Biology subjects

Yuan, B.

Publications and source records attributed to Yuan, B..

At least 19 recordsLinked to original sources

bioRxiv2026

Base-mismatched crRNA in CRISPR Enabled Universal One-pot Rapid Nucleic Acid Detection

CRISPR-Cas12a-based diagnostics are promising for field detection yet are often constrained by two step workflows or laborious optimization of one-pot reactions. Herein, we present a universal one-pot RPA/CRISPR-Cas12a strategy using base-mismatched crRNA in the seed region (positions 1-6). Introducing of single-nucleotide mismatches (complementary or error) into crRNA suppresses cis-cleavage activity, enabling amplification and detection in a single tube without PAM re-design or costly modified crRNA. Molecular dynamics (MD) simulations indicated that moderate potential energy (PE) variation correlates with improved screening performance, providing guidance for the selection of effective mismatches. This strategy successfully converted established two-step assays for Burkholderia pseudomallei and Brucella into one-pot formats, achieving detection of targets at 10 copies/L within 30 min with good specificity. Lyophilized pellets supported field testing on lithium-ion battery-powered devices, and experiments with simulated soil samples confirm the robustness of the approach. Together, this cost-effective, adaptable strategy can streamline RPA-CRISPR diagnostics for infectious disease surveillance.

molecular biology↗

bioRxiv2026

The chloroplast CLP chaperone-protease system controls the steady-state abundance of the singlet oxygen sensors EXCUTER 1 and 2

O_LIThe chloroplast CLP chaperone-protease is essential for chloroplast biogenesis. CLP substrate selection is aided by the N-recognin CLPS1 and CLPF adaptors. They interact with each other and the CLPC1 chaperone, but their specific functions are poorly understood. C_LIO_LIWe employed in vivo CLPC1 substrate-trapping in Arabidopsis by expressing a 35S:CLPC1-TRAP-STREPII transgene in wild-type (WT), clpf, clps1, and clpfclps1 to test the consequences of the loss of these adaptors on CLPC1-trapped proteins. Immunoblotting and protein half-life experiments were carried out for identified CLP substrates. C_LIO_LIExpression of the 35S:CLPC1-TRAP-STREPII in clps1cpf was embryo lethal. CLPF was trapped at a reduced level in clps1, supporting CLPS-CLPF interactions. Chloroplast 1O2 sensor EXECUTER1 (EX1) was trapped in WT and clps1 but not significantly in clpf. Steady-state protein accumulation of EX1 and its homolog EX2 increased 30-fold in the CLPC1-TRAP lines and clpr2-1, but not in clpf or clps1. In planta experiments showed that the half-life of EX1 is [~]3-fold longer in clpc1-1 than in WT, but EX1 half-life was unaffected in clpf. C_LIO_LIWe conclude that the CLP system plays a key role in EX1,2 homeostasis by keeping their intra-chloroplast concentrations low through continuous degradation, upstream of their 1O2 signaling function. C_LI

plant biology↗

bioRxiv2026

Systematic functional genetic analysis of cell-substrate adhesion

Cellular adhesion is critical for tissue organization and integrity, but the full complement of proteins required for proper adhesion remains unresolved. Here, we define the requirements for cell-substrate adhesion in cultured human cells using orthogonal, large-scale functional genetic approaches. Using mechanical assays to test the maintenance ("shake-off") or formation of adhesion and parallel large-scale assays of cell morphology, we identify dozens of gene targets with roles in adhesion. Our analyses reveal dynamic requirements for adhesion across timepoints and cell lines. We additionally conduct targeted downstream mechanistic analyses to resolve the molecular basis for altered adhesion. Collectively, we identify established and uncharacterized regulators of adhesion, including genes involved in mitosis, focal adhesions, actin regulation, and membrane trafficking. Unexpectedly, we find that cells that fail cytokinesis display impaired adhesion, with strongly altered actin organization and nuclear dynamics. Together, this work provides a comprehensive view of the genetic requirements for cell-substrate adhesion.

cell biology↗

bioRxiv2026

DPCGS: a computational framework for linking GWAS to single-cell transcriptomics in complex traits and diseases

Complex traits and diseases arise from the interplay between genetic variation and cellular heterogeneity, making it essential to understand how genetic risk manifests at the cellular level. However, connecting genome-wide association studies (GWAS) to specific cell populations remains challenging due to cellular complexity and the prevalence of noncoding variants. Here, we present DPCGS, a computational framework that systematically integrates GWAS summary statistics with single-cell RNA-sequencing (scRNA-seq) data to identify trait-associated cell subpopulations, genes, and regulatory programs. Unlike existing approaches that primarily evaluate pathway enrichment or cell-type-level associations, DPCGS quantifies the enrichment of genetically prioritized genes within individual cells through a statistically calibrated gene-set scoring strategy, enabling high-resolution mapping of genetic risk to cellular states. Benchmarking across simulated and diverse human single-cell datasets demonstrates that DPCGS achieves superior accuracy, sensitivity, and robustness compared with existing methods, including scDRS and scPagwas. Applying DPCGS to Alzheimers disease and asthma reveals disease-associated cellular populations and uncovers potential molecular drivers, including CD74, FOS, and AP-1 family regulatory programs, providing insights into disease-specific immune and cellular mechanisms. By bridging genetic discoveries from GWAS with functional interpretation at single-cell resolution, DPCGS establishes a generalizable framework for dissecting the cellular architecture of complex traits and diseases. This approach enables systematic discovery of disease-relevant cell subpopulations, regulatory networks, and potential therapeutic targets, offering broad applications in human genetics, single-cell biology, and precision medicine.

bioinformatics↗

bioRxiv2026

The chloroplast CLPD chaperone: consequences of under- and overexpression, interaction with the CLP protease core, and candidate substrates

Expression of the chloroplast AAA+ chaperone CLPD gene increases during senescence and drought, but its function in chloroplast proteostasis is poorly understood. This study provides a comprehensive analysis of Arabidopsis CLPD protein accumulation across development from early seedlings to senescence, and compares results to its homologs CLPC1,2, as well as CLPB3 and cpHSP90. The developmental consequences of complete loss of CLPD expression (clpd-1), as well as overexpression of functional CLPD or CLPD impaired in ATP hydrolysis (CLPD-TRAP), were determined in Arabidopsis. clpd-1 has accelerated seedling development while functional CLPD overexpression lines, but not CLPD-TRAP, have delayed development. To determine if CLPD is a bona fide CLP chaperone associating with the CLPPRT protease and to identify in vivo candidate substrates, we employed the CLPD-TRAP line during vegetative and flowering (senescent) growth stages. Affinity purification of CLPD-TRAP followed by mass spectrometry showed high enrichment of all nine subunits of the CLP protease, suggesting that CLPD plays a role in substrate delivery to the CLP protease. CLPC1,2 were also highly enriched in the CLPD-TRAP interactome, and together with prior information, suggesting hetero-oligomerization of the three chaperones. Nine chloroplast candidate substrates were identified in the CLPD-interactomes, including: FHY2 involved in riboflavin synthesis, THI1 and THIC involved in thiamin metabolism, and four proteins of unknown function. Several of these were previously identified as potential CLP substrates based on CLPC1 trapping and comparative proteomics of clp mutants. Together, this suggests that CLPD acts in substrate selection within a heteromeric CLPC-CLPD hexamer, likely with unique contributions.

plant biology↗

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

Behavioral state-dependent norepinephrine dynamics in the primary somatosensory and prefrontal cortices during tactile detection tasks

Animals must integrate sensory information, ignore behaviorally irrelevant stimuli, and respond to behaviorally relevant stimuli to find food, find mates, avoid predators, and ultimately survive. In mammals, these goal-directed behaviors require the coordinated activity of many brain regions, including sensory and prefrontal cortices and neuromodulatory brainstem nuclei like the locus coeruleus (LC), which is the brains primary source of norepinephrine (NE). NE release resulting from LC activity and arousal indexed by pupil size exert strong influences on goal-directed behavior. We explored the relationships between pupil size, cortical noradrenergic dynamics, and behavior in a tactile signal detection task. We monitored pupil dynamics and fluorescent GRABNE signals in somatosensory and medial prefrontal cortices simultaneously during task execution and found that pupil size and synchronization of GRABNE signals at baseline were strong predictors of whether animals chose to respond. Baseline and post-reward cortical GRABNE levels varied strongly with pupil-linked arousal. We also employed a generalized linear model - hidden Markov model (GLM-HMM) framework to identify distinct, stable behavioral states throughout the task that characterize task performance. We found distinct psychometric curves, task-related pupil dynamics, and cortical NE dynamics across these behavioral states. Significance StatementBehavioral state strongly shapes goal-directed behavior, which in turn depends on the coordinated activity of distributed brain regions, including the sensory and prefrontal cortices. By simultaneously measuring pupil size and cortical noradrenergic dynamics, and by identifying psychophysically distinct behavioral states during a tactile detection task, this study establishes links between pupil-linked arousal, norepinephrine signaling in somatosensory and prefrontal cortices, and trial outcomes. These findings provide new insight into how the locus coeruleus - norepinephrine system regulates perception and decision-making.

neuroscience↗

bioRxiv2025

Structural insights into the specific recognition of H2A.Z-H2B dimer by the catalytic subunit of SRCAP chromatin remodeling complex

Chromatin remodeling complexes (SRCAP-C) mediate the exchange of H2A.Z with H2A, leading to the incorporation of H2A.Z-H2B dimers into chromatin through a step-by-step manner. Here, we determined the crystal structure of the N-terminal region (531-560) of the catalytic subunit of SRCAP-C, termed SRCAP-Z domain, in complex with the engineered single-chain H2B-H2A.Z (scH2B-H2A.Z) at 2.53 [A] resolution. In the crystal structure, two molecules of scH2B-H2A.Z and one molecule of SRCAP-Z form a sandwich-like architecture. Histones form typical histone fold and SRCAP-Z domain adopts a short 310 helix. In details, Val551, Leu554, and Leu555 of the SRCAP-Z 310 helix specifically recognize the C helix of H2A.Z through hydrophobic interactions. The particular recognition has been validated in vitro by MBP-pulldown and isothermal titration calorimetry (ITC) experiments, and the MBP-pulldown assay results demonstrated that mutations in specific amino acids of SRCAP-Z attenuated its interaction with H2A.Z-H2B dimer. Finally, we demonstrated that SRCAP-Z recognizes H2A.Z-H2B dimer with a conservative structural basis and facilitates the nucleosome assembly by its chaperone activity. HighlightsFirstly, we determined the structure of the SRCAP-Z domain/scH2A.Z-H2B complex at 2.53 [A] resolution. Second, our results support a mechanism of H2A.Z-H2B/H2A-H2B exchange model in which YL1 enriches H2A.Z/H2B around nucleosomes, presenting them to SRCAP, which mediates the exchange of H2A.Z/H2B with H2A/H2B. Third, SRCAP-Z recognition of H2A.Z-H2B dimer is structurally conserved. Fourth, a distinct sandwich-like architecture: SRCAP-Z adopts a 310 helix that inserts precisely between H2A.Z and H2B, forming a stable, unexpected structural module.

biochemistry↗

bioRxiv2025

Mitochondrial control of fuel switching via carnitine biosynthesis

Environmental adaptation often involves a shift in energy utilization toward mitochondrial fatty acid oxidation, which requires carnitine. Besides dietary sources of animal origin, carnitine biosynthesis from trimethyllysine (TML) is essential, particularly for those who consume plant-based diets; however, its molecular regulation and physiological role remain elusive. Here, we identify SLC25A45 as a mitochondrial TML carrier that controls carnitine biosynthesis and fuel switching. SLC25A45 deficiency decreased the carnitine pool and impaired mitochondrial fatty acid oxidation, shifting reliance to carbohydrate metabolism. Slc25a45-deficient mice were cold-intolerant and resistant to lipid mobilization by GLP1 receptor agonist (GLP-1RA), rendering them resistant to adipose tissue loss. Our study suggests that mitochondria serve as a regulatory checkpoint in fuel switching, with implications for metabolic adaptation and the efficacy of GLP-1RA-based anti-obesity therapy.

biochemistry↗

bioRxiv2025

COTree: A Statistical Framework for Deciphering Cell-Resolved Multi-Omics Trajectories

Recent advances in whole-cell modeling enable the computational tracking of the temporal evolution of thousands of molecular species across genomic, transcriptomic, proteomic, and metabolomic layers. These models provide a complementary perspective for studying cellular dynamics, offering continuous, system-wide observations that are difficult to obtain from experimental technologies, which are often destructive and yield only static measurements from limited modalities. While whole-cell models generate multi-omic simulation trajectories with high temporal resolution, analyzing and interpreting such complex data remains a major challenge that limits their potential to elucidate cellular dynamics. To address this challenge, we propose COTree, a statistical framework that learns integrated multi-omic representations and constructs a trajectory principal tree to summarize cellular progression patterns. COTree enables a broad range of downstream analyses, including cell classification, fate prediction, developmental time detection, and driver species identification, that provide new insights into how cells develop and differentiate. To demonstrate its practical utility, we apply COTree to a multi-omic trajectory dataset generated from the whole-cell model of JCVI-Syn3A, revealing cell types, characterizing long-term cellular dynamics, and identifying key driver species associated with cell death and replication.

bioinformatics↗

bioRxiv2025

Stochastic Modeling and Transient Analysis of Epidemic Extinction Dynamics

Understanding extinction probabilities in branching processes is pivotal for epidemiology and population dynamics. Traditional models often assume a fixed generation time, resulting in extinction probabilities determined solely by offspring distributions and remaining unchanged over time. By contrast, our study incorporates the generation time into the analysis, considering how the timing of each generation influences extinction dynamics. By focusing on the finite-time risk of extinction, our approach reveals that shorter generation times can lead to a temporary increase in the risk of population or epidemic die-out. We support our findings with precise fixed-point analyses and numerical integration techniques based on real data from various infectious diseases. Although the long-term probability of extinction does not change, the transient dynamics show that a faster-paced transmission process may elevate early extinction risk. The study highlights the crucial role of transmission timing in epidemic modeling and indicates that accounting for generation time can provide new perspectives for developing effective public health strategies and outbreak control measures.

systems biology↗

bioRxiv2025

Dynamic Meta-Networking Identifies Distinct Network Correlates of Positive and Negative Formal Thought Disorder in Schizophrenia

Formal thought disorder (FTD) is a core symptom of schizophrenia, yet the neural network mechanisms underlying this phenotype remain poorly understood. In this study, we applied a dynamic meta-networking framework, which captures temporally recurring functional network states, to investigate alterations in the language and executive control networks and their associations with positive and negative FTD. Resting-state fMRI data were collected from three independent cohorts: a discovery cohort comprising 150 first-episode, drug-naive patients with schizophrenia and 175 healthy controls (HCs); a replication cohort including 183 first-episode, drug-naive patients and 109 HCs; and a third cohort consisting of 71 patients who had received two weeks of antipsychotic treatment and 71 HCs. Meta-networking analysis identified four distinct resting-state meta-states within both the language and executive control networks. Connectivity-behavior correlation analyses and machine-learning regression models revealed that positive FTD was associated with aberrant connectivity across specific meta-states in both networks. In contrast, negative FTD was linked exclusively to dysfunction within two meta-states of the executive control network. Notably, these polarity-specific, multi-state connectivity disruptions normalized following short-term antipsychotic treatment, highlighting their potential as clinically relevant neuroimaging biomarkers.

neuroscience↗

bioRxiv2025

Metabolite Profiling of Epimedium sagittatum Bee Pollen: Identified Bioactive Flavonoids with Multi-Disease Functional Food Potential

Epimedium sagittatum bee pollen (EBP) is a bee pollen product of a medicinal plant, but its chemical characteristics remain unclear. In this study, EBP was systematically characterized by biochemical analysis and UPLC-MS/MS, using Brassica rapa bee pollen (BBP), Camellia sinensis bee pollen (CBP), and Epimedium leaves for compari-son. A total of 1,073 secondary metabolites were identified in EBP, mainly flavonoids (330, 30.8%) and phenolic acids (165, 15.3%). EBP showed the highest total flavonoid content among the three bee pollen types (4.75 mg/g) and was clearly separated from BBP and CBP in multivariate analysis. EBP contained 19 unique metabolites, fewer than CBP (271), but these included several high-content flavonoid compounds, such as cacticin, brassicin, and tricetin-4'-methyl ether-3'-{beta}-D-glucoside. Differential metabolite analysis identified 449 and 1,085 metabolites that differed between EBP and BBP, re-spectively, with flavonoid compounds forming the main differential class. Among the shared differential metabolites, 45 flavonoids were consistently higher in EBP, in-cluding kaempferol, tamarixetin, and sinensetin. Network pharmacology screening further suggested that flavonoid metabolites, especially kaempferol, tamarixetin, and sinensetin, deserve particular attention in future studies of EBP. Compared with Epimedium leaves, characteristic leaf flavonoids such as icariin and epimedins A-C were present at very low levels or were not detected in EBP. Overall, EBP is charac-terized by a distinct, relatively concentrated flavonoid chemical profile, distinct from both common commercial bee pollens and Epimedium medicinal tissues. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=160 SRC="FIGDIR/small/679240v2_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@1587d35org.highwire.dtl.DTLVardef@8461d6org.highwire.dtl.DTLVardef@1336149org.highwire.dtl.DTLVardef@381ae8_HPS_FORMAT_FIGEXP M_FIG C_FIG

plant biology↗

bioRxiv2025

Beyond the Anterior Temporal Lobe: Domain-Related Degeneration of Cortical Language Network Dynamics in Semantic Dementia

Semantic dementia (SD) is a neurodegenerative disorder marked by a progressive decline in semantic memory, mainly due to focal atrophy in the anterior temporal lobe (ATL). As the disease advances, atrophy spreads to the perisylvian regions, accompanied by non-semantic language impairments. Despite these clinical findings, the network mechanisms behind cross-domain linguistic deficits remain poorly understood. In this study, we used our recently developed meta-networking framework of cortical language network dynamics to systematically examine domain-specific network disruptions in SD. Using resting-state functional MRI (fMRI) and comprehensive neuropsychological tests across several language domains, we analyzed data from SD patients at two timepoints: baseline (n = 42) and a 2-year follow-up (n = 24). Our findings showed that the framework successfully identified domain-specific language network degeneration. Beyond the ATL, progressive atrophy disrupted the dynamic separation of language networks involved in semantic processing, phonological processing, and speech production. These disruptions were characterized by state-specific hypo- and hyper-connectivity patterns that related to distinct language impairments. At follow-up, atrophy extended to posterior temporal and prefrontal regions, worsening network function. Importantly, the patterns of language network disruption predicted individual language deficits, providing a mechanistic link between structural degeneration, functional network changes, and clinical symptoms.

neuroscience↗

bioRxiv2025

The Disruption and Normalization of Cortical Language Network Dynamics After Post-Stroke White Matter Disconnection

The human language network undergoes reorganization across different spatiotemporal scales, limited by its fixed structural neurobiological foundations. Although stroke often damages white matter, its effect on temporary language network reorganization is not fully understood. This study examined longitudinal behavioral and resting-state fMRI data from post-stroke patients with exclusive subcortical lesions at three time points: two weeks (n = 38), three months (n = 29), and one year (n = 27). Patients showed mild to moderate language impairments during the acute phase, which improved within three months. The extent of disconnection in several left-hemispheric white matter tracts was negatively associated with language deficits. Healthy controls (HC, n = 25) exhibited domain-segregation dynamics in the cortical language network, limited by the underlying white matter pathways. In patients, severe, state- and track-specific network disruptions were observed in the acute phase, characterized by hypo- and hyper-connectivity and abnormal topological features. As language recovery progressed, connectivity patterns began to return to normal, resembling those of HC, and the domain-segregation dynamics reappeared. These results deepen our understanding of structure-function coupling and indicate significant post-lesional plasticity within the cortical language network.

neuroscience↗

bioRxiv2025

Polyamines buffer labile iron to suppress ferroptosis

Polyamines are essential and evolutionarily conserved metabolites present at millimolar concentrations in mammalian cells. Cells tightly regulate polyamine homeostasis through complex feedback mechanisms, yet the precise role necessitating this regulation remains unclear. Here, we show that polyamines contribute to endogenous buffering of redox-active iron, providing a molecular link between polyamine metabolism and ferroptosis. Using a genome-wide CRISPR screen, we identified a synthetic lethal dependency between polyamine depletion and the key ferroptosis suppressor, GPX4. Mechanistically, we show that polyamine deficiency triggers a redistribution of cellular iron, increasing the labile iron pool and upregulating ferritin. To directly visualize this iron buffering in living cells, we developed a genetically encoded fluorescent reporter for redox-active iron. Live-cell analysis revealed a striking inverse correlation between intracellular polyamine levels and redox-active iron at single-cell resolution. These findings reposition polyamines as key regulators of iron homeostasis, with implications for ferroptosis-linked disease states and cellular redox balance.

cell biology↗

bioRxiv2025

Molecular insights into the capsular polysaccharide transporter Wza-Wzc complex

Capsular polysaccharides (CPS), which form the protective outer capsule surrounding many Gram-negative bacterial pathogens, are critical virulence determinants. Their biosynthesis is primarily carried out via the conserved Wzx/Wzy-dependent pathway. In Escherichia coli, Group 1 CPS transport through the bacterial envelope is thought to be mediated by the Wza-Wzc complex. In this study, we present the first structural characterization of the complete Wza-Wzc complex from E. coli K12, determined using single-particle cryogenic electron microscopy. The structure revealed an elongated, continuous channel spanning the entire envelope, which is crucial for efficient CPS secretion, as supported by mutagenesis studies. Multiple structural snapshots of the ADP-bound Wza-Wzc complex captured intermediate conformations of the double membrane assembly, highlighting its remarkable intrinsic dynamics. In-depth analysis of the isolated Wza translocon and Wzc co-polymerase, revealed new mechanistic details of both complex formation and CPS transport. Importantly, we identified the jellyroll domain of Wzc as a previously unrecognized CPS-binding module, likely guiding CPS repeat units into a proposed Wzc-Wzy polymerization platform. Collectively, these findings provide new structural and functional insights into CPS synthesis and transport, advancing our understanding of bacterial capsule formation and virulence mechanisms.

microbiology↗

bioRxiv2025

Cholesterol-Responsive Interaction of NFE2L1-INSIG1 Controls VLDL Secretion and MASH Pathogenesis

Cholesterol overload drives metabolic dysfunction-associated steatohepatitis (MASH). While the liver maintains homeostasis by exporting cholesterol via very low-density lipoprotein (VLDL) secretion, this process is paradoxically suppressed under cholesterol excess by insulin-induced gene 1 (INSIG1), which inhibits sterol regulatory element-binding protein 1 (SREBP1) activity. How VLDL secretion persists during cholesterol overload to prevent lipotoxicity remains unresolved. We identify a cholesterol-responsive interaction between nuclear factor erythroid 2 related factor-1 (NFE2L1) and INSIG1 that sustains cholesterol balance. Hepatic NFE2L1 deficiency elevates INSIG1 levels, suppressing SREBP1 activation and impairing VLDL secretion, leading to cholesterol accumulation and liver injury. Mechanistically, NFE2L1 binds to INSIG1 via its N-terminal homology box 2 (NHB2) domain, with cholesterol enhancing this interaction to drive INSIG1 degradation and SREBP1 activation. In NFE2L1-deficient mice, wild-type NFE2L1 restores SREBP1 activity and VLDL secretion, while the NHB2-deleted mutant ({Delta}NHB2) fails. Lipidomics reveal that NFE2L1 deficiency reduces serum triglyceride composition, restored exclusively by wild-type NFE2L1. In a murine MASH model, NFE2L1 overexpression activates SREBP1/2, enhances cholesterol secretion, and alleviates liver injury, inflammation and fibrosis, without elevating atherogenic lipoproteins due to compensatory LDL receptor upregulation. Our findings resolve the paradox of cholesterol-driven VLDL secretion and establish the NFE2L1-INSIG1 axis as a therapeutic target for metabolic diseases.

physiology↗