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Li, T.

Publications and source records attributed to Li, T..

At least 109 records · Page 6Linked to original sources

CellFluxV2: An Image Generative Foundation Model for Virtual Cell Modeling

Building a virtual cell that simulates cellular behavior in silico is a central goal of computational biology. We introduce CellFluxV2, an image-generative model that predicts how cell morphologies change in response to chemical and genetic perturbations. A core innovation of CellFluxV2 is to learn distribution-level transformations from unperturbed to perturbed cells within the same experimental batch using flow matching, enabling it to disentangle true perturbation effects from confounding batch effects. Incorporating three methodological advances, CellFluxV2 achieves up to a 77% improvement in image fidelity over diffusion- and GAN-based baselines, while maintaining biological fidelity comparable to ground-truth images. Scaling up CellFluxV2, we establish the first scaling laws in image-based virtual cell modeling, demonstrating that performance improves consistently with both dataset size and model capacity. Furthermore, the scaled-up model generalizes well to out-of-distribution perturbations and exhibits two novel capabilities: batch-effect correction and cell-state interpolation. Together, these results position CellFluxV2 as a powerful foundation model advancing the vision of a virtual cell, unlocking novel opportunities for in silico drug screening.

bioinformatics↗

Gn1a rice promotes symbiotic fish growth via reprogramming soil microbiome

Rice-fish symbiotic ecosystems significantly contribute to world food supply. Mutation in rice (Oryza sativa) quantitative trait locus Gn1a, encoding a cytokinin oxidase/dehydrogenase (OsCKX2), leads to cytokinin accumulation in plant, which promotes tillering and panicle branching to achieved higher yield and also shows a selective effect on soil microbial composition. Given that the growth of aquatic animals can be shaped by their surrounding microbial communities, we asked whether adjusting rice cytokinin levels might synergistically boosts both rice yield and fish growth. Herein, we co-cultured common carp (Cyprinus carpio) with ckx2 rice plants to determine the effect of cytokinins on fish. Surprisingly, compared to wild-type plants, the ckx2 plants enhanced nearly 10% fish growth and improved fish antibacterial capacity against Aeromonas hydrophila and Edwardsiella tarda. In fact, the growth-promoting effect on fish requires mediation by the soil microbiota. Mechanistically, ckx2 plants secreted more cytokinins into soil to reprogram the microbial composition, which subsequently translocated to fish gut and modulated the keystone microbes in gut to facilitate short-chain fatty acid production and carbohydrate degradation. These findings provide the first evidence that rice-derived cytokinins exert a cross-kingdom growth-promoting effect on fish, providing a boosting dual-production strategy for advancing rice-fish symbiotic ecosystem.

plant biology↗

The overall and sequence-specific degradation of soil extracellular 16S rRNA genes across China: rates and influential factors

While extracellular DNA persistence substantially influences soil microbiome investigations, its degradation kinetics remain poorly quantified. Here, we developed a primer-labeled DNA approach coupled with microcosm incubation to determine the overall and sequence-specific degradation rates of extracellular DNA amplicon fragments across China. We observed substantial variations in the overall degradation rates of extracellular 16S rRNA gene amplicon fragments among the study sites, with degradation rate constants ranging from 0.05 to 0.16 day-1. The overall degradation rate constants showed significant correlations with soil moisture content, prokaryotic abundance, prokaryotic community profiles, and mean annual precipitation (MAP). The significant influences of moisture content on the overall degradation rates were further verified by a moisture gradient microcosm experiment. The sequence-specific degradation rate constant profiles were additionally correlated with pH, nitrogen content, and mean annual temperature (MAT). Furthermore, propidium monoazide (PMA)-based exclusion of extracellular DNA signals significantly altered soil prokaryotic abundance, richness, and prokaryotic community profiles, and the pool sizes of sequence-specific extracellular 16S rRNA gene amplicon fragments were significantly correlated with their respective degradation rates. This study developed a methodology for determining the overall and sequence-specific degradation rates of extracellular DNA amplicon fragments, highlighting the profound influences of extracellular DNA on soil microbial research and informing the optimization of environmental DNA technologies. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/699460v2_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@b1b3e2org.highwire.dtl.DTLVardef@98f334org.highwire.dtl.DTLVardef@1870650org.highwire.dtl.DTLVardef@1af7105_HPS_FORMAT_FIGEXP M_FIG C_FIG

ecology↗

Lysosomal Aspartylglucosaminidase Couples Glycoprotein Catabolism to Cell-Surface GlycoRNA Production

GlycoRNAs, small non-coding RNAs covalently linked to N-glycans, reveal an unexpected intersection between RNA biology and glycosylation, with potential roles in immunity and cell-cell communication. The origins and biogenesis of their glycans remain unclear. Using CRISPR-Cas9 screening, metabolic glycan labeling and lysosome-targeted proteomics, we reconfirm canonical N-glycosylation enzymes (STT3A/B) and identify components of the mannose-6-phosphate (M6P) lysosomal sorting pathway (GNPTAB, M6PR) as essential contributors. Strikingly, lysosomal aspartylglucosaminidase (AGA) acts as a rate-limiting regulator, linking glycoprotein catabolism to glycoRNA formation. Loss of AGA collapses both abundance and diversity of RNA-linked N-glycans, while pulse-chase tracing shows glycoprotein-derived glycans are repurposed for RNA glycosylation. These findings position lysosomes as active participants in RNA modification and redefine glycoRNA biogenesis as a metabolically integrated process.

cell biology↗

Innate immune sensing via the cGAS-STING pathway restricts extrachromosomal DNA-driven tumorigenesis

Extrachromosomal DNAs (ecDNAs) are circular DNA fragments frequently found in human cancers, where they amplify oncogenes, drive tumor heterogeneity, and promote therapy resistance and poor prognosis. Despite their prevalence, how ecDNAs interact with the immune system remains poorly understood. Here, we show that the cytosolic DNA sensor cGAS detects ecDNA fragments in the cytoplasm and activates the innate immune response. cGAS and STING are frequently silenced in ecDNA+ tumors through promoter hypermethylation. Restoring cGAS or STING in human and murine ecDNA+ cancer cells reactivates innate immune signaling and selectively suppresses ecDNA+ tumor growth in an immunocompetent mouse model. Using two ecDNA biogenesis models, we show that the cGAS-STING pathway restricts de novo ecDNA formation. Together, our findings identify innate immune sensing as a natural barrier to ecDNA-driven oncogenesis and establish cGAS-STING reactivation as a therapeutic strategy for ecDNA+ cancers. HighlightsO_LIThe cGAS-STING pathway is frequently silenced in ecDNA+ tumors C_LIO_LIRestoration of cGAS in ecDNA+ cells activates innate immune responses C_LIO_LIcGAS expression suppresses ecDNA+ tumor growth in vivo C_LIO_LIThe cGAS-STING pathway restricts de novo ecDNA biogenesis C_LI

cancer biology↗

PtdIns(3,5)P2 Is an Endogenous Ligand of STING in Innate Immune Signaling

Cytosolic DNA exposure triggers innate immune responses through cyclic GMP-AMP (cGAMP) synthase (cGAS)1-3. Upon binding to DNA, cGAS is activated to produce cGAMP, which functions as a second messenger that binds to stimulator of interferon genes (STING), an endoplasmic reticulum (ER)-localized signaling adaptor3-5. STING then traffics from the ER to the Golgi, leading to activation of the kinases TBK1 and IKK and subsequent induction of interferons and other cytokines6-10. Here we show that phosphatidylinositol 3,5-bisphosphate [PtdIns(3,5)P2] is an endogenous ligand of STING that functions together with cGAMP to induce STING activation. Proteomics analysis identified a constitutive interaction between STING and PIKfyve, an enzyme that produces PtdIns(3,5)P2 in mammalian cells. Deletion of PIKfyve blocked STING trafficking from the ER and TBK1 activation. In vitro reconstitution revealed a strong and selective effect of PtdIns(3,5)P2 on STING activation. Purified STING bound directly to PtdIns(3,5)P2 in a fluorescence resonance energy transfer (FRET) assay. Consistently, PtdIns(3,5)P2 promoted cGAMP-induced STING oligomerization by binding to a groove between STING dimers as revealed by cryo-EM (Li et al., co-submitted). Similar to PIKfyve depletion, mutation of the PtdIns(3,5)P2-binding residues in STING blocked its trafficking and downstream signaling. These results reveal PtdIns(3,5)P2 as a lipid ligand of STING with essential roles in innate immunity.

biochemistry↗

MutSimABC: A Simulation-Based Approximate Bayesian Computational Framework for Mutation Rate Inference in Long-Lived Trees

Long-lived trees accumulate somatic mutations over centuries, forming genetic mosaics in which branches carry distinct genotypes shaped by meristem development. Elongation dynamics regulate stem cell lineage maintenance, while branching events redistribute mutations, producing genetic patterns that often diverge from the physical tree topology. Tomimoto and Satake (2023) formalized these processes through mechanistic simulations; however, their framework was designed for forward prediction rather than parameter inference. Estimating mutation rates and developmental parameters from observed data remains computationally intractable for likelihood-based methods. We present MutSimABC, an Approximate Bayesian Computation (ABC) framework that extends the Tomimoto & Satake model to enable simulation-based parameter inference. MutSimABC jointly estimates the mutation rate (), elongation parameter (StD), and branching bias ({sigma}) by comparing observed and simulated mutation distributions without requiring explicit likelihood functions. Validation across 169 simulated datasets with known parameters achieved complete recovery of mutation rate and branching bias, and 99.4% recovery for elongation parameters, within 95% highest posterior density (HPD) intervals. Applied to genomic sequencing data from Eucalyptus melliodora, MutSimABC estimated somatic mutation rates ranging from 2.3 x 10-10 to 1 x 10-10 per site per year and inferred partially stochastic meristem dynamics. This framework enables joint inference of mutation and developmental parameters, advancing the mechanistic analysis of somatic evolution in plants, with flexibility that extends to any long-lived organism.

plant biology↗

Screening and structural characterization of a nanobody targeting the thermostable green fluorescent protein

Nanobody-fluorescent protein pairs are powerful tools in imaging, protein purification, and structural biology studies. While thermostable GFP (TGP) offers improved characteristics over conventional GFP, nanobodies that specifically recognize TGP remain relatively limited. Here, we report the screening and identification of a synthetic nanobody, Sb32, that binds TGP with nanomolar affinity using the ribosome technique. The crystal structure of the Sb32-TGP complex, solved at 1.82 [A] resolution, revealed an unusual binding mode in which Complementarity Determining Region 2 (CDR2) provides the major contribution, rather than the typically dominant CDR3. Moreover, the interface is stabilized by an extensive hydration network, which compensates for relatively few direct contacts and may explain the fast association and dissociation kinetics observed for this complex. These findings expand the repertoire of TGP-specific nanobodies and highlight an alternative strategy by which nanobodies achieve high affinity. Sb32 provides a new reagent for TGP-based applications, with potential utility in membrane protein purification and structural studies.

biophysics↗

Incentive valence differentially engages open- and closed-loop basal ganglia circuits during movement initiation

Incentives modulate voluntary movement, yet the circuitry channeling these signals into motor output remains unclear. Classical models emphasize a closed-loop circuit (CLC) linking dorsal putamen (PUTd) with motor cortex, but this pathway is anatomically segregated from affective processing regions. Anatomical and clinical evidence point to an alternative: an open-loop circuit (OLC) from ventral putamen (PUTv) that may route affective signals to motor cortex. Here, we conducted two experiments to test whether a functional OLC exists in humans and whether it is differentially engaged by incentive conditions. First, in 7 T resting-state fMRI (multi-echo), PUTv showed robust functional connectivity with both affective and motor regions, including the cingulate motor area (CMA), even after accounting for PUTd variance. This connectivity pattern supports the plausibility of an independent pathway linking affective basal ganglia regions to the motor cortex. Second, in 3 T task fMRI (incentivized reaching), jackpot (high-reward) and robber (high-loss avoidance) incentive conditions produced distinct behavioral and neural signatures. Jackpot produced a speed-accuracy trade-off, with faster movement initiation but more false starts. Neurally, this coincided with engagement (BOLD responses relevant for initiation speed) being reduced in CLC nodes but not in OLC. Robber, in contrast, eliminated engagement in both OLC and CLC nodes, instead recruiting stopping-related regions (e.g., STN), consistent with an avoidance phenomenology. Together, these findings support a versatile architecture for movement initiation that flexibly engages distinct cortico-subcortical circuits depending on incentive phenomenology, and offer a candidate mechanism through which affective salience and valence modulate voluntary movement. Significance StatementAffective signals profoundly influence movement, yet the mechanisms linking motivationally relevant contexts with motor behavior remain unclear. Combining ultra-high field (7 T) connectomics with task-based (3 T) neuroimaging, we provide the first systems-level evidence in humans for such a mechanism: a ventral putamen-centered open-loop circuit (OLC) connecting affective and motor areas, operating alongside the canonical dorsal putamen-centered closed-loop sensorimotor circuit (CLC). Critically, the phenomenological quality of incentive (how it is construed as reward versus threat) rather than magnitude alone, likely determines which circuit dominates during movement initiation. These findings help to explain paradoxical kinesia in Parkinsons disease, where affective contexts can bypass degraded sensorimotor circuits, and establish foundations for context-based therapeutic interventions.

neuroscience↗

COPS5 is Essential for Sertoli Cell Function and Male Fertility in Mice

The COP9 signalosome subunit 5 (COPS5) is a multifunctional protein that regulates ubiquitin-dependent processes. Global knockout of Cops5 is embryonically lethal, and while it is known to be vital in germ cells and testicular smooth muscle, its function in Sertoli cells, the key somatic supporters of spermatogenesis, remains entirely unknown. This study investigates the critical role of COPS5 within Sertoli cells. Using Sertoli cell-specific Cops5 knockout mouse models, we demonstrate that COPS5 is essential for maintaining male fertility. Sertoli-specific Cops5 ablation resulted in age-dependent male infertility, despite normal initial development. Mutants exhibited progressive testicular atrophy, oligoasthenospermia, and significantly reduced testis weight. Histology showed vacuolated, disorganized tubules devoid of germ cells and sperm. Crucially, COPS5 loss disrupted Sertoli cell polarity, evidenced by aberrant cytoplasmic mislocalization of the nuclear marker WT1 and detachment from the basement membrane. Integrity of the blood-testis barrier (BTB) was severely compromised, with discontinuous and punctate expression of the tight junction adaptor ZO-1. Intercellular communication was also impaired, shown by a stark reduction in Connexin 43 gap junction signals. Our findings establish that COPS5 is an indispensable intrinsic regulator of Sertoli cell function. Its loss disrupts cell polarity, BTB architecture, and gap junction communication, leading to failed support of spermatogenesis and consequently infertility. This work defines a novel and critical somatic function for COPS5 in male reproduction. We conclude that COPS5 is intrinsically required in Sertoli cells to maintain their polarity and BTB function, which are foundational for supporting germ cell development and ensuring male fertility. This identifies COPS5 as a novel, essential regulator within the testicular somatic compartment.

genetics↗

Targeting Putrescine Homeostasis with Lactiplantibacillus plantarum PS128 to Mitigate Autism-like Behaviors in Fmr1 KO Mice

BackgroundAutism spectrum disorder (ASD) is biologically heterogeneous and has limited mechanism-informed interventions targeting core behavioral symptoms. Emerging evidence suggests that gut microbial metabolism shapes neurobehavioral outcomes, yet the specific metabolic pathways linking gut ecology to brain function remain incompletely understood. One candidate pathway is polyamine metabolism, which links microbial amino acid metabolism to host regulation and represents a plausible but underexplored contributor to ASD-related phenotypes. Notably, the psychobiotic Lactiplantibacillus plantarum PS128 has been reported to improve social behaviors in individuals with ASD, although the molecular basis for these effects is unclear. In this study, we used Fragile X mental retardation 1 knockout (Fmr1 KO) mice, a well-established ASD model, to investigate microbiota-dependent metabolic mechanisms underlying autism-like behaviors. ResultsWe found that autism-like behaviors in Fmr1 KO mice were associated with gut dysbiosis, impaired intestinal barrier integrity, disruption of the arginine-ornithine-polyamine pathway, and elevated putrescine in the prefrontal cortex (PFC). Supplementation with PS128 remodeled the gut microbiota, reduced inflammation- and disease-associated taxa, improved intestinal structure and permeability, and restored polyamine homeostasis. Targeted metabolomics revealed an increased PFC-to-serum putrescine ratio in KO mice, which was normalized following PS128 intervention. This correction was accompanied by reduced expression of polyamine transporters in the PFC, including ATP13A family members. Causal experiments supported a functional role for putrescine, as peripheral elevation of putrescine induced autism-like behaviors in wild-type mice, whereas pharmacological inhibition of putrescine synthesis ameliorated behavioral deficits in Fmr1 KO mice. ConclusionsThese findings identify putrescine metabolic dysregulation as a key contributor to autism-like phenotypes and support the existence of an ASD subtype defined by disruption of the arginine-ornithine-polyamine axis. Integrated multi-omics and causal perturbation analyses support a model in which microbiota-targeted intervention rebalances systemic polyamine regulation along the gut-brain axis, thereby improving ASD-relevant behaviors. Our work provides mechanistic evidence linking microbial metabolism to neurochemical homeostasis and highlights the translational potential of metabolic stratification in ASD. Graphical AbstractWe identify dysregulated microbiota-driven polyamine homeostasis as a causal contributor to autism-like behaviors in Fmr1 knockout mice. A psychobiotic intervention restores gut-brain metabolic balance and rescues social and repetitive behavioral deficits through coordinated microbial and host transport regulation. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/694366v2_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@e1e780org.highwire.dtl.DTLVardef@125a3baorg.highwire.dtl.DTLVardef@201228org.highwire.dtl.DTLVardef@1880650_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

Neural geometric representations of social memory for multi-individuals in medial prefrontal cortex

Social memory, an ability to recognize acquainted individuals and related experience, is mainly stored in the medial prefrontal cortex (mPFC). However, it remains unknown how the mPFC encodes the memory of interacted multi-individuals. We, here, report that tuned low-dimensional, geometric subspaces in population neural activity of mPFC neurons encode this form of social memory. Different social experience, in which mice encounter four conspecifics associated with reward, aversive or neutral stimulus, respectively, substantially enhances segregation among identity-specific neural geometric subspaces. Progressive recruitments of mPFC neurons conjunctively encoding social identity and associated social valences underlie the enhanced neural representations of different individuals. Thus, our findings elucidate that tuned neural subspaces in mPFC population activities stably encode and maintain the memory of acquired multi-individuals over experience.

neuroscience↗

Identification of Vector Organisms and Their Carried Pathogens by Metagenomic Sequencing

Vector surveillance is vital for public health, yet traditional methods often face limitations. This study assessed the use of metagenomic sequencing for analyzing mosquitoes, cockroaches, flies, and rodents from Zhongshan, China. The results revealed a high concordance between metagenomic and Sanger sequencing for species identification (Kappa = 0.667, P = 0.011). Metagenomic analysis identified vector-specific dominant bacteria, including Wolbachia in mosquitoes, Blattabacterium sp. in cockroaches, Bartonella tribocorum and Leptospira interrogans in rodents, and Vagococcus teuberi in flies. It also detected numerous classical pathogens, such as Enterococcus faecalis, L. interrogans, and Klebsiella pneumoniae. Among the identified fungi and viruses, 19 were human pathogens, while seven posed a threat to agriculture, notably Pepper mild mottle virus and Tomato brown rugose fruit virus. These findings systematically characterize the microbial communities associated with various vectors, highlight the potential of metagenomics for broad pathogen screening, and underscore its utility in large-scale vector surveillance and public health protection. ImportanceVectors are major carriers of numerous diseases and present risks fot cross-border transmission. Existing detection methods for vectors and their pathogens remain inefficient, insensitive, and prone to difficulties in species identification. Despite extensive efforts in vector identification, the potential threats posed by their associated pathogens remain unclear, demonstrating the need for more effective diagnostic strategies. Metagenomic sequencing offers a promising solution to this challenge. In this study, we applied metagenomic sequencing to vectors collected from urban and port areas in Zhongshan, China, to evaluate its efficacy in pathogen detection. Overall, our work contributes to a better understanding of the public health risks posed by vector-borne pathogens.

microbiology↗

Molecular Surveillance Identifies Evidence of Getah Virus (GETV) in Mosquito Vectors in Alabama, USA

The Getah virus (GETV) is a mosquito-borne RNA virus in the Togaviridae family and the Alphavirus genus, associated with severe disease outbreaks in livestock across Asia-Pacific regions. Since its first isolation in Malaysia in 1955, GETV has been reported in Eurasia and the South Pacific, yet documented cases in the United States remain exceptionally rare, leaving major gaps in regional vector competence and surveillance data. To evaluate the potential of local mosquito populations as GETV carriers, field collections were conducted in Montgomery and Pike Road neighborhoods of Alabama from 2023 to 2024, capturing multiple mosquito species for molecular screening. Initial real-time qPCR assays on 200 pooled and individual samples suggested that approximate 30% of specimens demonstrated possible GETV carriage potential. To validate viral presence, two distinct primer sets were designed to amplify viral genome fragments. Mosquito RNA was reverse-transcribed into cDNA, followed by conventional PCR. The first PCR, targeting [~]280 bp, produced single or multiple amplicon bands in 32% of samples via gel electrophoresis. These positive cDNAs were re-amplified with a second primer pair targeting a [~]430 bp fragment from a separate genomic region, yielding confirmatory bands in 30% of specimens. Amplified products were purified and Sanger sequenced, revealing approximate 95% nucleotide similarity to the wild-type GETV reference genome. This study delivers the first field-based molecular evidence supporting GETV detection in Alabama mosquitoes, signaling either localized emergence or potential introduction of the virus. These findings underscore the need for expanded vector competence profiling, arboviral surveillance, and livestock disease preparedness in the southeastern United States, strengthening both public health readiness and state-level vector monitoring strategies.

microbiology↗

VQ-MAP: a flexible, versatile framework for animal behavior understanding

The brain generates behaviors that reflect configurations and computations of neural systems. Animal pose analysis can comprehensively profile behaviors in terms of stereotyped action expression, but the absence of common tracking standards makes it difficult to relate measurements across labs and experiments, hindering data exchange and collective knowledge building. Here, we present VQ-MAP, a versatile approach capable of mapping 2D or 3D keypoint sequences with divergent pose formats into a common set of discrete behavioral motifs. VQ-MAP utilizes permutation-invariant attention blocks for keypoint-agnostic processing and a quantized latent space for fast, interpretable delineation of motifs organized into type-subtype hierarchies. We deployed VQ-MAP to align and harmonize heterogeneous pose datasets, co-profiling rodent behaviors in multiple species and models of autism. In longitudinal recordings of rat behavioral maturation, VQ-MAP revealed developmental trajectories connecting specific behaviors across periods of substantial bodily growth and change. By unifying heterogeneous behavioral measurements, our framework will accelerate neurobiological discovery and help the community exercise the full potential of large-scale behavioral datasets.

neuroscience↗

Hepatocyte and adipocyte CDO1-mediated intracellular cysteine catabolism differentially modulates diet-induced obesity and fatty liver in mice

Background and AimsPlasma cysteine has been positively linked to BMI in humans. Consistently, dietary cysteine restriction significantly decreases obesity in mice via mechanisms that are incompletely defined. Cysteine dioxygenase type 1 (CDO1) mediates the major cysteine catabolism pathway and is highly expressed in hepatocytes and adipocytes. The goal of this study is to determine the impact of this endogenous cysteine catabolism pathway on obesity and fatty liver disease. MethodsDiet-induced obesity and fatty liver disease were studied in hepatocyte-specific CDO1 knockout mice (L-CDO1-KO), hepatocyte-specific CDO1 transgenic mice (L-CDO1-Tg), adipocyte-specific CDO1 knockout mice (Ad-CDO1-KO), and adipocyte-specific CDO1 transgenic mice (Ad-CDO1-Tg). ResultsDeletion of hepatocyte CDO1 decreased cysteine conversion to cysteine sulfinic acid, and had modest impact on liver cysteine, GSH, or taurine abundance. When fed a Western diet (WD), L-CDO1-KO mice showed elevated liver injury markers and inflammatory infiltration independent of obesity or steatosis. When fed a fibrogenic high fat/cholesterol/fructose diet (HFCFr), L-CDO1-KO mice developed worsened liver fibrosis. In contrast, L-CDO1-Tg mice fed WD showed lower blood glucose, but similar degree of obesity and steatosis compared to WT mice. Deletion of CDO1 in white and brown adipocytes of Ad-CDO1-KO mice had no effect on WD-induced obesity. In contrast, overexpression of CDO1 in white and brown adipocytes attenuated WD-induced obesity, which resulted in reduced hepatic steatosis. ConclusionGenetic manipulation of intracellular cysteine catabolism in hepatocytes and adipocytes differentially modulates diet-induced obesity and fatty liver, which warrants future mechanistic investigation to better understand cellular cysteine sensing mechanisms and cysteine control of cell metabolism.

physiology↗

AAV vectors with enhancer-controlled ITR promoters

Adeno-associated virus (AAV) is limited by its packaging capacity and the unwanted promoter activity of inverted terminal repeats (ITRs). Here we utilized mini-enhancers to regulate ITR promoters and drive AAV cargo expression in the absence of the canonical promoters, which not only solved the ITR issue but also released more payload for the cargo. As an example, this new design successfully enabled robust, tissue-specific SpCas9 gene editing via a single AAV, which otherwise requires a dual-AAV system due to the large size of SpCas9.

bioengineering↗

Alphaviral capsid proteins inhibit stress granule assembly via competitive RNA binding with G3BP1

Viral infection is one of the conditions that induces stress granule (SG) formation, a cellular defense mechanism that exerts antiviral effects. To counteract this host response, viruses have evolved a broad spectrum of strategies to inhibit SG formation. However, the molecular mechanisms underlying SG inhibition remain poorly understood. The nucleocapsid proteins play a critical role in virus replication and host interaction. Here, using Semliki Forest Virus (SFV) as a model, we uncover the function of the alphavirus nucleocapsid in SG inhibition. This inhibitory function depends on oligomerization mediated by an N-terminal -helix and with a positively charged intrinsically disordered region (IDR). We show that SFV capsid directly competes with G3BP1 for RNA binding, thereby disrupting G3BP1-RNA liquid-liquid phase separation (LLPS) in vitro and SG assembly in cells. This mechanism is conserved across the alphavirus family but is not shared by the nucleocapsid of SARS-CoV-2 or other endemic viruses examined. Notably, expression of a peptide from SFV capsid is sufficient to inhibit SG formation induced by Amyotrophic Lateral Sclerosis (ALS)-associated mutations, suggesting potential therapeutic applications. Our findings reveal mechanistic insight into SG modulation by the viral capsid protein and provide a possible bioengineering tool for probing SG dynamics in health and disease.

cell biology↗