solely by lipid accumulation. Here, to discover injury-specific pathways, we focused on a gene of uncertain function, EF-Hand Domain Family Member D1 (EFHD1), identified in human genome-wide association studies of liver injury but not liver fat. We show that EFHD1, a Ca2+-dependent actin crosslinker, stabilizes endoplasmic reticulum-mitochondria contact sites (ERMCS), detecting spatiotemporal coincidence of inter-organellar proximity and ER Ca2+ release. During MASH, EFHD1 upregulation drives pathological mitochondrial fragmentation via excessive contact persistence. This structural failure promotes mitochondrial double-stranded RNA escape and activation of a maladaptive antiviral PKR-dependent stress response, a causal relationship also supported by Mendelian randomization in humans. Consequently, inhibiting EFHD1 in human and mouse models blunts hepatocyte damage. These findings identify EFHD1 as a Ca2+-dependent ERMCS stabilizer, reveal a hepatocyte-intrinsic injury pathway, and suggest EFHD1 inhibition as a therapeutic strategy.

cell biology↗

bioRxiv2026

EBNA1 INHIBITORS REVEAL CDC7 AND POU2F1 AS DIRECT FUNCTIONAL TARGETS IN EBV EPITHELIAL CANCERS

Epstein-Barr virus (EBV) latent infection is causally linked to several epithelial cancers, including endemic forms of undifferentiated nasopharyngeal carcinoma (NPC) and to a subtype of gastric cancer (GC). EBNA1 is the viral-encoded sequence-specific DNA-binding protein required for episome maintenance but also contributes to host-cell survival through multiple mechanisms including binding to host chromosome. We previously developed small molecule inhibitors of EBNA1 DNA-binding that block host cell cycle progression and growth of EBV+ tumors in vivo. However, the underlying molecular mechanisms of EBNA1 function and inhibition have not been completely elucidated. In this study, we employ VK1727 to inhibit EBNA1 DNA-binding to viral and cellular genomes in three EBV+ epithelial tumors (PDX C15, C666-1 and SNU719). We integrate EBNA1ChIP-seq and transcriptomic RNA-seq analyses to identify the cell cycle dependent kinase CDC7 and a stem cell transcription factor POU2F1 as direct functional targets of EBNA1 in these epithelial cancers. EBNA1 binding to CDC7 promoter and POU2F1 intron promotes RNA Pol II-pS5 to initiate transcription of these two genes. We show that CDC7 inhibitor Simurosertib is epistatic, while Bcl2 inhibitor Venetoclax is synergistic with VK1727 in the inhibition of EBV+ epithelial cancer cell proliferation and survival. Our study reveals new functional gene targets and pathways of VK1727 in EBV+ epithelial cancers that provide new biomarkers and combinatorial strategies to treat EBV-driven cancers. IMPORTANCEEBNA1 is essential for EBV latency and tumorigenesis, but its mechanism of action on host gene expression is not yet known. Small molecule inhibitors of EBNA1 DNA-binding block cell cycle progression and inhibit growth of EBV+ tumors. In this study, we use the EBNA1 small molecule inhibitor VK1727 to identify cellular gene targets that are bound by EBNA1 and deregulated by its pharmacological inhibition in EBV+ epithelial cancer cell lines and an NPC PDX mouse model. We identify cycle dependent kinase CDC7 and the stem cell transcription factor POU2F1 as EBNA1 bound and regulated genes important for EBV epithelial cancer proliferation. These findings not only decipher molecular mechanism how VK1727 blocks cell cycle progression and inhibits cell proliferation but also provide two new cellular gene targets and pathways for therapeutic intervention in EBV+ epithelial cancers.

microbiology↗

bioRxiv2026

A micropeptide component of the OGDH complex modulates ATP production in the TCA cycle

Since its elucidation, the tricarboxylic acid (TCA) cycle has been regarded as a fully defined enzymatic pathway. Here, we report the discovery of a previously unrecognized micropeptide component of this central metabolic system. Through analyzing ribosome profiling data from the mouse sORFs.org database, we discovered APPM (ATP production-promoting micropeptide), a 29-amino acid peptide encoded by a non-canonical small open reading frame within the mouse Ctsb gene. APPM localizes to the mitochondrial matrix and inner membrane, where it directly binds to the E1 subunit (OGDH) of the -ketoglutarate dehydrogenase complex (OGDHc). This interaction modulates OGDHc enzymatic activity, thereby regulating NADH and ATP production. We identified tyrosine-13 as the critical residue mediating the APPM-OGDH interaction and functional activation. Exogenous APPM administration increased ATP levels in vivo and, in a mouse model of heart failure, restored cardiac function, outperforming the reference drug trimetazidine. Collectively, our findings identify APPM as the first micropeptide regulator of the OGDH complex, expanding the known repertoire of the TCA cycle and presenting a potential therapeutic strategy for bioenergetic disorders.

molecular biology↗

bioRxiv2025

A streamlined nanopore-compatible 5PSeq protocol for rapid phenotypic antimicrobial sensitivity testing

Antimicrobial resistance (AMR) poses a significant threat to public health. Rapid and accurate antimicrobial sensitivity testing is essential to guide effective treatment. Here, we present "simplified 5PSeq" (s5PSeq), a streamlined protocol for profiling 5 monophosphorylated (5P) mRNA degradation intermediates that reflect ribosome dynamics in vivo. By capturing antibiotic-induced, context-specific ribosome stalling events, s5PSeq provides a molecular proxy for bacterial growth inhibition--offering a molecular phenotypic readout without the need for culturing. s5PSeq reduces library preparation time to under four hours and incorporates a novel rRNA blocking strategy. We demonstrated its clinical utility by identifying erythromycin-resistant and sensitive Clostridioides difficile clinical isolates. Combining s5PSeq with real-time nanopore sequencing enables fast AMR diagnosis with as few as 3000 reads. In addition to simplifying the study of 5P co-translational mRNA decay, our work suggests that utilizing information-rich phenotypic molecular readouts can significantly improve AMR diagnostics. HighlightsO_LIs5PSeq is a streamlined protocol for profiling 5P mRNA degradation intermediates. C_LIO_LIContext-specific ribosome stalls can be used to assess phenotypic antimicrobial sensitivity at the molecular level. C_LIO_LIBlocking rRNA sequencing at the ligation step streamlines library preparation and lowers costs and hands-on time. C_LIO_LIIntegration with nanopore sequencing allows same-day antimicrobial sensitivity testing in species with 5-3 exonuclease. C_LI

microbiology↗

bioRxiv2025

Task-similarity dependent reconfiguration of compositional modules and geometry in frontal cortex

Task rules critically shape how neural populations encode information and generalize across cognitive demands, yet the mechanisms linking rule structure, neural dynamics, and behavior remain poorly understood. Here we investigate how rule congruence modulates neural-ensemble compositionality, geometry and cross-task generalizability in mouse anterior lateral motor cortex (ALM). Mice switched between tactile and auditory delayed-response tasks where sensory-motor contingencies were either aligned (congruent) or conflicting (incongruent). Recording from 11,000 ALM neurons revealed rule dependent compositional coding of stimulus, choice, and outcome. In contrast to congruent switching, incongruent rules reduced cross-task generalizability of sensory, choice and outcome representations, expanded population dimensionality, increased reliance on nonlinear mixed-selectivity, and produced more complex coding geometry. These neural changes paralleled behavioral costs during task transitions. Moreover, congruence dependent reconfiguration of sensory-choice subspaces in ALM predicted differences in transition costs, implicating ALM in adaptively resolving conflicting contingencies. Together, these results show that task congruence regulates neural coding geometry and population dynamics, linking rule-switching complexity to dimensionality of neural resource allocation and to cognitive flexibility.

neuroscience↗

bioRxiv2025

Temporal dynamics of noradrenaline release at fine spatial scales during motor learning

Noradrenaline (NA) released from the locus coeruleus (LC) has been known to play pivotal roles in arousal, sensory processing, decision-making, and learning through global release across the entire neocortex. Recent studies have demonstrated heterogeneous and modular NA release in distinct brain regions and highlighted its spatiotemporal dynamics across the neocortex. However, the spatiotemporal specificity of NA release at fine scales within a single brain region remains unclear, and it has not been reported whether the release patterns evolve functionally throughout prolonged learning processes such as motor skill acquisition. Here, by employing in vivo two-photon imaging with various genetically encoded NA sensors, we reveal that behavior-induced NA release in the primary motor cortex (M1) is spatially heterogeneous at the scale of local microcircuitry. Over the course of learning, the release pattern is locally refined, achieving consistent spatial precision within M1. Intriguingly, pharmacological manipulations that disrupt the spatial specificity also alter local neurons activity and representations. Furthermore, LC-NA axonal calcium imaging uncovered two distinct temporal activity patterns, in which non-behavior-related rapid axonal activity (sub-second duration events) profoundly affect the temporally precise behavior-induced persistent axonal activity (seconds duration events). Closed-loop optogenetic manipulations that bi-directionally modulate non-behavior-related rapid events directly impacted the learning process. Together, our results provide novel insights into the temporal dynamics of NA release at fine spatial scales within one brain region and underscore the critical role of local NA specificity in regulating circuit plasticity during motor skill acquisition.

neuroscience↗

bioRxiv2025

Novel therapy for gastric cancer peritoneal dissemination using genetically modified dental pulp cells and astatine-211

Peritoneal dissemination of gastric cancer is a terminal stage with limited treatment options and a five-year survival rate below 10%. To develop a more effective treatment, we created a new approach that uses genetically engineered human dental pulp cells (DPCs) that express the sodium/iodide symporter (NIS). These cells (NIS-DPCs) deliver astatine-211 (At-211) directly to tumor sites. When injected intraperitoneally into a mouse model of gastric cancer dissemination, DPCs, which were isolated and expanded, showed strong tumor-homing ability. Chemotaxis mediated by the CXCR4/SDF-1 axis has been reported to play a role in this accumulation. In addition to histological analysis, fluorescent imaging confirmed the selective accumulation of NIS-DPCs within tumor lesions. Introducing the NIS gene markedly increased SLC5A5 expression, enabling efficient At-211 uptake. To avoid nonspecific binding, sodium At-211 was utilized. High-resolution alpha-particle imaging visualized alpha-ray emission specifically from NIS-DPCs, confirming the radionuclides intracellular retention. In vivo, the sequential administration of NIS-DPCs, followed by Na[At-211], resulted in the pronounced regression of peritoneal tumors and a significant extension of survival compared to controls. The therapeutic mechanism involves three coordinated steps: tumor-directed migration of NIS-DPCs, At-211 uptake via NIS transporters, and localized alpha-particle-mediated cytotoxicity. This study introduces the novel concept of cell-based alpha-radiotherapy, which integrates regenerative and nuclear medicine. Due to the well-documented safety profile of DPCs in human clinical trials (J-REPAIR, jRCT1080224505), the NIS-DPC platform emerges as a promising approach for the precise, localized irradiation of disseminated gastric cancer and other challenging malignancies.

cancer biology↗

bioRxiv2025

Whole body MondoA deletion protects against diet-induced obesity through uncontrolled multi-organ substrate utilization and futile cycling

ObjectiveDelineating the nodal control points that maintain whole-body energy homeostasis is critical for understanding potential treatments of obesity and cardiometabolic diseases. The nutrient-sensing transcription factor MondoA is a regulator of skeletal muscle fuel storage, where muscle-specific inhibition improves glucose tolerance and insulin sensitivity. However, the role of MondoA in whole body energy metabolic homeostasis is not understood. MethodsGeneralized MondoA knockout (gKO) mice were generated and assessed for glucose tolerance and insulin sensitivity, body composition, energy expenditure, cold tolerance, and tissue specific transcriptional changes in response to high fat diet. Complementary studies in cultured human adipocytes assessed the impact of MondoA deficiency on substrate utilization and lipolysis. ResultsgKO mice are protected from diet-induced obesity and insulin resistance, through increased whole body energy expenditure. gKO mice exhibit reduced brown and inguinal white adipose tissue mass, without evidence of beiging. The gKO mice are hyperlactatemic and isolated MondoA-deficient adipocytes have increased 2-deoxyglucose uptake and glycolytic function. Lastly, gKO mice and KO adipocytes display increased circulating glycerol relative to free fatty acids in response to adrenergic stimulus consistent with elevated re-esterification. However, this phenotype is not recapitulated in adipocyte-specific KO mice. ConclusionsMondoA deficiency alters cellular sensing of nutrient availability and storage/utilization mechanisms. In the whole-body setting, this results in increased energy expenditure, potentially related to increased glucose uptake and glycolytic flux driving glycerol synthesis to supply high rates of lipolysis and lipid re-esterification. These results suggest that MondoA functions to maintain fuel storage and when lost, inter-organ futile cycling ensues. O_FIG O_LINKSMALLFIG WIDTH=140 HEIGHT=200 SRC="FIGDIR/small/680559v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@1ccf42dorg.highwire.dtl.DTLVardef@b2da28org.highwire.dtl.DTLVardef@1081aforg.highwire.dtl.DTLVardef@1b2168d_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical Abstract.C_FLOATNO 1) Global MondoA deficiency drives 2) tissue glucose uptake which in skeletal muscle is 3) converted and excreted as lactate, while in adipose tissue 4) triglyceride re-esterification requires 5) de novo glycerol synthesis to feed into the futile cycle. C_FIG

physiology↗