Biology subjects

Jiang, J.

Publications and source records attributed to Jiang, J..

At least 19 recordsLinked to original sources

bioRxiv2026

Susceptibility and cellular responses of COPD airway epithelium to cigarette smoke in stem cell-derived ALI models

Despite cigarette smoke being the chief driver of chronic obstructive pulmonary disease (COPD), numerous patients maintain their smoking habits by opting for low-yield or slim cigarettes under the misconception of diminished health risks. Differential effects of high-tar conventional versus low-tar slim cigarettes on COPD airway epithelium remain poorly defined. We established an air-liquid interface (ALI) model using patient-derived COPD airway epithelial stem cells and exposed cultures to high-tar conventional smoke (TC), low-tar slim smoke (SC), or filtered air (NC). Both regimens induced IL-1{beta} inflammation, ciliary structural changes, and oxidative stress, with TC eliciting stronger responses. Single-cell RNA sequencing resolved six epithelial populations. In ciliated cells, hdWGCNA showed NC modules associated with cilium organization and assembly, SC adaptive defense responses, and TC more extensive reprogramming of ciliary functional networks. Basal cells displayed stress-pathway activation, squamous metaplasia signatures, and developmental regression toward a stem-like state, with TC producing approximately 1.7-fold stronger regression than SC. Secretory cells exhibited identity erosion, antimicrobial program downregulation, and a shift toward keratinization and squamous-oriented programs, again more pronounced under TC. These findings indicate that high-tar conventional smoke exerts greater influence than low-tar slim smoke on COPD airway epithelium across inflammatory, structural, and lineage-specific molecular dimensions, providing high-resolution insight into cell-type-specific responses to continued smoke exposure.

cell biology↗

bioRxiv2026

Acetylation of NDUFA13 at K7 Senses OXPHOS Dysfunction to Disrupt Prohibitin Scaffolds and Drive Cristae Remodelling in Heart Failure

Mitochondrial cristae disintegration is a hallmark of heart failure, yet how oxidative phosphorylation (OXPHOS) dysfunction triggers this structural collapse remains unclear. Here we identify acetylation of the complex I subunit NDUFA13 at lysine 7 (K7) as a metabolic sensor coupling OXPHOS dysfunction to cristae remodelling. OXPHOS inhibition drove NDUFA13-K7 hyperacetylation, inducing conformational compaction and translocation to the intermembrane space. Acetylated NDUFA13 competitively engaged PHB2 through its coiled-coil domain, displacing PHB1 and disassembling prohibitin scaffolds. The acetylation-mimetic NDUFA13-K7Q induced cristae fragmentation in vitro, and cardiomyocyte-specific K7Q knock-in mice showed accelerated mortality and cristae loss under pressure overload, accompanied by impaired TCA cycle flux, defective fatty acid oxidation and selective depletion of polyunsaturated cardiolipins. These findings establish NDUFA13-K7 acetylation as a reversible switch converting bioenergetic failure into pathological cristae remodelling and metabolic reprogramming, highlighting therapeutic potential for targeted deacetylation.

cell biology↗

bioRxiv2026

Plantaricin JK from Lactiplantibacillus plantarum ZJ316 inhibits enterohemorrhagic Escherichia coli O104:H4 and alters fecal microbial communities in vitro

Enterohemorrhagic Escherichia coli (EHEC) O104:H4 is a highly virulent foodborne pathogen for which antibiotic therapy is contraindicated, severely restricting conventional treatment options; therefore, the development of novel antimicrobial strategies is essential. This study investigated the antibacterial activity and gut microbiota-modulating effects of plantaricin JK (PlnJK), a two-peptide bacteriocin from Lactiplantibacillus plantarum. Chemically synthesized PlnJK exhibited a minimum inhibitory concentration (MIC) of 32 M against E. coli O104:H4, disrupting membrane integrity as evidenced by increased extracellular conductivity, ATP depletion, and ultrastructural damage. In an in vitro fecal fermentation model, PlnJK reduced Escherichia-Shigella abundance from 36.70% to 30.60% which contains E. coli O104:H4-related taxa, enriched Clostridium (5.70% vs 2.40% in infection group), Rikenellaceae, and Lactobacillaceae, and significantly elevated short-chain fatty acid (SCFA) levels including butyrate (p < 0.01) and isovalerate (p < 0.05). To translate this into a deliverable format, CRISPR-Cas9-engineered L. plantarum strains with plnJK knockout, complementation, and overexpression were constructed. Compared with the chemically synthesized peptide, LAB-derived PlnJK--especially from the overexpression strain--showed superior efficacy in restoring microbial diversity and enhancing SCFA production, significantly increased Shannon index and reduced dominance (p < 0.01), while promoting butyrate (p < 0.01) and isovalerate (p < 0.01) production compared to the knockout control. The engineered strains further revealed a clear dose-dependent effect, as overexpression potentiated while knockout abolished the beneficial modulation. These findings demonstrate that PlnJK acts through membrane disruption and beneficially modulates gut microecology, with LAB-mediated in situ delivery offering a promising strategy for anti-EHEC live biotherapeutic development.

microbiology↗

bioRxiv2026

A qualitative and quantitative framework for the characterization of protein-RNA interactions by cross-linking mass spectrometry (XL-MS)

Protein-RNA interactions regulate nearly all levels of gene expression. Their characterization at the molecular basis is critical for understanding the function of protein-RNA complexes, both in cell homeostasis and pathology. Yet, molecular interactions underlying the formation of these complexes remain understudied. UV cross-linking mass spectrometry (XL-MS) is one of the leading approaches for in vitro identification of protein-RNA contact sites, and for providing structural information in the form of distance constraints crucial for the refinement of structural models. Our previously published CLIR-MS method enables qualitative characterization of protein-RNA interactions at residue-specific resolution but was not designed for highly accurate quantitative interpretation. Here, we present a XL-MS workflow based on irradiation with UV light that combines data-dependent acquisition (DDA) and parallel reaction monitoring (PRM) for simultaneous detection and characterization of protein-RNA interaction sites at residue-specific resolution, followed by label-free quantification of cross-linked products. We successfully apply our combined workflow to in vitro reconstituted protein-RNA complexes formed by the proteins TDP-43 and FOX1. We report on novel RNA interaction sites located in the oligomerization interface of the N-terminal domain (NTD) of TDP-43, and characterize quantitatively the process of RNA-induced monomerization of the NTD in an RNA titration experiment. We also investigate temperature-dependent differences in the efficiency of cross-link formation in the FOX1 complex. Our framework offers a unique and novel approach to comparative quantification of protein-RNA cross-links, overcoming the limitations imposed by conventional UV XL-MS methods.

biophysics↗

bioRxiv2026

Multiplexed mammalian gene control via modular splicing riboswitches

Precise, multiplexed gene regulation in mammalian cells remains constrained by a heavy reliance on heterologous protein regulators, whose associated immunogenicity and strict cargo-size limits impede clinical translation. Here, we introduce SpliceRACER (Splice-based Riboswitch Array for Combinatorial Expression Regulation), a multiplexed RNA switch platform that achieves scalable, orthogonal, and combinatorial control of gene expression in mammalian cells through alternative splicing, eliminating the need for exogenous protein co-expression. SpliceRACER independently orchestrates three orthogonal channels to regulate fluorescent reporter outputs, yielding 27 discrete combinatorial expression states, vastly exceeding the capacity of conventional binary gene switches. Furthermore, we engineered three generalizable strategies to effectively suppress the basal leakage inherent to synthetic riboswitches, thereby enabling tight OFF-state control of the potent pyroptotic effector GSDMD-NT for precise cell-death induction. This work offers broad utility in synthetic biology and therapeutic applications.

bioengineering↗

bioRxiv2026

mtDNA Activates AIM2-Mediated PANoptosis in Interstitial Macrophages to Promote Immune Checkpoint Inhibitor Pneumonitis

Immune checkpoint inhibitor pneumonitis (CIP) is a serious immune-related adverse event, yet its mechanisms remain unclear. Using a mouse model of anti-PD-1-induced CIP, we show that PD-1 blockade induces pulmonary inflammation and dysfunction, accompanied by accumulation of lung interstitial macrophages (IMs) and PANoptosis, preferentially in IMs rather than alveolar macrophages. Anti-PD-1 also causes mitochondrial stress and cytosolic mtDNA accumulation. AIM2 deficiency attenuates pulmonary inflammation, IM accumulation, and PANoptosis-associated signaling; NLRP3 or GSDMD deficiency similarly reduces inflammation and PANoptotic responses. In macrophages, TNF- plus IFN-{gamma} induces mitochondrial dysfunction and cytosolic mtDNA accumulation, and mtDNA amplifies cytokine-induced cell death and caspase-1/GSDMD activation, which is abolished by AIM2 deficiency. Thus, AIM2-mediated PANoptosis in lung IMs drives CIP, and mtDNA-associated innate immune signaling amplifies inflammatory macrophage death during CIP.

immunology↗

bioRxiv2026

Operator-valued free probability applied to recurrent neural networks with heterogeneity in local and global circuit structures

Random connectivity is a common assumption in models of recurrent neural networks, yet biological circuits combine heterogeneous synaptic connections with organization into distinct anatomical regions and functional groups. How does this global structure interact with cellular and local circuit heterogeneity to shape the dimensionality of population activity? In our work, we use operator-valued free probability to derive the covariance spectrum and effective dimension of recurrent networks with heterogeneous gains and block-structured random connectivity. This framework handles the noncommuting matrix operations through which cellular and circuit heterogeneity jointly determine population variability. We show that the linear relationship between effective dimension and firing rate uniformity persists despite global connectivity structure. At fixed total connectivity variance and with identical firing rate distributions across areas, concentrating variance within one area decreases dimension, whereas concentrating it in directed connections between areas increases dimension. Together, these results establish a mechanistic connection between circuit organization and population dimensionality, and demonstrate how operator-valued free probability unifies cellular, local, and global sources of heterogeneity in recurrent networks.

neuroscience↗

bioRxiv2026

Temporal RNA splicing dysregulation in a human multicellular model of C9orf72 ALS

Amyotrophic lateral sclerosis (ALS) is a complex and heterogenous disease, comprising of familial and sporadic forms, numerous genetic mutations, and a wide range of affected cell types and proposed dysregulated processes. While the usage of in vitro cell cultures, mouse models, and postmortem patient tissue have advanced our understanding of ALS disease mechanisms, the temporal progression of the disease and how early molecular changes evolve remain poorly understood. Here, we establish human spinal cord organoids (hSpO) derived from C9orf72 patient induced pluripotent stem cells (C9-ALS hiPSCs) as a platform to model longitudinal changes across the disease trajectory. We show that C9-ALS hSpO recapitulate hallmark C9orf72-related pathologies, increased cell death, and progressive neuronal dysfunction. We further identify RNA splicing dysregulation as an early and persistent feature of C9-ALS, with splicing signatures in hSpO closely mirroring those observed in postmortem patient tissue. Finally, prediction of RNA binding proteins (RBPs) that induce early splicing changes in C9-ALS reveals signatures consistent with a broad RBP network dysfunction. Together, this work establishes hSpO as an in vitro system for investigating early disease mechanisms of ALS in a human multicellular context and highlights dysregulated events that precede cell death.

neuroscience↗

bioRxiv2026

Pangenome-resolved polymorphic transposable elements reveal human regulatory evolution in action

Transposable elements are a major source of gene-regulatory functions. Yet this contribution is understood largely from ancient insertions already fixed in the genome: the successes, seen long after the fact. The polymorphic phase before fixation or loss remains poorly resolved because such alleles could be scored only as present or absent. We resolved segregating transposable-element alleles as sequences, epigenetic states, and transcript components using 231 haplotype-resolved human pangenomes with matched long-read multi-omics. Their origins extend beyond recent transposition; their regulatory consequences depend on methylation and transcript context and are often missed by gene-level analyses; they provide mechanisms at trait-associated loci, while selection favors element presence and absence alike. Regulatory innovation before fixation is therefore not one-way accumulation, but a mixture of gain, remodeling, and loss.

genetics↗

bioRxiv2026

An RBP-J-heparan sulfate-dependent gatekeeper directs macrophage fate between inflammatory regulation and osteoclastogenesis

Macrophages in bone possess a unique bi-potential capacity. Different from their primary roles in inflammation and immunity, these cells can differentiate into bone-resorbing osteoclasts. However, it remains unclear how macrophages determine which function to engage in bone, especially when exposed to the same stimulus, such as TNF, which can drive both pathways. Understanding these mechanisms is particularly important in inflammatory bone diseases, such as rheumatoid arthritis, where inflammation and bone resorption are hallmarks. Here, we identified an RBP-J-Hs2st1-2-O-sulfated heparan sulfate (HS)-IFN{beta} ; dependent molecular network that acts as a gatekeeper balancing macrophage fate inclinations in response to TNF. This network integrates cellular outside-in, inside-out, and relayed outside-in pathways. RBP-J deficiency in macrophages shifts TNF action from inflammatory regulation to osteoclastogenesis. Hs2st1, the exclusive biosynthetic enzyme for 2-O sulfation of HS, is a key target suppressed by RBP-J. Loss of Hs2st1 significantly reduces inflammatory osteoclastogenesis and arthritic bone erosion without affecting physiological bone mass. Elevated Hs2st1 expression and 2-O sulfation shift macrophages toward an osteoclastogenic fate, and vice versa. We further identified residues in IFN{beta} ; that are responsible for binding HS. Mutation of these sites diminished HS binding, augmented type I IFN response, and more strongly suppressed osteoclastogenesis. Our findings reveal a previously unrecognized molecular program fine-tuning macrophage fate and function, and highlight potential therapeutic strategies for inflammatory diseases with bone defects.

immunology↗

bioRxiv2026

Ageas enables time-agnostic cell fate inference from single-cell and spatial multi-omics data

Understanding cell fate decisions is fundamental to developmental biology and disease research. However, experimental lineage tracing requires genetic manipulation, which is impractical in many systems, particularly in humans. Computational approaches often rely on time-resolved measurements, which single-cell and spatial omics studies rarely provide. Here, we present Ageas, a time-agnostic transfer learning framework for cell fate inference from single-cell and spatial multi-omics data. Ageas overcomes these limitations by learning fate memory from terminal cell populations and transferring this information to progenitor or intermediate cells, enabling fate bias inference from static molecular snapshots. To enable robust generalization across molecular modalities, Ageas employs a data-adaptive ensemble strategy with automated model selection. In benchmark datasets with lineage-traced single-cell transcriptomic and epigenomic profiles, as well as spatial transcriptomics, Ageas achieves strong performance compared to existing methods. Applying Ageas to a 3D human embryo reveals a spatially organized anterior-posterior gradient of epiblast fate priming, with anterior epiblast cells biased toward ectodermal fates and posterior cells toward primitive streak-derived lineages, accompanied by regionally graded fate-associated regulatory programs. Together, these results establish Ageas as a general framework for decoding cell fate decisions from static molecular snapshots.

bioinformatics↗

bioRxiv2026

Near-infrared optoacoustic modulation of the blood-brain barrier permeability using size-tuned hyperbranched gold nanoconstructs

The blood-brain barrier (BBB) constitutes a major bottleneck for the systemic delivery of most therapeutic agents to the central nervous system. Here, we report near-infrared reversible optoacoustic modulation of the BBB permeability (NIR-ROAMBBB), leveraging endothelial tight junction targeting hyperbranched gold nanoconstructs (HBGNCs) to amplify localized optoacoustic transduction under femtosecond laser excitation. We first synthesized HBGNCs with tunable particle sizes (62-150 nm) and consistent branch morphologies via a seed-mediated growth approach, and uncovered a non-monotonic relationship between particle dimension and optoacoustic output, where the 62 nm HBGNCs generated nearly twofold stronger optoacoustic signal than gold nanorods and gold nanostars under matched excitations. Conjugation with BV11 antibodies against junctional adhesion molecule A increased HBGNC endothelial association and cerebral accumulation, enabling focal and fluence-dependent transient BBB opening (3-6 h) under 800 nm femtosecond pulsed laser excitation, as validated by in vitro trans-endothelial electrical resistance measurements, ex vivo Evans blue extravasation staining, and in vivo NIR imaging. Featuring deep tissue penetration of NIR light, robust optoacoustic conversion of HBGNCs, and negligible femtosecond laser-induced photothermal damage, this non-invasive strategy enables precise focal modulation of BBB permeability and potential drug delivery.

bioengineering↗

bioRxiv2026

Human panepigenome represents epigenomic diversity

The human pangenome captures genetic diversity beyond a single linear reference, yet conventional DNA methylome maps largely assume that the underlying CpG substrate is fixed. Here we present a first draft of the human panepigenome on DNA methylation, generated from 440 haplotype-resolved long-read methylomes spanning 26 globally distributed populations. This map jointly captures CpG presence, absence and methylation across diverse human haplotypes, revealing 12.8 million CpGs absent from GRCh38 and an average of 1.2 million variant-associated CpGs (var-CpGs) per methylome. Var-CpGs captured population-associated epigenomic variation beyond that represented by shared reference CpGs, and structural variants could create or remove entire CpG islands, frequently through mobile element insertions. Promoter var-CpG methylation was associated with transcript expression, with most associations persisting after adjustment for the underlying CpG-altering variant, whereas a subset showed evidence of mediation or variant-by-methylation interactions. CpG-altering variants were also enriched among molecular QTLs in HPRC2 and across GTEx tissues, and among clinically and pharmacogenomically annotated loci. Together, these results establish a haplotype-resolved framework in which human epigenomic diversity reflects not only variation in methylation level but also genetic variation in presence or absence of CpG substrates, providing a foundation for panepigenomic studies across tissues, populations and disease contexts.

genomics↗

bioRxiv2026

Melittin intervention induces lncRNA response, mitochondrial dysfunction, and cell proliferation in murine cervical cancer cells

Melittin exhibits antitumor activity in cervical cancer models, yet the long non-coding RNA (lncRNA) response and associated regulatory networks remain poorly understood. Here, strand-specific RNA-seq data from melittin-treated and untreated U14 murine cervical cancer cells were analyzed to characterize melittin-responsive lncRNAs and explore their potential functional associations. A total of 28,162 lncRNAs were identified, including 27,307 known and 855 novel transcripts. Differential expression analysis revealed 404 differentially expressed lncRNAs (DElncRNAs), comprising 191 upregulated and 213 downregulated lncRNAs, w most of which were predicted to localize to the cytoplasm or nucleus. Cis-target analysis identified 52 neighboring mRNAs as putative targets of 46 DElncRNAs. Functional enrichment highlighted mitochondrial electron transfer and redox-related processes, including the mitochondrial electron transfer flavoprotein complex, electron-transferring-flavoprotein dehydrogenase activity, ubiquinone binding, and quinone binding. In parallel, melittin induced mitochondrial membrane depolarization and increased intracellular reactive oxygen species accumulation in U14 cells. Co-expression analysis further identified 138 lncRNAs co-expressed with 161 mRNAs, which were enriched in chromatin remodeling, DNA replication, and DNA repair. EdU incorporation decreased with increasing melittin concentrations, indicating suppression of DNA synthesis and proliferative activity. RT-qPCR analysis confirmed the expression trends of selected DElncRNAs. Collectively, these findings demonstrate extensive remodeling of the lncRNA landscape in melittin-treated U14 cells and suggest that melittin-responsive lncRNA-mRNA networks are associated with mitochondrial redox disruption and impaired DNA synthesis. This study provides a transcriptomic framework for identifying candidate lncRNA-mRNA regulatory axes underlying the antitumor response to melittin.

cancer biology↗

bioRxiv2026

Multiview-SPIM-{micro}PIV for mapping 3C-3D blood flow within the beating zebrafish heart

Cardiac blood flow is a regulator of several important developmental and remodelling processes in the heart, including through fluid shear forces sensed by the endothelial cells lining the heart. However, optically mapping these flow fields in the complex 3D geometry of the heart is challenging even in transparent animal models such as the zebrafish. One of the main challenges is the difficulty in measuring the out-of-plane (axial) velocity component, preventing accurate mapping of the complete 3-component-3-dimension (3C-3D) blood flow velocity field; image-based techniques such as microscopic particle image velocimetry ({micro}PIV) traditionally only provide the in-plane flow components. Here we present a computational approach to achieve full time-varying 3C-3D blood flow vector mapping using a standard selective plane illumination microscope (SPIM), based on robust cardiac phase assignment, precise measurement-driven registration of sequentially acquired z-stacks, and PIV data fusion from multiple sample orientations. Our approach holds the key to understanding the complex dynamic flow fields within the developing heart, and their role in shaping cardiac development.

biophysics↗

bioRxiv2026

Concurrent AXL inhibition enhances RAS and ERK inhibitor efficacy in KRAS-mutant pancreatic and lung cancer

Resistance limits the clinical efficacy of RAS inhibitors. We applied chemical and genetic screens and identified the AXL receptor tyrosine kinase as a driver of resistance to RAS-ERK inhibition. We determined that combination treatment with the AXL inhibitor bemcentinib (AXLi) together with the RAS(ON) multi-selective tri-complex inhibitor RMC-7977 (RASi) or the ERK-selective inhibitor SCH772984 (ERKi) significantly enhanced growth suppression in human KRAS-mutant pancreatic and lung cancer models. Combined AXLi and RASi treatment of human KRAS-mutant pancreatic cell line-derived xenograft tumors synergistically suppressed ERK activation and MYC expression, and caused tumor regression. Analyses of immunocompetent mouse allograft pancreatic tumor models revealed a largely tumor cell-intrinsic response to inhibitor treatment. We identified an unexpected mechanism whereby KRAS inhibition upregulated the AXL ligand GAS6, activating AXL but inducing an AXL-dependent adaptive resistance mechanism wherein AXL antagonizes RASi efficacy. Our observations support concurrent AXL inhibition as a strategy to enhance RAS inhibitor clinical efficacy. STATEMENT OF SIGNIFICANCEOur findings identify AXL as a driver of resistance to RAS inhibitors, establishing a combination strategy to overcome resistance and enhance RAS inhibitor therapeutic efficacy in KRAS-mutant cancer by maximally inhibiting oncogenic RAS signaling.

cancer biology↗

bioRxiv2026

Association of Sleep with Anxiety in Chinese Empty-Nest Older Adults: The Mediating Roles of Physical Exercise and Social Participation

Sleep disturbances are common among older adults and are often closely associated with anxiety symptoms, which together can substantially compromise physical and mental health.A cross-sectional analysis was conducted using data from the 2017/2018 wave of the Chinese Longitudinal Healthy Longevity Survey (CLHLS). The study included 6,106 Chinese older adults (aged [≥]65 years) living in empty-nest households. Physical exercise was measured based on self-reported regular engagement in exercise, and social participation was defined as involvement in organized social activities (e.g., community or group-based events). Associations between sleep quality and duration and anxiety were estimated using multivariable logistic regression, and mediation effects of physical exercise and social participation were tested using bootstrap resampling.Among the 6,106 participants, 593 (9.7%) reported anxiety symptoms. Multivariable logistic regression analysis indicated that poor sleep quality was significantly associated with a higher risk of anxiety (OR = 3.23, 95% CI: 2.62-4.01, P < 0.001). Short sleep duration was also an independent risk factor for anxiety (OR = 1.34, 95% CI: 1.04-1.74, P = 0.025), whereas long sleep duration showed no significant association. Subgroup analyses stratified by sex, marital status, and economic status consistently revealed significant associations between sleep (both quality and duration) and anxiety, with no statistically significant interactions observed. Mediation analyses demonstrated that physical exercise significantly mediated the relationships of both sleep quality and long sleep duration with anxiety (all P < 0.01, with bootstrap confidence intervals excluding zero). Similarly, social participation significantly mediated the relationships of both short and long sleep duration with anxiety (all P < 0.01, with bootstrap confidence intervals excluding zero).In Chinese empty-nest older adults, both poor sleep quality and short sleep duration are independent risk factors for anxiety symptoms, with sleep quality exerting a stronger influence. Physical exercise partially mediated the associations between sleep quality and anxiety and between long sleep duration and anxiety, whereas social participation mediated the association between sleep duration (short and long) and anxiety. Improving sleep quality and ensuring adequate sleep duration may help reduce anxiety risk, and interventions promoting physical exercise and social participation could enhance these protective effects.

immunology↗

bioRxiv2026

Deciphering the Effect of Melittin on Murine Cervical Cancer Cells Based on Transcriptomic Investigation

Melittin, a highly active natural antimicrobial peptide derived from honeybee venom, holds immense pharmacological potential against solid tumors. However, its specific anti-tumor efficacy and transcriptomic dynamics in cervical carcinoma remain to be systematically characterized. This study evaluated the anti-cancer properties of melittin on murine U14 cervical cancer cells following by transcriptomic investigation of the underlying mechanism. Phenotypic evaluations revealed that melittin potently inhibited U14 cell viability, while wound healing assays demonstrated a profound, dose-dependent suppression of cellular migration, culminating in near-complete migratory arrest at high concentrations. Furthermore, flow cytometry quantified a dramatic, dose-dependent surge predominantly in late apoptotic cell populations. These apoptotic events were structurally corroborated by scanning electron microscopy (SEM), which revealed severe plasma membrane perforation and morphological exhaustion. Enrichment analyses indicated that the physical membrane disruption inflicted by melittin translated into a severe metabolic crisis, marked by a global suppression of ribosomal biogenesis and mitochondrial oxidative phosphorylation. Furthermore, melittin profoundly repressed the Tie2-mediated angiogenic pathway (Etv2 down-regulation) while triggering the lethal hyperactivation of the AP-1 transcriptional stress complex (Jun, Fos, Fosb). Collectively, these findings elucidate the pharmacological networks underlying melittins cytotoxicity, providing solid molecular evidence for its development as a natural therapeutic agent against cervical cancer.

cancer biology↗