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Biology subjects

Fu, T.

Publications and source records attributed to Fu, T..

At least 19 recordsLinked to original sources

Cell-Lens: Advancing LLM Reasoning at Single-Cell Resolution

Recent work applies large language models (LLMs) to single-cell data, but the cell usually reaches the model as a short ranked list of genes. This list drops much of what defines a cell, so the model reasons from a partial view. Yet the information is not lost in the measurement, only in the text. Here, we introduce Cell-Lens, a training-free structured representation of the measured cell. It writes the cell as typed blocks, from paired measurements to marker-supported programs. On CellVerse, SOAR, CellPuzzles, and SC-Arena, across five model families, Cell-Lens improves performance by up to 27.1 points. It also narrows the model gap and reduces API-reported reasoning-token usage. The largest gains occur when a defining RNA marker falls out of the ranked list. In these cases, the paired protein data still capture the corresponding signal. Shuffling the added blocks removes the gain. The gain persists after selected label-associated proteins are removed. These controls link the improvement to cell-matched biological content. We also build and release CellSpectrum, a broader benchmark spanning eight sources and seven tasks with paired modalities. The gains hold there across tasks, tissues, and modalities. These results show that a better cell representation can improve single-cell reasoning without retraining the model. Code and CellSpectrum are publicly available at https://github.com/LiZaiyuan0619/Cell-Lens.

bioinformatics↗

The mitochondrial RNA extrusion-induced innate immunity is regulated by N6-methyladenosine machinery

Mitochondrial RNA (mtRNA) released into the cytosol functions as a damage associated molecular pattern that activates pattern-recognition receptor (PRR)-mediated inflammation, yet its release mechanisms and cytoplasmic fate remain poorly understood. Here we report that chemical Abt-373-treatment and Vesicular stomatitis virus (VSV) infection induce mtRNA extrusion through Bax/Bak and VDAC1 channels, accompanied by mtDNA release. Extruded mtRNA in A549 cells activates multiple cytosolic PRRs, including RIG-I, MDA5, TLR3/7/8, and PKR, each contributing differentially to the innate immune signaling. Analysis of GEO datasets and methylated RNA immunoprecipitation (MeRIP) assays further reveals that mtRNA carries methyladenosine (m6A) modification. m6A machinery proteins are involved in the cytoplasmic retention time of mtRNA and its interaction with RIG-I, thereby modulating mtRNA-induced innate immunity. Thus, our work establishes in vitro models of mtRNA extrusion, and highlights m6A-dependent modulation as a potential therapeutic target for mtRNA-driven inflammation.

immunology↗

Single shot low-dose radiation durably and focally increases cortical excitability: a potential therapy for neuronal circuit disorders?

Herein, we assess the potential of low-dose stereotactic radiosurgery (SRS) to modulate neuronal network states without apparent damage to cellular integrity. Using a small animal radiation research platform (SARRP), a 1 mm3 focal target in the mouse visual cortex was irradiated with doses of 5, 20, and 40 Gy. One-month later a significant dose-dependent increase in excitatory synapse numbers was observed, notably limited to the treated visual cortex and not the adjacent somatosensory cortex. Six months post-irradiation, cortical neuronal microcircuit activity was monitored in awake mice using high sensitivity two-photon calcium imaging. A single 5 Gy dose resulted in a significant microcircuit-wide increase of spontaneous neuronal activity, consistent with a lasting shift in the functional architecture of the irradiated nodal network. At higher SRS doses (40 Gy) this neuromodulatory window appears to close. In aggregate, these data suggest that low-dose radiation could, in some circumstances, be exploited by selected high precision SRS technologies to durably modulate neuronal circuit disorders. Some, or even all the clinical benefits reported in the companion article by Zhao et al. are likely attributable to the biological properties we sought to characterize in our research.

neuroscience↗

Structural Mechanisms of DNAJC13 Dimeric Assembly and InsP6 binding in Recycling Endosome Regulation

The balance of plasma membrane protein degradation and recycling during endocytosis is regulated, in part, by the large J-domain-containing protein DNAJC13/RME-8 and WASH complexes, which function together with Retromer to support cargo trafficking into recycling endosomes. Despite extensive cellular and biochemical studies, the structural basis and proteomic landscape of DNAJC13 function remain elusive. Here, we find that DNAJC13 forms an unexpected antiparallel homodimeric architecture involving distinctive symmetrical interactions between composite IWN1 and -solenoid ARM2 domains in each protomer. Additionally, two PH-like domains of unknown function (PHL2 and PHL3), adjacent to the PI(3)P-binding PHL1 domain, form an unanticipated composite, positively charged pocket occupied by InsP6, as visualized structurally and verified by mass spectrometry. Proteomic profiling of DNAJC13-associated endosomes revealed enrichment of recycling endosomal components and WASH complexes. Mutations disrupting the dimer interface disable recruitment of WASH complexes to endosomes and result in elongated endosomal tubulation. Disruption of InsP6 binding impairs DNAJC13 binding to PI(3)P-containing vesicles in vitro and Transferrin-positive endosomes in cells. We further demonstrate that DNAJC13 dimerization and InsP6 binding promote melanin production during melanosome maturation, a process known to require recycling endosomes. This work provides a structural and mechanistic framework for understanding DNAJC13 function in recycling endosome control.

cell biology↗

ERK signaling integrates multiple mechanisms to drive cortical evolutionary expansion

A critical first step in human evolutionary divergence from chimpanzees and other primates is the production of greatly increased numbers of cortical pyramidal neurons during development and over evolutionary time. This expansion underpins the enlarged human cerebral cortex and, consequently, higher-order cognition and consciousness. However, the cellular and molecular mechanisms driving this progressive increase in cortical neuron number remain incompletely understood. Here we show that elevated ERK signaling in human cortical radial glia (RGs), relative to mouse RGs, arises from evolutionary changes in the developmental expression of existing shared genes. Using the Emx1-Cre line to overexpress MEK1DD, a constitutively active mutant of rat MAP2K1, we found that enhanced ERK signaling in mouse cortical RGs upregulates cAMP-PKA signaling, along with promoting self-renewal, expanding the RG pool, and accelerating cell cycle progression. Conversely, overexpression of PKA signaling in mouse cortical RGs via in utero electroporation of constitutively active PRKACA mutants (L206R or W196G) reduces ERK activity, prolongs the cell cycle, suppresses SHH-SMO and YAP/TAZ pathway activity, and blocks ependymal gliogenesis. Together, our results demonstrate that ERK and PKA signaling in cortical RGs engage a dynamic balance of synergistic and antagonistic interactions. This interplay couples evolutionarily enhanced ERK and PKA activities to increased cortical RG numbers, prolonged cell cycle progression, and an extended neurogenic period, which collectively culminate in a marked increase in neuronal production. We conclude that heightened ERK pathway activity in cortical RGs serves as a central driver of progressive neocortical expansion across mammalian evolution. Significance StatementThe human brains extraordinary cognitive abilities stem from a dramatic increase in cortical neuron numbers during evolution. This study reveals that enhanced ERK signaling in human cortical radial glia neural stem cells, relative to other primates, is a key driver of this expansion. By manipulating this pathway in mice, we demonstrate that ERK coordinates multiple mechanisms--including self-renewal, cell cycle acceleration, and a dynamic interplay with PKA signaling--to boost neuronal output. These findings identify a core molecular axis underlying mammalian cortical evolution and provide a framework for understanding how the human cortex acquired its unique size and, consequently, its remarkable complexity.

developmental biology↗

Performance-impacting brain state maladaptation driving disease progression in early mouse and human neuroinflammation

The neuronal mechanisms driving progression in neuroinflammatory disorders from early relapse-remitting phases to later neurodegenerative phases remain largely elusive. Functional brain state shifts towards hyperactivity, persisting beyond relapses, represent an early maladaptive response. Here, in remission stage of an experimental autoimmune encephalitis (EAE) mouse model of RRMS, we identified a reduced excitability upon optogenetic stimulation in the brain stem, the area of active disease, while in the cortex a persistent cortical neuronal hyperactivity and synaptic remodeling emerged, accompanied with an increase of markers of early apoptosis. In contrast, hippocampal circuits, which undergo a functional state shift without hyperactivity, do not show increased apoptosis. Visual cortical networks showed a deterioration of the accuracy of encoding visual information and a decrease in the behavioural visual discrimination ability in mice. In RRMS patients in remission, we identified a reduced visual colour discrimination, indicating both the presence and the clinical relevance of early brain state maladaptation that may contribute to progression independent from relapse activity (PIRA). SummaryIn a RRMS model and in patients, impaired visual processing was reported, indicating brain state maladaptations, associated with persistent cortical hyperactivity, brain stem hypoactivity, synaptic remodeling, and apoptosis. These maladaptations might contribute to relapse-independent disease progression through sustained network dysfunction.

neuroscience↗

The m6A Landscapes on the RNA of Mumps Virus and the Host 3 Modulate the Viral Replication and Antiviral Innate Immunity

N6-methyladenosine (m6A) is the most prevalent internal modification in eukaryotic mRNA and has emerged as a critical regulator of RNA virus infection. However, its role in the mumps virus (MuV), a non-segmented negative-strand (NNS) RNA virus, remains undefined. Here, we comprehensively characterize the m6A epitranscriptome of MuV JL2 strain and its functional impact on viral replication and host innate immunity. Using single-base resolution GLORI-seq, we identified abundant m6A modifications on MuV genomic, antigenomic and messenger RNAs, with uneven distribution and non-canonical motif enrichment. Genetic depletion of METTL3 enhanced viral replication by facilitating RNA synthesis and nucleocapsid encapsidation in Vero-E6 cells. In epithelial A549 and monocytoid THP1-iDC cells, MuV infection triggered robust type I/III interferon and proinflammatory responses by activating multiple pattern recognition receptors in a m6A-dependent manner and induced incomplete iDC maturation characterized by downregulated HLA class II; however, the latter molecule can be partially restored by METTL3-knockdown. The MuV infection reshaped the host m6A landscape with a positive correlation between m6A enrichment and transcripts enhancement of numerous innate immune genes in a cell type-specific manner. Functional analyses revealed that host m6A machinery modulates antiviral signaling, which varies between viral infection and viral RNA-transfection due to the infection-induced host m6A machinery fluctuation. Collectively, our findings demonstrate that m6A serves as a bidirectional regulator during MuV infection, simultaneously constraining viral replication and modulating host immune recognition, thereby highlighting RNA methylation as a pivotal determinant of MuV pathogenesis and a potential target for optimized immunogenicity. ImportanceN6-methyladenosine (m6A) modification has emerged as a key regulator of RNA virus infection, yet its role in mumps virus (MuV) remains undefined. Here we map the single-base resolution m6A landscape of a recombinant MuV and demonstrate that viral m6A restricts genome encapsidation and replication while attenuating innate immune sensing. MuV infection extensively remodels host m6A profiles in epithelial and dendritic cells, preferentially enhancing methylation of innate immune transcripts. Strikingly, m6A-deficient MuV RNA exhibits increased affinity for RIG-I-like and Toll-like receptors, eliciting stronger interferon and proinflammatory responses. Our findings identify m6A as a bidirectional regulator of MuV replication and immunity and suggest an epitranscriptomic strategy to optimize live-attenuated vaccine design.

immunology↗

Multiple ecological and evolutionary mechanisms drive treatment-induced antibiotic resistance

The emergence of resistance within patients during antibiotic treatment is an important cause of treatment failure. However, the ecological and evolutionary mechanisms driving within-patient emergence remain poorly understood. Here, we analysed 24,478 Pseudomonas aeruginosa isolates sampled from 180 bronchiectasis patients during a clinical trial to understand how ciprofloxacin-resistant infections emerged over a one-year period of pulse-dosing. Pre-existing resistance predominated, accelerating resistance emergence relative to patients where resistance emerged by spontaneous mutation or strain immigration. Selective sweeps of costly mutations increased resistance over time in some patients, whereas in others oscillating resistance levels were driven by antibiotic treatment and resistance-growth trade-offs between genetically divergent subpopulations. Our findings show that infections under identical treatment follow diverse and sometimes complex ecological and evolutionary paths to antibiotic resistance, with implications for better predicting and managing treatment-induced antibiotic resistance.

evolutionary biology↗

Mutant FGFR3 restricts bone yet expands cortex via ERK-mediated self-repression

FGFR3 gain-of-function mutations cause achondroplasia 1,2, the most common form of dwarfism, yet trigger paradoxical cerebral overgrowth and skeletal stunting 3-9. Here, we demonstrate that Fgfr3-K644E mutant drives enhanced ERK activation, which then represses Fgfr3 expression and downregulates ERK signaling. In the developing cortex, this mechanism transiently expands cortical stem cells by activating ERK, then shifts signaling to downregulate Fgfr3/ERK/PKA and enhance YAP/TAZ, promoting cerebral overgrowth and premature ependymal maturation. In growing bones, Fgfr3-K644E mutant first elevates ERK to inhibit chondrocyte pre-hypertrophy, then downregulates Fgfr3/ERK/PKA and upregulates YAP/TAZ to accelerate premature hypertrophy and ossification. Our findings establish that FGFR3 mutations limit bone yet expand cortical size via an ERK negative feedback loop, revealing a unified disease mechanism and novel therapeutic targets.

developmental biology↗

Cell-Type-Resolved Isoform Atlas of Human Tissues Reveals Age and Alzheimer's Disease-Associated Splicing Changes

Alternative splicing is a key mechanism for transcriptomic diversity, but how isoforms map to specific cell types in bulk tissues remains unclear. We present Sciege, a multimodal method that integrates bulk short-read RNA-seq with single-cell and long-read data to estimate cell type-specific isoform distributions. Through simulations, we demonstrate that Sciege accurately estimates isoform abundances and identifies differentially abundant transcripts through statistical tests. Applied to seven tissues in GTEx and brain tissue in ROSMAP datasets, Sciege generates a first-to-date multi-tissue isoform atlas and reveals isoform changes linked to cell types, aging, and Alzheimers disease. Validation with external cohorts and experimental data confirms our findings. Notably, we identify upregulation of the MAPT-010 isoform in AD inhibitory neurons, consistent with known methylation signatures. Our approach demonstrates the value of integrating RNA-seq data to study cell type-specific splicing and provides a foundation for further genetic and functional studies of alternative splicing across biological contexts.

bioinformatics↗

Zwitterionic hydrogel designs for conducting polymers enable bioelectronics with suppressed foreign body response

For long-term, continuous operation of implantable biosensors and electrophysiological devices, the foreign body response (FBR) is a major obstacle that needs to be overcome. As the FBR progresses, any implanted device will become damaged and isolated from its physiological environment, due to encapsulation by fibrotic tissue and inflammatory immune cells. To achieve more compatible and low-impedance biointerfaces, conducting polymers, such as PEDOT:PSS, have been extensively explored as ideal materials. However, FBR on such conducting polymers remains an unmet challenge. We report a zwitteronic-hydrogel-based double-network design for PEDOT:PSS that can significantly suppress the FBR by 64%, in addition to improving conductivity by more than one order of magnitude. Surprisingly, the FBR level of this design is even lower than that of the parent zwitteronic hydrogel by 53%. Our further immunological investigations at the histological, cellular, and transcriptomic levels give deeper insights into the unique effects that come from the chemical heterogeneity. Furthermore, chronic electrocardiographic recording in mice demonstrate the benefit of this material design to long-term, implanted electrophysiology, which provides indications for the future development of immunocompatible electronic polymers.

bioengineering↗

BioDH: A reversible data hiding framework for secure hiding sensitive data in biological data

With the rapid advancement of single-cell RNA sequencing (scRNA-seq) technologies, the open sharing of massive high-dimensional biological data has become a key driver in advancing precision medicine and fundamental research. However, while promoting data accessibility, ensuring effective protection of sensitive data has emerged as a critical challenge in bioinformatics. To address the issue, we propose BioDH (Bi-ological Data Hiding) framework, a reversible information hiding frame-work tailored for biological data, aiming to simultaneously ensure data security and preserve scientific utility. The framework employs optimal lossless compression and strictly controls numerical perturbations to maintain precision. Upon extraction, both the secret information and the original data are perfectly recovered, achieving true reversibility. Validation on real-world datasets shows exceptional fidelity: when embedding capacity is 1.25 bit per byte, maximum and mean absolute errors are 8.54E-04 and 8.90E-11, respectively; PCA reveals a correlation of 1.0; UMAP testing exhibits no structural distortion; Mantel tests and clustering analyses (ARI = 1.0) confirm preservation of high-dimensional topology and cell subpopulations. All metrics surpass biological compatibility thresholds, indicating no detectable impact on downstream analyses.

bioinformatics↗

Plasmid dependent phage eliminate pathogenic bacteria and antibiotic resistance plasmids from the chicken gut microbiome

Conjugative plasmids are a key reservoir of antimicrobial resistance (AMR) in commensal and pathogenic bacteria within the gut microbiome. Plasmid-dependent phage (PDPs) are a promising therapeutic option directly targeting AMR plasmids, but the efficacy of PDP therapy in vivo is poorly understood. Here, we show that treatment with the PRD1 phage eliminated Salmonella enterica carrying a broad-host range IncP AMR plasmid (RP4) from the chicken gut microbiome. Moreover, PRD1 treatment prevented the transfer of the RP4 plasmid to other members of the gut microbiome, whilst causing only minimal disruption to microbiome composition. Our results demonstrate that PDP therapy rapidly eliminates AMR from a natural gut microbiome by killing pathogenic bacteria and blocking AMR plasmid transmission to commensals, highlighting the conjugative pilus as a promising antimicrobial target.

microbiology↗

Sex differences in cytokine induction by activated T cells from hypertensive BPH/2 and normotensive BPN/3 mice

Over the past two decades, considerable evidence has emerged to implicate a role for the immune system in the development of hypertension. Previous studies have shown immune cells contribute to the development of hypertension in multiple animal models, however the role of the immune system in spontaneously hypertensive BPH/2 mice is not clear. In the current studies, found T cells derived from male hypertensive BPH/2 mice demonstrated an attenuated activation as compared to those derived from male BPN/3 normotensive mice. However, we also observed striking sex differences in T cell cytokine production in these strains. At 24 h post activation, in comparison to male BPH/2 mice, activated T cells from male BPN/3 mice secreted more IL-2, IL-3, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-22 and TNF. In contrast to male mice, less than half of these cytokines were different between strains in female mice. We also noted marked differences in early Th17 cytokine production in which IL-17A, IL-17F and IL-22 were greater in the male, but not female, BPN/3 groups. Taken together, the data suggest that polyclonally activated T cells from male, and to a much lesser extent, female BPH/2 mice have a weaker cytokine response as compared to T cells from BPN/3 mice which may be due to an overall attenuated activation of T cells from male BPH/2 mice. Overall, while there are striking differences in T cell response between the BPH2 and BPN/3 strains in male mice, the data indicate far fewer differences between the strains in female mice.

immunology↗

Programmable Aptamer-Embedded Circular RNAs for Targeted Antigen-Presenting Cells Immunotherapy

The targeted delivery of mRNA to specific cell types in vivo remains a major unmet need in mRNA-based therapeutics. Conventional strategies rely on nanocarriers engineered with various modifications. However, these exogenous materials could cause adverse reactions, and their complex manufacturing processes often hinder clinical application. By leveraging the innate programmability of RNA sequences, we herein report a carrier-free, scalable, and programmable aptamer-embedded circular RNAs (Apt-circRNAs) design that confers targeting capability through the strategic incorporation of different aptamers at varying densities, while retaining circular RNA stability and protein production capability. Experimental results show that antigen-loaded Apt-circRNAs enable targeted delivery to antigen-presenting cells (APCs), drive endogenous antigen expression, activate antigen-specific T cells, and elicit potent immune responses that mediate clearance of both early-and late-stage tumors in mouse models. In humanized models of colorectal cancer, the combination of Apt-circRNA and PD-1 blockade produced synergistic antitumor effects and significantly reshaped the tumor microenvironment. In a first-in-human (FIH) clinical trial, Apt-circRNA vaccine encoding KRAS G12D/G12V neoantigens exhibited a favorable safety profile and induced robust immune activation, T cell mobilization, and neoantigen-specific immune responses. With this study, we have established a new paradigm in carrier-free, APC-targeted RNA vaccine design, offering a safe and effective platform for next-generation cancer immunotherapy.

bioengineering↗

ToxiTaRGET: a multi-omics resource for toxicant-responsive molecular targets

Environmental toxicant exposures can induce widespread alterations in both the transcriptome and epigenome of mammals, and directly contribute to the increased risk of various diseases, including cardiovascular disorders, cancer, and neurological disorders. To evaluate how early-life toxicants produce long-term impacts on the transcriptome and epigenome in mice, the Toxicant Exposures and Responses by Genomic and Epigenomic Regulators of Transcription II (TaRGET II) Consortium generated a landmark resource comprising 3,607 multi-omics from longitudinal studies in mice. The molecular changes in responding to distinct environmental toxicants, including arsenic (As), lead (Pb), bisphenol A (BPA), tributyltin (TBT), di-2-ethylhexyl phthalate (DEHP), dioxin (TCDD), and fine particulate matter (PM2.5), were systematically identified and visualized on an integrative platform, ToxiTaRGET, to allow quickly search and browse by researchers. ToxiTaRGET houses a rich repository of molecular signatures, including gene expression, chromatin accessibility, and DNA methylation profiles, in response to early-life toxicant exposures. These molecular signatures span multiple biologically important tissues in both male and female mice at three distinct life stages, offering a valuable resource for the environmental health and toxicogenomic research communities.

bioinformatics↗

ERK and PKA signaling drive the evolutionary expansion of the cortex

The human cerebral cortex, essential for intelligence, cognition, and language, evolved its uniqueness through mechanisms that drive neuronal expansion. Here, we demonstrate that ERK and PKA signaling cooperatively preserve the neurogenic capacity of cortical radial glia (RGs) by inhibiting gliogenic YAP/TAZ and SHH signaling. YAP/TAZ signaling drives cortical RGs toward ependymal fate, while SHH signaling facilitates the generation of cortical tripotential intermediate progenitor cells that produce cortical astrocytes, oligodendrocytes, and olfactory bulb interneurons. During evolution, cortical outer radial glia (oRGs) acquire dominant ERK/PKA signaling through a self-reinforcing feedback mechanism that suppresses both YAP and SHH pathways, thereby significantly enhancing oRG self-renewal capacity and extending the neurogenic period. These findings reveal that cortical neurogenesis, gliogenesis, and evolutionary expansion are coordinately regulated by an integrated ERK, PKA, YAP/TAZ, and SHH signaling, identifying a unifying principle of mammalian cortical development and evolution.

developmental biology↗

Cytosine-to-uracil RNA editing is upregulated by pro-inflammatory stimulation of myeloid cells

Myeloid cells undergo large changes to their gene expression profile in response to inflammatory stimulation. This includes an increase in post-transcriptional modifications carried out by adenosine-to-inosine (A-to-I) and cytosine-to-uracil (C-to-U) RNA editing enzymes. However, the precise RNA editing targets altered by stimulation and the consequences of RNA editing on gene expression and the proteome have been understudied. We present a comprehensive RNA editing analysis of stimulated myeloid cells across three independent cohorts totalling 297 samples, including monocytes and IPS-derived microglia. We observed that C-to-U editing, while less abundant, has a higher effect size in response to stimulation than A-to-I, and has a greater potential to recode the proteome. We investigated the consequences of RNA editing on RNA stability and gene expression using in silico and in vitro reporter methods, and identified a recoding C-to-U site in ARSB that mimics a reported lysosomal storage disorder mutation.

genomics↗