Biology subjects

Luo, W.

Publications and source records attributed to Luo, W..

At least 37 records · Page 2Linked to original sources

bioRxiv2026

Interactome mapping in human excitatory neurons reveals novel risk genes and pathways in Alzheimer's disease

Alzheimers disease (AD) is an irreversible neurodegenerative disease defined by its molecular hallmarks - amyloid beta peptide plaques and neurofibrillary Tau tangles. Despite significant progress that has been made in uncovering a large number of genetic risk factors through extensive genomic sequencing and genetic studies, the molecular mechanisms driving AD-associated pathology and cognitive decline remain poorly understood. Therefore, alongside the identification of more risk genes, it is also paramount to study how these genes function and influence each other within the cellular pathways and overall molecular networks in AD-relevant brain cell types. However, current human protein-protein interactome datasets were all generated in either yeast or generic human cell lines. Consequently, many important neuronal interactions, especially neuron-specific ones, have yet been discovered. To address this critical gap, we developed a highly scalable, high-quality interactome mapping pipeline in human excitatory neurons derived from induced pluripotent stem cells (iPSC), and generated a comprehensive, neuron-specific interactome map, named ADNeuronNet, for key AD risk genes. ADNeuronNet consists of 1,767 high-confidence interactions among 1,189 proteins and is the only dataset enriched with neuron-specific genes when compared to known protein interactions, including previous large-scale interactome maps, for the same baits in the literature. Within ADNeuronNet, we identified 1,375 novel interactions, many of which are likely neuron specific. For example, we identified a neuron-specific interactor, RIN2, for major AD risk factor BIN1 and confirmed RIN2s function in recruiting BIN1 to RAB5 positive early endosomes, a process that has been well-associated with AD etiology. Additionally, we performed quantitative interaction perturbation analyses on AD risk genes with AD-associated mutations or isoforms and identified significant changes in 99 protein interactions among 11 different protein variants. Finally, we found that subunits from the anaphase-promoting complex/cyclosome (APC/C), another novel BIN1 interactors identified by ADNeuronNet, mediated modulation of Tau-aggregation in neurons via regulation of APOE expression, uncovering a previously unrecognized BIN1-APC/C-APOE regulatory axis in AD pathobiology. In summary, these findings illustrate how our neuron-specific ADNeuronNet can be leveraged to uncover new risk gene candidates and cellular pathways that help advance our understanding of molecular mechanisms underlying AD etiology.

systems biology↗

bioRxiv2026

Rapid peripheral reprogramming of myelinated afferents drives human hyperalgesia

A small burn can render a large area of skin painfully tender. This widespread hyperalgesia protects injured tissue and is typically attributed to altered spinal cord mechanisms. Whether peripheral sensory afferents directly interact to contribute to hyperalgesia remains unclear. Using single-unit microneurography, we recorded cutaneous afferents before and after inducing localized TRPV1-mediated inflammatory flare. This triggered minutes-scale reweighting of transcriptomically defined TRPV1- afferents, with divergent effects among myelinated (A{beta}-range) classes: tactile receptors showed reduced responsiveness, whereas mechano-nociceptors underwent sensitization. These changes paralleled diminished tactile sensitivity and intensified mechanical pain. Recruitment of mechanically silent branches in A{beta}-range mechano-nociceptors produced wide-field amplification of peripheral nociceptive signaling beyond the inflamed site. These findings suggest that rapid peripheral crosstalk from TRPV1+ afferents reprograms TRPV1- A{beta}-afferents and drives human hyperalgesia.

neuroscience↗

bioRxiv2026

Cross-species graph-embedding unmasks the ageing microenvironment as a key determinant of pancreatic cancer malignant cell biology and therapy response

Pancreatic ductal adenocarcinoma (PDAC) has a dismal prognosis and is characterised by an extensive pro-tumorigenic stroma. Although most PDAC cases occur in older patients, the impact of ageing on malignant-stromal interactions and therapy response remains poorly understood. Here, we established orthotopically-grafted organoid-derived PDAC models across three murine age groups to characterise changes in the PDAC stroma and malignant cells with ageing. Cross-species analyses of tumour transcriptomes using a graph-embedding approach showed that integrating mouse models of different ages better captures the diversity of human PDAC, and that aged models more faithfully recapitulate the biology of older patients with PDAC. We also demonstrated that aged PDAC models have a more inflammatory stroma than that of younger tumours, shaping the malignant cell transcriptome. Finally, graph-embedding identified IRAK4 as a candidate therapeutic vulnerability in aged, but not young, KRAS- and p53-mutant PDAC, which we validated in preclinical drug studies. These findings highlight how ageing is a critical determinant of PDAC biology and associated therapeutic vulnerabilities, which should be an important consideration when designing disease models for preclinical development of precision therapies.

cancer biology↗

bioRxiv2026

cGAS-mediated IFN-I signaling contributes to disease progression in drug-refractory epilepsy

Epilepsy is a prevalent neurological disease with a third of patients becoming non-responsive to antiepileptic drugs and developing drug-refractory epilepsy (DRE). Here we report that DRE disease progression is contributed by overactive cyclic GMP-AMP synthase (cGAS), a double-stranded DNA sensor that induces type I interferon (IFN-I) signaling. In human DRE microglia, we observe a robust IFN-I signature and the activation of upstream cGAS-STING signaling. Further, in mouse models of Dravet syndrome, a genetic form of DRE, we observe the activation of the cGAS pathway. We show that microglial cGAS can be activated by DNA released from hyperexcitable neurons. Genetic reduction and pharmacological inhibition of cGAS reduces epileptic phenotypes, glial inflammatory signatures, and neuronal transcriptomic changes, underscoring the therapeutic potential of targeting cGAS for DRE treatment.

neuroscience↗

bioRxiv2026

Deep Plasma Proteomics Coupled with Functional Genomics Reveals Drivers of Parkinson's Disease Progression and Levodopa Response

Parkinsons disease (PD) is a progressive neurodegenerative disorder lacking disease-modifying therapies, and its clinical management is limited by the absence of accessible biomarkers for tracking disease progression and treatment response. To map these complex disease trajectories, we implemented an ultra-deep plasma proteomics workflow integrating Mag-Net extracellular vesicle enrichment with Orbitrap Astral mass spectrometry to profile longitudinal samples from PD patients. This approach quantified 6,481 plasma proteins at unprecedented depth in PD studies, revealing distinct signatures directly associated with disease duration and dopaminergic therapy exposure. Candidate biomarkers were subsequently validated in an independent cohort using ELISA, demonstrating robust predictive utility in AI-driven prediction models. To uncover the mechanistic drivers underlying these systemic changes, we intersected our proteomic data with novel proteome-wide gene overexpression perturbation screens designed to identify regulators of alpha-synuclein pre-formed fibril (PFF) uptake and PFF-induced neuronal toxicity. Finally, an integrative network analysis combining three independent proteome-wide assays revealed that key pathological hubs, such as CD14, IFNG, and PLAT, are targets of currently approved pharmacological agents. Collectively, these findings provide a comprehensive, systems-level map for PD biomarker discovery and highlight druggable pathways to advance precision medicine strategies.

neuroscience↗

bioRxiv2025

The impact of downsampling on data quality, univariate measurement and multivariate pattern analysis in event-related potential research

The choice of sampling rate is a critical preprocessing step in event-related potential (ERP) research, yet its impact on different analytic approaches remains underexplored. In this study, we systematically evaluated how downsampling affects data quality measured via Standardized Measurement Error (SME), conventional univariate ERP metrics (mean amplitude, peak amplitude, peak latency, and 50% area latency), and multivariate pattern analysis (MVPA; decoding). We analyzed seven commonly studied ERP components: P3, N400, N170, N2pc, mismatch negativity, error-related negativity, and lateralized readiness potential collected from neurotypical young adults. Results showed that both amplitude- and latency-based scores were significantly affected by sampling rate changes, particularly for 50% area latency that presented increased SME (reduced data quality) at lower sampling rates. Amplitude-based measures and their corresponding effect sizes were more influenced by sampling rate variations, whereas latency-based measures were comparatively stable across different temporal resolutions. In contrast, multivariate decoding performance remained highly robust, with decoding accuracy and effect sizes showing minimal variation even at the lowest sampling rate (e.g., 64 Hz). Overall, these findings suggest that caution should be exercised when employing lower sampling rates for data quality and conventional univariate ERP analyses. Nevertheless, lower sampling rates (e.g., 64 Hz) may still be appropriate for decoding analyses, particularly in studies where precise temporal resolution is not critical. For researchers analyzing ERP data with similar components, noise levels, and participant populations as in this study, following these recommendations should yield robust statistical power.

neuroscience↗

bioRxiv2025

A high-content imaging workflow to screen for molecules that reduce cellular uptake of α-synuclein preformed fibrils

A classical pathological hallmark of many neurodegenerative diseases is the formation of protein-rich aggregates and inclusions. In Parkinsons disease (PD), -synuclein (-syn) constitutes a major protein component of pathological inclusions, termed Lewy bodies. These -syn aggregates are hypothesized to spread throughout the nervous system by cell-to-cell transmission acting as templates to amplify aggregate formation. In vitro generated -syn aggregates, commonly called preformed fibrils (PFFs), have been used to investigate a number of aspects related to -syn mediated pathology across different model systems. Here we describe a semi-automated assay to screen for small molecules that interfere with the cellular uptake and accumulation of PFFs. The assay uses dopaminergic progenitor cells (DPCs), derived from human induced pluripotent stem cells (hiPSCs). In an initial screen, we tested 1520 small molecules and identified several molecules that strongly reduce intracellular PFF load in DPCs. From these hits, candidate compounds were validated in dopaminergic neurons (DNs) to demonstrate the utility of the assay. This assay provides a robust, scalable and adaptable tool to screen for molecules that affect PFF uptake in hiPSC-derived cell models. Within the scope of this screen, it led to the identification of a set of compounds with diverse annotated targets that effectively reduce the uptake of synuclein aggregates in DPCs and DNs.

neuroscience↗

bioRxiv2025

An automated workflow for quantifying the formation of synuclein aggregates in human dopaminergic neurons

Parkinsons disease (PD) is a neurodegenerative disorder characterized by alpha-synuclein (-syn) aggregates termed Lewy bodies. To model PD pathology in vitro, preformed fibrils of -syn (PFFs), which can be taken up by cells, provide a seed that drives misfolding and aggregation of endogenous -syn, with new aggregates amplifying this process. External application of PFFs to dopaminergic neurons (DNs) increases aggregate formation, marked by -syn phosphorylation at serine 129 (pS129-syn), a pathological PD marker. Building on this, we developed an automated synuclein seeding assay to quantify new -syn aggregates in iPSC-derived DNs. Using pS129-syn as a readout, we show that PFFs elicit a time- and dose-dependent increase in pS129-syn aggregates. Our high-throughput assay further revealed that aggregate formation depends on endogenous -syn levels. Treatment with PFFs produced a greater increase in pS129-syn aggregates in iPSC DNs derived from a PD patient with a triplication in the SNCA gene, which encodes the -syn protein and which elevates total -syn levels, relative to DNs from an isogenic iPSC line from the same individual, in which the SNCA gene mutation had been corrected by CRISPR/Cas9. In contrast, no pS129-syn signal was detected in neurons in which all copies of the SNCA gene had been knocked out (KO). This high-content imaging assay for synuclein seeding offers a platform for assessing compounds and therapeutics that may impede -syn aggregate formation.

neuroscience↗

bioRxiv2025

STING_IFN_CH25H lipid axis links innate immune activation to tau pathology

Genetic risk for Alzheimers disease is strongly enriched in pathways governing microglial activation and cholesterol metabolism, yet how these processes converge to drive neurodegeneration remains unclear. Here, we identify the oxysterol 25-hydroxycholesterol (25- HC) as a pathogenic lipid downstream of the cGAS-STING-IFN pathway. In models of tau pathology, interferon signaling induces Cholesterol 25-hydroxylase (CH25H) expression in microglia. Ch25h deletion in female P301S mice suppressed tau aggregation, preserved synapses, prevented brain atrophy, and rescued memory. Mechanistically, loss of CH25H disrupted STING trafficking, attenuated IFN activation, dampened self-perpetuating microglial inflammation. Strikingly, 25-HC directly accelerated tau propagation in human iPSC derived neurons. It also disrupted lysosomal and mitochondrial lipid composition, driving cholesteryl ester accumulation and promoting apoptosis under tau-induced stress. These findings define a STING-IFN-CH25H lipid axis that bridges innate immune activation to tau pathology and toxicity, offering a tractable therapeutic pathway for inflammation-driven neurodegenerative conditions.

neuroscience↗

bioRxiv2025

Multi-stromal organoid co-cultures model pancreatic cancer and pancreatitis epithelial cell-fibroblast heterogeneity

Malignant cell-fibroblast cross-talks modulate disease progression and therapy response of pancreatic ductal adenocarcinoma (PDAC). Our knowledge of the heterogeneous nature of PDAC cancer-associated fibroblasts (CAFs) has significantly increased over the last few years. Yet, whether CAFs in PDAC differ from fibroblasts in pancreatic inflammation remains poorly understood. Chronic pancreatitis - a prolonged inflammatory state of the pancreas - is a risk factor for PDAC and is characterised by abundant fibroblasts. Thus, dissecting pancreatic fibroblast and epithelial cell reprogramming in malignancy relative to inflammation could inform new preventative, diagnostic and therapeutic strategies for PDAC. Here, we studied how pancreatic malignancy and inflammation differently shape fibroblast heterogeneity and their crosstalk with epithelial cells. We analysed human samples and mouse models of pancreatitis and PDAC and leveraged new murine pancreatitis-derived epithelial organoids to establish pancreatitis and PDAC organoid/multi-stroma co-cultures comprising pancreatic stellate cells, fibroblasts and mesothelial cells. We demonstrate that a combination of in vitro and in vivo models better captures epithelial cell and fibroblast markers of human pancreatitis and PDAC compared to mouse models alone. Finally, we identify PDAC and pancreatitis epithelial cell-specific reprogramming of stromal cells of different origin, and we infer the contribution of these distinct stromal cell types to fibroblasts in PDAC and pancreatitis in vivo. Together, our study highlights different epithelial cell-fibroblast heterogeneity in PDAC and pancreatitis, and provides new platforms for the identification of markers and epithelial-stromal interdependencies of these diseases.

cancer biology↗

bioRxiv2025

3-Phenylpropionic acid, a microbiota-derived polyphenol metabolite linked to hippuric acid, ameliorates colitis as a colon-enriched PPAR-γ agonist

Polyphenols are promising therapeutics for Crohns disease (CD), yet their bioavailability is largely dependent on the gut microbiome. The bioactive metabolites, metabolic pathways, and anti-inflammatory mechanisms underlying their effects remain poorly defined. We identified hippuric acid (HIPA) as a biomarker of polyphenol-microbiome interaction. HIPA is a meta-organismal metabolite produced through a cross-organ pathway that involves synthesis from gut microbial polyphenol metabolism via 3-phenylpropionic acid (3-PPA), followed by renal clearance. We found that CD is characterised by impaired polyphenol metabolism, resulting in deficient colonic 3-PPA and reduced serum hippuric acid relative to healthy individuals. Furthermore, we demonstrate that 3-PPA is a colon-enriched, mild PPAR-{gamma} agonist that ameliorates colitis in murine model. Systemic bioinformatic profiling of metagenomic data suggested the existence of undiscovered 3-PPA producers. Through targeted microbial incubation with polyphenols, we identified the probiotic Bifidobacterium breve and a lab isolate Escherichia coli FAH as novel 3-PPA producers. Collectively, our findings establish 3-PPA as a diet-dependent, colon-produced, mild PPAR-{gamma} agonist with the potential to minimise adverse effects associated with traditional PPAR-{gamma} agonists. This work establishes a foundation for developing microbiota-targeted, polyphenol-based therapeutic strategies for CD patients.

microbiology↗

bioRxiv2025

N-terminal acetylation reduces α-synuclein pathology in models of Parkinson's disease

The -synuclein protein, encoded by SNCA gene, is a major constituent of pathological intracellular inclusions such as Lewy bodies found in the brains of patients with Parkinsons disease and other synucleinopathies. Whereas -synuclein phosphorylation has been much studied, comparatively less work has been devoted to other post-translational modifications such as acetylation, especially given that N-terminally acetylated -synuclein is the most abundant endogenous form of the protein in the brain. In this study, using multiple in vitro and in vivo models, we sought to better understand the role of N-terminal acetylation in the pathogenesis of synucleinopathies. We found that N-terminal acetylation slowed aggregation of both -synuclein monomers and pre-formed fibrils in vitro. Uptake of acetylated -synuclein pre-formed fibrils into both immortalized cell lines and iPSC-derived dopamine neurons was also slowed compared non-acetylated fibrils. In addition, exposure to acetylated pre-formed fibrils induced less seeding of endogenous -synuclein, as measured by the accumulation of Serine129-phosphorylated -synuclein inclusions in both iPSC-derived dopamine neurons and mouse brain. Finally, mice injected with N-terminally acetylated -synuclein pre-formed fibrils survived significantly longer than mice injected with non-acetylated fibrils. Taken together, our study indicates that N-terminal acetylation reduces -synuclein aggregation, uptake into cells, seeding of endogenous -synuclein, and toxicity in vivo, suggesting that this prevalent post-translational modification represents a potent, physiologically relevant protective mechanism, which has thus far largely not been taken into consideration in most experimental paradigms of Parkinsons disease and synucleinopathies.

molecular biology↗

bioRxiv2025

Abnormally increased intrinsic neural timescales in sensory and default mode networks in cocaine use disorder

Cocaine use disorder (CUD) is associated with abnormal structural and functional brain changes. However, the neurodynamics and molecular underpinnings remain unclear. In this study, we mapped whole-brain intrinsic neural timescales (INTs), reflecting temporal neural processing, using resting-state functional magnetic resonance imaging data from 44 CUD patients and 44 healthy controls (HC). CUD showed increased INTs in visual, somatomotor, and default mode networks compared with HC. Mediation analysis linked local INTs abnormalities to altered dorsal attention network neurodynamics, associated with inhibitory control deficits. Notably, these changes were primarily correlated with alterations in gamma-aminobutyric acid type A receptors and the noradrenaline transporter. Machine learning classifiers based on INTs achieved a maximum accuracy of 75.5% in distinguishing CUD from HC, with a generalization accuracy of 65.0% on an independent dataset. This study elucidates aberrant neural mechanisms underlying CUD and highlights INTs as promising diagnostic biomarkers for clinical detection and intervention. TeaserSpatiotemporal neuroscience reveals intrinsic neural timescale disruptions underlying cocaine use disorder, offering novel diagnostic biomarkers.

neuroscience↗

bioRxiv2025

A specialized cold sensing system in the naked mole-rat

Avoiding cold or seeking warmth are universal animal needs. Homeothermic mammals maintain a constant body temperature despite fluctuating ambient temperatures. One exception is the naked mole-rat which lacks effective thermogenesis. We show that the naked mole-rat has evolved a greatly expanded cold sensing system. Compared to mice, this species has many more cold-sensitive sensory neurons and most express the cold-activated TRPM8 channel. The naked mole-rat TrpM8 gene harbors a unique upstream exon that when translated produces a TRPM8 protein with a 71 amino acid N-terminal extension. When expressed, the N-terminal extension prevents membrane targeting and abolishes TRPM8 function. Splice forms lacking the extension formed functional cold and menthol activated channels in vivo. Additionally, many naked mole-rat sensory neurons use a TRPM8-independent mechanism to detect cold. Thus, we identified molecular changes that confer both sensitivity and flexibility to cold sensing in a species that is critically dependent on following thermal cues.

neuroscience↗

bioRxiv2025

Ultrasensitive proteomics uncovers nociceptor diversity and novel pain targets

The richness of our somatosensory experience is reflected in the functional diversity of somatic sensory neurons. Single-cell sequencing (scRNA) of sensory neurons has revealed a molecular basis for such diversity1-3. However, sensory neuron diversity has yet to be captured at the level of the proteome. Here, we combined electrophysiology with deep visual proteomics (DVP)4 to quantify over 6,000 proteins from phenotypically-defined sensory neurons and identified novel proteomic markers of sensory neuron subtypes. Comparative analysis revealed both concordance and meaningful divergence between transcriptomes and proteomes. We further show that up to 3,000 proteins can be quantified from one-fourth of a single neuron, demonstrating subset-specific protein signatures. In culture, nociceptive neurons can be acutely sensitized to mechanical stimuli by nerve growth factor (NGF) which normally drives inflammatory pain in vivo5. Indeed, overnight exposure of peptidergic nociceptors to NGF and a protein kinase C (PKC) activator produced functional sensitization associated with proteome changes. Functional knockdown experiments identified the up-regulated B3GNT2 enzyme as a potential effector of nociceptor sensitization. In summary, we present a high-resolution proteomic resource linking molecular identity to function, enabling the discovery of novel mechanisms underlying somatic sensation and pain sensitization.

systems biology↗

bioRxiv2025

Integrative Multi-omics Deciphers the Assembly Rules of the Human Skin Microbiome and Highlights Lipid-Mediated Niche Partitioning

Understanding the ecological assembly mechanisms of microbial communities is critical for deciphering host-microbiota interactions, however, such processes remain poorly characterized in cutaneous ecosystems. By profiling a non-industrialized rural cohort, this study deciphers the assembly rules governing the human skin microbiome and reveals an evolutionarily conserved core microbiota structured by niche selection. Microbial -diversity peaks when C. acnes and M. osloensis reach equilibrium, supporting the Intermediate Disturbance Hypothesis. Host lipids, rather than aqueous compounds, emerged as the primary drivers of microbial niche partitioning. We further demonstrate that C. acnes and Malassezia restricta function as core metabolic engineers, expressing diverse lipases that hydrolyze triglycerides (TGs) into free fatty acids (FFAs), thereby maintaining skin surface lipid homeostasis ([~]85% TG + FFA). Notably, virtually all skin surface FFAs are microbially derived and function as "public goods", enabling cross-feeding that supports auxotrophic members and fosters mutualistic interactions. Importantly, in vitro coculture experiments confirm that C. acnes establish a survival niche for M. restricta. Our findings support a paradigm shift from a taxonomic to a functional understanding of the skin ecosystem, underscoring microbial lipid metabolism as a central orchestrator of ecological stability and a promising target for therapeutic intervention. O_FIG O_LINKSMALLFIG WIDTH=182 HEIGHT=200 SRC="FIGDIR/small/674196v1_ufig1.gif" ALT="Figure 1"> View larger version (62K): org.highwire.dtl.DTLVardef@1274fa5org.highwire.dtl.DTLVardef@1ddfa07org.highwire.dtl.DTLVardef@49f917org.highwire.dtl.DTLVardef@807a14_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGRAPHIC ABSTRACTC_FLOATNO C_FIG

microbiology↗

bioRxiv2025

Segmentation Matters: Recognizing the Cell Segmentation Challenge in Spatial Transcriptomics

Probe-based in situ hybridization spatial transcriptomics has emerged as a state-of-the-art for neuroscience research. Accurate segmentation of neurons and non-neuronal cells, a critical step for downstream analysis, remains a big challenge. Using human sensory ganglia neurons as an example, we systematically explore this problem. We evaluated multiple automatic segmentation approaches using quantitative performance metrics and downstream results. We show that careful parameter tuning is essential for achieving accurate segmentation; however, even with optimized parameters, different models still yield distinct classes of errors. To mitigate these errors, we propose a manual quality check, which can validate and refine automated segmentation results. As a future direction, integrating multi-modal imaging data with tailored neural networks may further improve segmentation accuracy. In summary, our results argue that each automated segmentation method has distinct strengths, weaknesses, and characteristic error patterns; and that a manual review is necessary.

neuroscience↗

bioRxiv2025

CREG1 restricts ALV-J replication via the mitochondrial dysfunction-driven activation of innate immunity and apoptosis

Despite the implementation of purification strategies to partially limit J subgroup avian leukosis virus (ALV-J) infection, the involvement of host factors in the underlying infection mechanism remains largely undefined. Here, we identify cellular repressor of E1A-stimulated genes 1 (CREG1) as a key regulator of mitochondrial function and a critical immune-related gene involved in ALV-J infection. The objective of this study was to explore the effects and underlying mechanisms of CREG1 in the context of ALV-J infection. Overexpression of CREG1 upregulates the expression of type I interferon (I-IFN) and certain interferon-stimulated genes (ISGs), thereby suppressing viral replication. Mechanistically, overexpression of CREG1 induces mitochondrial dysfunction, characterized by a decrease in mitochondrial membrane potential ({Delta}{psi}m), reduced adenosine triphosphate (ATP) production and respiratory chain activity, enhanced mitophagy, and increased release of mitochondrial DNA (mtDNA), which in turn triggers the activation of innate immune responses. Mitochondrial dysfunction further leads to the cytosolic release of cytochrome c and an increase in reactive oxygen species (ROS) levels, thereby triggering a robust apoptotic response. Moreover, the regulation of mitochondrial function by CREG1 depends on its interaction with the mitochondrial chaperone protein heat shock protein 1 (HSPD1), and their co-expression synergistically amplifies the antiviral response. In this study, we identify CREG1 as a potent antiviral gene and underscore the pivotal roles of mitochondria-mediated innate immunity and apoptosis during ALV-J infection. Author summaryDespite efforts to limit J subgroup avian leukosis virus (ALV-J) infection, the role of host factors in the infection process is still not fully understood. In this study, we highlight the cellular repressor of E1A-stimulated genes 1 (CREG1) as a crucial regulator of mitochondrial function and immune responses during ALV-J infection. We show that CREG1 overexpression enhances type I interferon (I-IFN) and interferon-stimulated genes (ISGs) expression, inhibiting viral replication. This is achieved through mitochondrial dysfunction, including reduced mitochondrial membrane potential, lower ATP production, and increased mitophagy and mitochondrial DNA release, which activate innate immunity. CREG1-induced mitochondrial dysfunction also triggers apoptosis by releasing cytochrome c and increasing reactive oxygen species (ROS). Furthermore, CREG1 interacts with heat shock protein 1 (HSPD1) to amplify its antiviral effects. Our findings establish CREG1 as a key antiviral gene and emphasize the importance of mitochondrial-mediated immune responses and apoptosis in ALV-J infection.

immunology↗