Biology subjects

Liao, C.

Publications and source records attributed to Liao, C..

At least 19 recordsLinked to original sources

bioRxiv2026

Genetic determinants of Brevundimonas-mediated attenuation of plant pattern-triggered immunity

Root-associated commensal bacteria from diverse taxonomic groups can attenuate plant pattern-triggered immunity (PTI), yet the bacterial mechanisms underlying immune attenuation remain poorly understood. Brevundimonas sp. MF374 is a root-associated Alphaproteobacterium previously identified as a strong attenuator of flg22-triggered immunity. Here, to identify the genetic determinants underlying this phenotype, we conducted transposon mutagenesis and screened 4,032 MF374 mutants for defects in immune attenuation. Deletion and complementation of candidate loci identified tonB1 (BLR19_RS15045) as the strongest determinant of immune attenuation, with ppiD (BLR19_RS02880) making a weaker contribution. A panel of GUS reporter assays designed to distinguish among different mechanisms of attenuation showed that intact MF374 cells directly reduced the immune-eliciting activity of flg22 in a tonB1-dependent manner. TonB1 also contributed to attenuation of responses triggered by a distinct flg22 sequence and the endogenous plant elicitor AtPEP1. Together, these findings implicate TonB-dependent transport in attenuation of flg22-induced immunity by MF374 and reveal a previously uncharacterized mechanism of immune attenuation by a root-associated commensal.

microbiology↗

bioRxiv2026

Analysis of Rare Coding Variation Identifies New Genetic Contributors to Schizophrenia

In this study, we present the largest rare coding variant association study of schizophrenia to date, with 87,959 schizophrenia cases and 150,587 controls. We identify 16 genes at exome-wide significance: SETD1A, ZMYM2, HERC1, RB1CC1, SCAF1, XPO7, SP4, FYN, PPP3CA, CUL1, HDAC9, JARID2, ATP9A, PTK2, STAG1, and SCN2A, and an additional 24 at a 5% false discovery rate. Cases carrying ultra-rare, damaging variants, primarily protein-truncating and deleterious missense mutations, show strong enrichment in constrained genes with consistent effects across ancestries. Half of the 40 identified genes overlap with those implicated in developmental delay, autism, or bipolar disorder, underscoring shared neurodevelopmental risk. All 40 genes identified are highly expressed in excitatory and inhibitory neurons, and their expression patterns span diverse developmental stages. Functionally, the identified genes implicate chromatin regulation, protein degradation, and synaptic function. Overall, our results advance understanding of the genetic architecture of schizophrenia, provide a foundation for modeling risk genes in cellular and animal systems, and offer a framework for future work toward biologically informed patient stratification.

genetics↗

bioRxiv2026

Postnatal maturation of putamen microstructure accompanies topographic white matter connectivity and altered circuits in autism

The putamen is a major hub of the basal ganglia that emerges early in gestation. However, whether its mature organization is established before birth or emerges postnatally remains unknown. Using cross-sectional and longitudinal quantitative MRI (R1 and R2*, related to tissue density and iron, respectively) and diffusion MRI, we characterized the development of putamen's microstructure and its white matter connectivity with cortex from birth to 12 months and compared their trajectories with those in adults. Despite its prenatal emergence, the putamen undergoes substantial postnatal development. R1 increases from birth to 12 months, producing a prominent anterior-posterior gradient, whereas R2* increases primarily between age one and adulthood, producing a medial-lateral gradient. Cortico-putamen white matter connectivity is diffuse in infants but becomes topographic in adults, with anterior putamen linked to frontal cortex and posterior putamen to sensorimotor cortex. In autism spectrum disorder, this organization is largely preserved and accompanied by increased anterior putamen-prefrontal connectivity. Our findings reveal distinct spatial developmental trajectories of putamen microstructure and cortical connectivity providing a developmental framework for understanding the organization of the putamen in infancy, which has implications for assessing neurodevelopmental disorders of the basal ganglia.

neuroscience↗

bioRxiv2026

A Transferable Genomic Language Model Framework for Fungal Gene Essentiality Prediction

Predicting biological function from genomic sequence remains a major challenge in computational and systems biology. Here, we tested whether the genomic language model Evo2, which encodes context-dependent DNA sequence patterns into embeddings, enables prediction of essential genes in fungi, a phenotype central to fungal biology and antifungal target discovery. We found that model performance was constrained not by the type or complexity of the downstream classifier, but by the biological information contained in Evo2 DNA embeddings. Specifically, the information recoverable from these embeddings progressively declined for biological features further downstream of DNA sequence, revealing a bottleneck for predicting higher-order cellular phenotypes. We alleviated this bottleneck by integrating Evo2 embeddings with two sequence-informed, system-level features: ortholog-based essentiality and protein-protein interactions. The multimodal framework demonstrated consistent performance both within and across three evolutionarily divergent yeasts (Candida albicans, Saccharomyces cerevisiae, and Schizosaccharomyces pombe), and its predictions were supported by published experimental evidence when transferred to the filamentous mold Aspergillus fumigatus. These results establish our framework as a transferrable tool for predicting essential genes across fungal genomes, including species with limited or no experimentally determined essentially data.

bioinformatics↗

bioRxiv2026

A single chromosome 3p break initiates clear cell renal cell carcinoma evolution

Clear cell renal cell carcinoma (ccRCC) is initiated by chromosome 3p loss, yet chromosome losses impose a profound fitness burden on normal cells. How renal epithelial cells tolerate this deleterious aneuploidy during early tumorigenesis remains unclear. Analysis of 949 ccRCC genomes reveals two major classes of chromosome 3p alterations: simple deletions and complex rearrangements surrounding a terminal breakpoint - a pattern we term breakpoint-confined chromothripsis. We modeled both alterations in non-transformed human renal proximal tubule epithelial cells by introducing a single DNA double-strand break on chromosome 3p. Despite an initial fitness disadvantage, chromosome 3p loss drives adaptive genomic evolution that recapitulates recurrent ccRCC-associated aneuploidies, including 5q gain and 14q loss. These alterations alleviate the fitness constraints of 3p loss and promote metabolic reprogramming, clonal expansion, and malignant transformation, producing tumors with features of ccRCC. Thus, a single chromosome break initiates the evolutionary trajectory of ccRCC by creating a fitness bottleneck that selects for recurrent aneuploidies.

cancer biology↗

bioRxiv2026

Field-Correcting GRAPPA (FCG): a technique to correctspatiotemporal-varying phase errors in Echo Planar Imaging

PurposeTo develop a Field-Correcting GRAPPA (FCG) technique to correct the spatiotemporal-varying phase errors in EPI caused by eddy currents. MethodsThe fast-changing gradient in EPI causes strong eddy current effects and associated spatiotemporal-varying phase errors, producing significant image artifacts. The use of higher gradient amplitude, slew rate, and ramp sampling factor for faster imaging exacerbates this problem. In this work, FCG was developed to address this challenge by using a multi-layer perceptron (MLP) to provide a compact representation of a family of GRAPPA-like kernels that correct the spatiotemporal-varying phase errors in the data. A dedicated calibration pipeline was designed to acquire high-quality source and target data for MLP training in both slice-by-slice and simultaneous multi-slice (SMS) acquisitions. To validate FCGs assumptions and performance, a field camera was used to provide ground-truth measurement of phase patterns. The performance of FCG was further validated on phantom and in vivo experiments using demanding EPI trajectories across multiple 3T and 7T systems. ResultsField camera measurements revealed strong spatiotemporal phase variations along the kx direction that repeat along ky during EPI readouts. The experiments on high-performance systems across 3T and 7T demonstrate that FCG can provide superior correction for the artifacts induced by spatiotemporal-varying phase errors compared with existing approaches. ConclusionFCG is an effective and robust method for correcting spatiotemporal phase errors in EPI, enabling improved image quality on high-performance systems.

bioengineering↗

bioRxiv2026

Scout-based Multi-Echo NAvigator (SMENA) for high temporal resolution motion and B0 estimation and correction: applications to multi-echo GRE and EPTI

PurposeTo develop a data-driven technique, Scout-based Multi-Echo NAvigator (SMENA), for joint estimation of motion and B0 inhomogeneity ({delta}B0) at a temporal resolution of [~]200 ms with minimal additional scan time for gradient-echo acquisition. MethodsSMENA consists of two key acquisition components: SMENA-scout and SMENA-nav. SMENA-scout is a rapid 3D 4-mm multi-echo acquisition completed in less than 8 seconds, providing images with matched contrast and phase at multiple echo times. SMENA-nav captures signal variations induced by motion and{delta} B0 during the scan using compact multi-echo navigator trajectories (3.5 ms) embedded within each TR. Motion and{delta} B0 maps were jointly estimated every [~]200 ms through a model-based optimization framework relating SMENA-scout to SMENA-nav. The estimation accuracy and correction performance of SMENA were evaluated in simulations and in vivo using multi-echo GRE and GRE-EPTI acquisitions. Multiple prospective motion experiments, including large continuous movement and deep breathing, were investigated. ResultsIn both simulations and in vivo experiments, accurate motion and{delta} B0 estimation were achieved. Compared with motion-only estimation, joint estimation reduced rotation and translation errors. Joint motion and{delta} B0 correction resulted in substantial improvements in image quality, particularly at longer echo times, producing an NRMSE of 10.4% compared to 31.6% with motion-only correction. High-temporal-resolution tracking of motion and{delta} B0 enabled improved reconstruction quality in scenarios involving continuous motion and deep breathing. ConclusionSMENA enables high-temporal-resolution joint estimation of motion and{delta} B0 with minimal additional acquisition cost, providing a practical solution for motion- and{delta} B0-robust MRI.

neuroscience↗

bioRxiv2026

Gut Commensals Regulate the Intestinal Kynurenine Pathway

The kynurenine pathway (KP) of tryptophan metabolism shapes host immunity and is implicated in inflammatory, malignant, and neurodegenerative diseases. Although it is well known that inflammation influences KP activity, whether and how gut microbiota regulates KP in the intestine remains unclear. Here, we find that in allogeneic hematopoietic cell transplant patients, the kynurenine-to-tryptophan ratio tracks plasma neopterin, whereas intestinal levels do not, suggesting that local KP regulation is not explained by inflammation alone. In mice, targeted microbiota perturbation downregulates intestinal expression of indoleamine 2,3-dioxygenase 1 (IDO1), the rate-limiting enzyme of the KP, while microbiota restoration reversed this effect. Computational analyses associate Lachnospiraceae and Oscillospiraceae with colonic IDO1 in mice and inflammatory bowel disease cohorts. In vitro experiments support a mechanism whereby microbiota perturbation alters host absorption of the vitamin E isoform, {gamma}-tocotrienol, which impacts IDO1 expression. Our findings support that the gut microbiota controls colonic KP through micronutrient absorption.

microbiology↗

bioRxiv2026

Multiphasic myelination and dendritic growth modulate qMRI signals in human visual cortex.

How does myelin develop in human visual cortex? By combining immunohistochemistry with in vivo and postmortem magnetic resonance imaging of longitudinal relaxation rate (R1), which increases with myelin content, we find that myelin and R1 increase across development but follow distinct trajectories. Immunohistochemistry reveals two phases of myelination: an infant phase of limited oligodendrogenesis, with myelin restricted to deep cortical layers, followed by widespread myelination across all layers during childhood. Cortical R1 also increases across development and correlates with myelin by childhood. However, in infancy, R1 increases outpace myelin growth and instead tracks dendritic arborization, indicating that the microstructural drivers of R1 change across development. We hypothesize that deep layer myelination in infancy contributes to early visual function whereas later myelination of superficial layers enables prolonged cortical plasticity and learning of complex visual behaviors.

neuroscience↗

bioRxiv2026

A microbiota-derived bile acid overcomes antibiotic-induced hyporesponsiveness to immune checkpoint therapy by enhancing CD8+ T cell antitumor immunity

Gut microbiota are critical determinants of effective immune checkpoint therapy (ICT), yet the microbial mediators and host mechanisms that enhance antitumor immunity remain poorly understood. Here, we identify the microbiota-derived bile acid taurodeoxycholic acid (TDCA) as a metabolite associated with immune checkpoint therapy (ICT) response. TDCA administration alone is sufficient to overcome antibiotic-induced ICT hyporesponsiveness across multiple murine tumor models. Mechanistically, TDCA directly enhances CD8 T cell-mediated antitumor immunity, increasing cytotoxicity. These effects required signaling through the bile acid receptor TGR5. Together, these findings reveal TDCA as a gut microbial metabolite that restores ICT efficacy after antibiotic disruption by directly augmenting CD8 T cell anti-tumor activity. This work supports metabolite replacement as a therapeutic strategy to mitigate antibiotic-associated loss of cancer immunotherapy response. SignificanceTDCA is a microbiota-derived metabolite that restores immune checkpoint therapy efficacy after antibiotic disruption by directly enhancing CD8 T-cell-mediated anti-tumor immunity through bile acid receptor TGR5 signaling. Our findings suggest that supplementation with defined microbial metabolites can mitigate antibiotic-associated loss of immunotherapy response without requiring broader microbiome reconstitution.

cancer biology↗

bioRxiv2026

Psilocybin reshapes cortical inhibition through selective interneuron recruitment

Psychedelics show therapeutic potential for treating psychiatric disorders. While studies have emphasized the roles of cortical pyramidal cells, GABAergic neurons also express serotonin receptors and are therefore likely targets of psychedelics. In this study, we determine the effect of psilocybin on the activity dynamics of major GABAergic cell types in the mouse medial frontal cortex. Psilocybin reduces the firing of somatostatin-expressing interneurons, but increases the activity of parvalbumin-expressing interneurons. This cell type-specific response is unlikely to involve vasoactive intestinal peptide-expressing interneurons. Instead, pharmacological blockade and conditional knockout experiments demonstrate that psilocybin acts on the 5-HT1A receptor at SST interneurons, which contributes to the drugs long-term behavioral effects. Collectively, the results reveal that the classic psychedelic psilocybin alters cortical inhibition in a cell type-specific manner.

neuroscience↗

bioRxiv2026

An epigenetic mechanism of azole tolerance facilitates acquired antifungal resistance in Aspergillus fumigatus

Antibiotic tolerance paves the way for acquired resistance in bacterial pathogens. However, the mechanisms of tolerance and its evolutionary role in acquired resistance in pathogenic fungi, particularly molds, remains elusive. Here, we identified an Inhibitor of Growth domain protein (IngB) as a novel epigenetic regulator of azole tolerance in Aspergillus fumigatus. The loss of ingB promotes supra-MIC growth on agar surface despite susceptible MICs in standardized assays. Moreover, established {Delta}ingB biofilms are less susceptible to azoles in vitro and in vivo. Subsequent exposure of the tolerant strain to high azole concentrations resulted in rapid acquired resistance, most notably a frameshift mutation in a putative 20S proteasome maturation protein, UmpA, while the susceptible wildtype strain failed to acquire adaptive mutations. The data suggest that IngB-mediated tolerance provides an epistatic background for the emergence of azole resistance. Our work shows drug tolerance facilitates resistance emergence in a critical fungal pathogen. ImportanceWhile antimicrobial drug resistance causes a significant adverse effect on human health, drug tolerance can also lead to insufficient pathogen clearance, resulting in infection relapse. However, the mechanisms of antifungal drug tolerance and its evolutionary role in acquired drug resistance in pathogenic fungi, particularly the molds, remains elusive. We identified IngB as a novel regulator of azole tolerance in Aspergillus fumigatus. Importantly, loss of IngB leads to rapid azole drug resistance under azole-selective pressure. Our work identifies a novel regulator of antifungal tolerance and suggests antifungal drug tolerance can pave the way for resistance emergence in a critical fungal pathogen.

microbiology↗

bioRxiv2026

lncRNAs contribute to caste differentiation as a regulatory layer in ants

Caste differentiation in ants represents one of the major evolutionary transitions and provides a unique model for studying developmental mechanisms underlying division of labor. Genetically identical individuals follow divergent epigenetically-regulated developmental trajectories that give rise to morphologically distinct phenotypes and specialized roles. While long non-coding RNAs (lncRNAs) are emerging as key epigenetic regulators across diverse biological systems, their specific contributions to caste development in social insects remain largely unexplored. Here, we conducted comprehensive transcriptomic analyses across major developmental stages of two evolutionarily and ecologically distinct ants Monomorium pharaonis and Acromyrmex echinatior, identifying over 10,000 lncRNAs. We demonstrated that lncRNAs exhibit dynamic, caste-specific expression patterns throughout development. We also identified a subset of lncRNAs displaying canalized expression patterns, characterized by progressively increasing caste bias and decreasing within-caste variation as development proceeds. Co-expression analyses revealed that canalized lncRNAs are functionally linked to canalized protein-coding genes, which are crucial regulators for caste differentiation. These canalized lncRNAs show striking tissue-specific enrichment consistent with canalized protein-coding genes. Functional validation through RNA interference revealed that canalized lncRNAs directly regulate caste-specific traits. Furthermore, juvenile hormone treatment capable of redirecting worker lncRNA expression profiles toward gyne-like patterns, with similar expression changes and tissue specificity to JH-responsive protein-coding genes, linking lncRNA regulation to established hormonal pathways controlling caste fate. Our findings establish lncRNAs as active architects of caste differentiation in social insects, demonstrating that these rapidly-evolving regulatory molecules contribute to the evolution and maintenance of social phenotypes through tissue-specific regulation of caste-associated developmental programs.

developmental biology↗

bioRxiv2025

A high-resolution dimeric structure reveals a critical role of alpha-helix 9 in apoptotic Bax pore assembly in the mitochondrial membrane

Bax functions as a proapoptotic protein perforating the mitochondrial membrane to release mitochondrial proteins and DNA that kill the cell. Structures of full-length Bax monomer in solution and core domain dimers in solution and bound to lipid/detergent bicelles reveal how Bax proteins must be activated, unfold, refold and dimerize prior to forming oligomeric pores in the membrane. In particular, amphipathic core dimer forms part of the pore wall between the nonpolar lipid bilayer and the aqueous conduit. However, atomic resolution structures of other parts of the pore were scarce. Here, we elucidate a structure of Bax C-terminal -helix 9 (9) in lysolipid micelles using NMR. According to this high-resolution structure the 9 regions form an amphipathic helical dimer with an extended nonpolar interface and several uncharged polar residues on the surface. Structure-guided mutagenesis and functional assessment demonstrate that the nonpolar interactions are important for Bax dimerization in and perforation of the mitochondrial membrane. Surprisingly the polar residues are also important because they form bifurcated hydrogen bonds between helical turns to stabilize each helix and thereby the dimer. Molecular dynamics simulations of an oligomer constructed with wall-forming core and transmembrane 9 dimers linked by flexible 6-7-8 bridges in a mitochondrial lipid bilayer generate an atomic resolution model for a stable Bax pore capable to release cytochrome C. Thus, we made an important step toward elucidating molecular mechanisms of apoptotic Bax perforation.

biochemistry↗

bioRxiv2025

CRISPR-engineered deletion of POGZ alters transcription factor binding at promoters of genes involved in synaptic signaling

One of the seminal discoveries from genetic studies of autism spectrum disorder and related neurodevelopmental disorders (NDD) has been that loss-of-function (LoF) mutations in many genes that impact chromatin and transcriptional regulation confer substantial liability to NDD. Haploinsufficiency of the epigenetic regulator POGZ represents one of the strongest such associations; however, little is known about the direct or indirect regulatory targets of POGZ, or the mechanisms by which loss of this chromatin modifier alters early neuronal development and synaptic functions. Here, we created an allelic series of CRISPR-engineered human induced pluripotent stem cell (hiPSC) clones harboring mono- and biallelic POGZ deletions. In hiPSC-derived neural stem cells (NSC) and Neurogenin 2-induced neurons (iN), POGZ LoF altered the expression of genes associated with critical cellular processes and neuronal functions, including synaptic and intracellular signaling and extracellular matrix organization. Our multiomics profiling also showed altered footprinting of critical transcription factors (e.g., activator protein 1 complexes) that were enriched at promoters of differentially expressed genes associated with synaptic function. To further interrogate the shared molecular changes in neuronal development associated with NDD and POGZ regulation, we compared our results to deletions of the transcription factor MEF2C and the sodium channel gene SCN2A that we generated in these same isogenic iN. These analyses revealed strong enrichment of extracellular matrix and intracellular signaling disruption associated with POGZ and MEF2C deletion, whereas POGZ and SCN2A haploinsufficiency exhibited shared transcriptional effects on gene modules enriched for NDD-associated genes with opposing regulatory effects. Notably, we also observed alterations to synaptic firing rate and neurite extension with biallelic deletions, but not heterozygous lines, suggesting subtle effects in neuronal development associated with haploinsufficiency. Overall, these shared molecular consequences suggest key points of convergence that connect epigenetic regulation to neuronal function in the etiology of neurodevelopmental pathologies.

genomics↗

bioRxiv2025

Lzp Ablation Ameliorates Dyslipidemia and Suppresses Atherosclerosis by Reducing Circulating Apolipoprotein B-Containing Lipoproteins

As the principal regulator of systemic lipid homeostasis, the liver uniquely orchestrates dietary lipid assimilation, de novo lipogenesis, very-low-density lipoprotein (VLDL) assembly and secretion, and clearance of atherogenic lipoprotein remnants. This central role positions liver-specific molecular targets as critical therapeutic nodes for mitigating hyperlipidemia and halting atherosclerosis progression. The liver-specific protein Lzp (also named as OIT3), previously shown to stabilize apolipoprotein B (ApoB), the core structural component of triglyceride-rich lipoproteins, has an undefined role in vascular disease pathogenesis. Here, utilizing ApoE-/-mice, a well-established model that closely mimics human atherosclerosis, we demonstrate that Lzp deletion reduces plasma cholesterol, triglycerides, and ApoB levels under both chow and Western diets, concomitant with a marked attenuation of aortic plaque burden. Additionally, Lzp deficiency reduces hepatic and circulating ApoB levels in hyperlipidemic ApoE-/- mice, without exacerbating hepatic steatosis or injury. Mechanistically, Lzp ablation is anticipated to impair hepatic VLDL-ApoB secretion based on our previous findings, resulting in reduced circulating VLDL, intermediate density lipoprotein (IDL) and low-density lipoprotein (LDL) particles, attenuated lipid deposition, and suppressed macrophage-driven plaque inflammation. These results underscore the role of Lzp as a key regulator of systemic lipid metabolism and identify it as a potential candidate for further investigation toward therapeutic intervention of atherosclerotic cardiovascular disease. Graphic abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=128 SRC="FIGDIR/small/678175v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@118ead8org.highwire.dtl.DTLVardef@e215baorg.highwire.dtl.DTLVardef@1007212org.highwire.dtl.DTLVardef@412757_HPS_FORMAT_FIGEXP M_FIG C_FIG HightlightsO_LILzp deficiency attenuates atherosclerosis and improves plasma lipid profiles in ApoE-/- mice fed chow or Western diets. C_LIO_LILzp deficiency reduces hepatic and circulating apolipoprotein B levels in hyperlipidemic ApoE-/- mice without exacerbating hepatic steatosis or injury. C_LIO_LILzp represents a candidate therapeutic target, warranting further mechanistic and translational studies for atherosclerosis. C_LI

pathology↗

bioRxiv2025

Intracellular acidification by bacteria-derived valeric acid is a mechanism of trans-kingdom ecology against Candida parapsilosis colonization

In hematopoietic cell transplant patients, intestinal Candida parapsilosis expansion and translocation cause life-threatening candidemia, yet how commensal intestinal bacteria prevent Candida expansion remains incompletely defined. Here, we trained a machine learning model on supernatant metabolomic profiles of commensal bacteria to identify bacteria-derived inhibitors of fungal growth, with valeric and butyric acid as top hits. Experimental validation confirmed in silico predictions in three systems. First, in patient fecal samples, valeric and butyric acid levels inversely correlated with C. parapsilosis growth. Second, in vitro, valeric acid potently inhibited C. parapsilosis growth by causing intracellular acidification. Third, administration of glycerol valerate, and free or microencapsulated valeric acid blunted C. parapsilosis growth at murine intestinal sites where valeric acid could be detected. Thus, machine learning could identify a mechanistic driver of trans-kingdom ecology limiting C. parapsilosis intestinal expansion and may inform strategies to reduce patient risk of developing candidiasis during high-risk periods.

microbiology↗

bioRxiv2025

Novel Function of Arylalkalamine N-acetyltransfe- rase-1 in Modulating Host Coagulation and Blood Feeding in Aedes aegypti

During blood feeding, mosquitoes introduce substantial quantities of salivary proteins into their hosts to facilitate feeding. Arylalkylamine N-acetyltransferase-1 (aaNAT1) is a salivary protein prominently produced in the salivary glands of Aedes aegypti. Previous research has demonstrated that mosquitoes with AaaaNAT1 mutations fail to efficiently inhibit host blood coagulation; however, the precise regulatory mechanisms remain unknown. This study identified the primary metabolites and proteins interacting with AaaaNAT1 through integrated transcriptomic, proteomic, and metabolomic analyses. Combined with isothermal titration calorimetry (ITC), immunohistochemistry, ELISA, platelet function assays, and coagulation tests, we further established that AaaaNAT1 binds norepinephrine, reducing host platelet aggregation and thereby limiting activation of the host coagulation cascade and fibrinolytic system. Additionally, behavioral investigations showed that AaaaNAT1 knockdown in Aedes aegypti significantly prolonged the time required for female mosquitoes to locate a host and successfully obtain blood, while also markedly reducing blood consumption. This effect may stem from the inability of aaNAT1-depleted mosquitoes to effectively suppress host blood aggregation during feeding, as well as the downregulation of octopamine following AaaaNAT1 knockdown. Therefore, this study reveals a novel role for aaNAT1 distinct from its known functions in pigmentation and immunity, advancing our understanding of aaNAT1s role in insects.

molecular biology↗