Biology subjects

Chen, D.

Publications and source records attributed to Chen, D..

At least 19 recordsLinked to original sources

bioRxiv2026

Mental replay rescales the subjective duration of past events via alpha and theta oscillations

Human memory recall is flexible in time, allowing past events to be mentally replayed at variable speeds. However, it remains unclear whether temporally rescaled replay distorts the perceived duration of remembered events, and whether such distortion is tied to changes in the retrieved content. Here we instructed participants to mentally replay previously encoded naturalistic videos at compressed, expanded, or self-paced (free) speeds while recording scalp electroencephalography (EEG). Replaying an event at an expanded pace lengthened participants' subsequent judgments of its original duration relative to free and compressed replay, and the longer an event had been replayed, the longer it was judged to have lasted. Crucially, recalled memory content was unaffected, with gist accuracy and recalled detail comparable across speed conditions. During replay, alpha and theta power were reduced under compression, and both interacted with the speed instruction in predicting reported duration. Specifically, alpha power exhibited a stronger positive relationship with duration judgments under compressed replay than under free or expanded replay. Conversely, the relationship between theta power and reported duration was significantly more positive under expanded replay than under free or compressed replay. Neural pattern similarity between instructed and later spontaneous replay varied with replay speed, yet this reinstatement neither predicted reported duration nor mediated the oscillatory effects. These findings suggest that remembered duration is not read out from the amount of content retrieved but reconstructed at retrieval through oscillatory dynamics, decoupling the subjective scaling of time from the content of memory.

neuroscience↗

bioRxiv2026

Glymphatic System Is Enhanced in Anti-NMDAR Encephalitis and Facilitates Antibody Influx and Clearance in the Brain

ABSTRACT Background: Anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis is an antibody-mediated neuroinflammatory disease. How pathogenic autoantibodies enter and are cleared from the brain parenchyma remains poorly understood. The glymphatic system (GS) mediates cerebrospinal fluid (CSF)-interstitial fluid (ISF) exchange and waste removal, yet its role in anti-NMDAR encephalitis is undefined. Methods: We used polypeptides GluN1356-385 induced active-immunization anti-NMDAR encephalitis mouse model. Glymphatic influx and efflux were evaluated via fluorescent tracer delivery combined with real-time imaging and ex-vivo quantification. Perivascular aquaporin-4 (AQP4) polarization, brain IgG deposition, NMDAR levels and blood-brain barrier (BBB) integrity were analyzed. GS function was pharmacologically modulated with AER-271 (an AQP4 inhibitor) or dexmedetomidine (Dex), and deep cervical lymphatic vessels (dCLV) were ligated in a passive-transfer model (PTM). Behavioral performance was characterized by multiple neurobehavioral tests. Results: Both glymphatic influx and efflux were enhanced in anti-NMDAR encephalitis mice, accompanied by upregulated antibody entry into and clearance from the brain. However, the cerebral IgG deposition was increased. These changes were associated with increased perivascular AQP4 polarization, with no alterations in overall AQP4 abundance or BBB tight-junction integrity. AQP4 inhibition with AER-271 attenuated glymphatic influx and efflux, reduced hippocampus IgG deposition, and partially ameliorated behavioral deficits without restoring hippocampus NMDAR levels. Conversely, dCLV ligation in the PTM mice increased brain retention of patient-derived IgG and exacerbated anxiety-like behavior. Dex enhanced antibody clearance and reduced hippocampus IgG deposition but did not improve behavioral outcomes. Conclusion: The GS is dynamically upregulated in anti-NMDAR encephalitis, driven by increased perivascular AQP4 polarization. The enhanced glymphatic influx facilitates antibody entry into the brain parenchyma and exacerbates neuronal injury, whereas enhanced efflux provides a compensatory route for antibody clearance. Rather than broadly enhancing glymphatic activity, precise tuning of this pathway may present a promising non-immunological therapeutic strategy for autoimmune encephalitis. Keywords: anti-NMDAR encephalitis; antibody influx and clearance; glymphatic system; AQP4 polarization; AER-271; dexmedetomidine

neuroscience↗

bioRxiv2026

Sleep reactivation shapes memories of event and its temporal structure

Episodic memory provides a temporal scaffold that organizes past experiences according to their sequential occurrence. While memories are reactivated and consolidated during sleep, it remains unknown how exogenous reactivation reshapes memories of complex events, particularly the temporal structure that binds individual elements across time. In this preregistered study, 35 participants encoded neutral and aversive narratives composed of image sequences, with auditory context shifts defining event boundaries. During post-encoding slow-wave sleep, closed-loop targeted memory reactivation (TMR) delivered boundary-associated auditory cues time-locked to the up-state of slow oscillations. Reactivation led to faster, more confident, and more liberal recognition responses for cued than uncued memories. However, reactivation exerted a time-dependent effect on temporal-order memory, impairing it immediately after sleep but attenuating forgetting one week later. This behavioral dissociation was paralleled at the neural level: stronger cue-elicited theta activity predicted superior recognition of neutral, but not aversive event content, whereas greater theta responses were associated with poorer temporal-order memory for the same events. Together, these findings suggest that sleep reactivation does not uniformly strengthen episodic memories. Rather, it selectively prioritizes access to event content while reorganizing temporal relations among event elements, revealing distinct, time-dependent consequences of sleep reactivation for different components of complex experiences.

neuroscience↗

bioRxiv2026

Self-extending DNA recording with stochastic and programmed writing

Recording cellular histories requires new information to accumulate without erasing earlier records. Here we introduce Infinite Printer, a CRISPR recording architecture that couples self-extension to regeneration of a writable terminal context. A self-targeting prime-editing guide RNA (stpegRNA) directs repeat extension, while base editing generates diversity in situ without a separate insertion template for each character state. Earlier sequence remains upstream of the renewed terminus, linking substrate growth with record retention. Auxiliary pegRNAs specify payloads within a write-renew-write architecture. After three rounds of payload input, 4,436 distinct complete CCA-based records contained identities from all three pools. Longer AC/AT records retained shared upstream marks, and full-length sequence analysis resolved additional endpoint distinctions. Joint single-cell readout recovered 1,549 distinct records and 7,669 combinations across 12,302 transcriptome-linked cell labels from a single stpegRNA design. Spatial-transcriptomic readout recovered spacer records at 11,330 capture spots, linking record content to tissue coordinates and gene-expression profiles. Infinite Printer unites substrate renewal, sequence diversification and programmed writing with cell- and tissue-linked readout, providing a foundation for future lineage recording across large cell populations during mammalian development.

synthetic biology↗

bioRxiv2026

Spontaneous Hemodynamic Oscillations during Sleep Coordinate Directional CSF Flow in the Subarachnoid Space and Ventricles

During sleep, cerebrospinal fluid (CSF) flow is enhanced and thought to facilitate clearance of metabolic waste from the brain. Human imaging has shown that sleep is associated with large waves of bulk CSF flow in the fourth ventricle; however, the CSF flow dynamics around brain tissue are unknown. Here, we investigated how CSF flow is organized across the subarachnoid space, a key site both for fluid exchange across cortical surface and for transport out of the cranium. We used an MRI technology at 7 Tesla that simultaneously measures cortical hemodynamics and slow subarachnoid CSF flow and applied this during sleep. We found that spontaneous infraslow hemodynamic oscillations were coupled to spatially structured, directional CSF flow throughout both ventricular and subarachnoid compartments. Hemodynamic oscillations and fourth-ventricle bulk flow were both associated with brain-wide CSF flow at different phases, indicating that they reflect a single coordinated CSF movement cycle. Compared to cardiac pulsations, a canonical driver of CSF movement, hemodynamic-driven CSF flow showed a distinct directional pattern and was smaller in instantaneous amplitude but produced substantially greater cumulative displacement and thus greater potential for transport within an oscillation period. Together, these findings show that hemodynamic oscillations during sleep are coupled to temporally coordinated and directional CSF flow throughout the ventricles and subarachnoid space, suggesting that sleep-dependent neuronal activity drives fluid movement across the human brain in a distinct spatial pattern.

neuroscience↗

bioRxiv2026

Robust V(D)J Recombination in the Absence of SA1/2-mediated Chromatin Loops

The cohesin ring complex organizes and mediates functional activity of the genome through chromatin loop extrusion1,2. Cohesin comprises a core SMC1-SMC3-RAD21 ring associated with multiple regulatory proteins, including SA1 and SA2 paralogs that bind mutually-exclusively to a RAD21 site3,4. Functional studies of cohesin complexes in vivo often employ inactivation of particular components2,3. However, inactivation of SA1 or SA2 in mice is limited by mouse embryonic lethality5-7 and, for dual SA1/2-inactivation by cycling cell lethality8-10. For these reasons, it remains unknown whether the requirement for SA1/2 in loop extrusion in vitro11,12 applies in vivo1,2, and whether these factors have overlapping or distinct functions in regulating this process3. Here, we employ G0/G1-arrested v-Abl pro-B lines, which are viable in the absence of cohesin-complex factors13,14, to dissect SA1 and SA2 functions in V(D)J recombination, a strictly G0/G1-specific process that harnesses loop extrusion to effect distinct Igh and Ig{kappa} locus recombination mechanisms15,16. Robust long-range Igh or Ig{kappa} V(D)J recombination continued unabated in the absence of SA1 or SA2. However, inactivation of both abrogated most positioned chromatin loops genome-wide and also abolished long-range Igh V(D)J recombination with a phenotype similar to RAD21 (cohesin) inactivation13. Yet, robust Ig{kappa} V(D)J recombination continued upon dual SA1/2-inactivation, which along with Hi-C findings, demonstrates ongoing loop-extrusion in the absence of SA1/2. Correspondingly, replacement of weak Igh V(D)J recombination signal sequences (RSSs) that target V(D)J recombination activity, with strong Ig{kappa} counterparts15 restored long-range Igh V(D)J recombination in SA1/2 inactivated cells. We conclude that SA1/2 together maintain loop extrusion impediments and genome-wide architecture in G0/G1-arrested cells; and, in their absence, loop extrusion proceeds unimpeded. Remarkably, strong RSSs allow Ig{kappa} to function normally in the absence of internal positioned loop structure across the 3.1-mega-base Ig{kappa} locus. We discuss implications of our findings for long-range gene regulation in G0/G1-arrested cells more broadly.

immunology↗

bioRxiv2026

Repeated malaria exposure reprograms Vδ2+ γδT cell chromatin accessibility and immune function toward a reversible hyporesponsive state

V{delta}2+ {gamma}{delta} T cells contribute to antimalarial immunity via pro-inflammatory and cytotoxic mechanisms. Among children who have experienced repeated episodes of malaria, V{delta}2+ T cells develop reduced pro-inflammatory function through mechanisms that remain unknown. We hypothesized that recurrent malaria drives coordinated epigenetic and transcriptional remodeling of these semi-innate cells. Using PBMC from Ugandan children in cohorts spanning high and low transmission (10 children) and longitudinally across a marked decline in transmission (20 children, 3 timepoints), we profiled the epigenome and transcriptome of V{delta}2+ T cells by paired bulk ATAC-seq and RNA-seq and by CITE-seq. Higher malaria exposure was associated with reduced V{delta}2+ T cell TNF and IFN{gamma} production and with thousands of differentially accessible chromatin regions. Immunoregulatory genes including LYN, PLCG2, and HAVCR2 (Tim-3) showed increased chromatin accessibility and/or expression at high exposure, whereas pro-inflammatory gene (IFNG, GZMK, IL7R) and AP-1 (JUN/FOS) motif accessibility were increased at low exposure; linkage of regulatory elements with target genes corroborated these associations. Single-cell transcriptional analysis identified a cytotoxic CD8+CD16+CD57+ V{delta}2+ T cell subset. In our in vitro model, repeated stimulation with malaria-infected red blood cells or the phosphoantigen HMBPP was similarly associated with reduced V{delta}2+ T cell degranulation and cytokine production. Diminished responsiveness was predominantly TCR-dependent, accompanied by heightened p38 signaling, and was partially reversed by rest induced by the tyrosine kinase inhibitor dasatinib. Together these data reveal how repeated malaria may reshape cellular immunity by reprogramming V{delta}2+ T cells toward a reversible, hyporesponsive state and highlight targets for therapeutically restoring their function.

immunology↗

bioRxiv2026

Genomic SEM reveals shared genetic architecture and expands locus discovery for locomotor activity in heterogeneous stock rats

Locomotor response to a novel environment is a heritable, addiction-relevant behavioral phenotype in rodents. We characterized its genetic basis in 9,341 male and female heterogeneous stock (HS) rats from eight studies. Locomotor activity was heritable across studies, while pairwise genetic correlations were positive but variable, indicating shared yet nonidentical genetic influences. Given nonuniform genetic architecture, we compared three complementary approaches to locus discovery that analyzed SNP effects 1) on a pooled locomotor phenotype (MEGA), 2) across studies under a fixed-effect model (METAL), or 3) on a shared genetic factor (Genomic SEM). To ensure comparability, all three locus-discovery approaches were applied to a common subset of seven studies. Together, these approaches identified 38 loci, with both convergence and method-specific discovery across analyses. Genomic SEM identified 31 loci, compared to 8 for MEGA and 13 for METAL. To interpret the loci, we integrated brain molecular QTLs, predicted coding and splice consequences, and open-field locomotor phenotypes from knockouts of orthologous mouse genes. Our results highlighted established neurobehavioral candidates spanning locomotor and reward-related biology (Negr1 and Lmo4), serotonergic signaling (Htr1b), and GABAergic signaling (Gabrg2), as well as relatively novel genes including Tcerg1l, Pcdh10, and Tmem132e. Our results illustrate how Genomic SEM can integrate genetically correlated yet heterogeneous behavioral measures and enable discovery of loci underlying their shared genetic architecture, highlighting its utility for complex-trait studies in rodents.

genetics↗

bioRxiv2026

Joint protein, mRNA and DNA sequence design and optimization with nucleotide-level Potts models

Protein design typically generates amino acid sequences first and then translates them into DNA post-hoc limiting control over the nucleotide sequence via codon choice at design time. We introduce NuCaliby, a structure-conditioned model that jointly designs amino acid, mRNA, and DNA sequences of a protein at nucleotide resolution. NuCaliby predicts a Potts model over a nucleotide graph, trained using amino acid supervision by marginalizing over synonymous codons. It preserves residue-level designability on par with the original Caliby model. Its energy-based formulation supports composable inference-time guidance from sequence-level objectives through proposal construction and candidate acceptance, allowing the incorporation of biological proxies into the coding nucleotide sequence. Under organism-specific tRNA adaptation guidance, NuCaliby improves adaptation to host tRNA pools beyond synonymous sequence space, trading-off self-consistency. NuCaliby also embeds specified RNA motif sequences directly into coding sequences, avoiding the combinatorial growth required for the same task at the amino-acid level. In wet lab experiments, NuCaliby produces expressible and soluble proteins on par with state of the art methods, while enabling more complex nucleotide-level control and optimization. Together, these results extend protein design beyond the amino acid level, enabling joint optimization across the central dogma of molecular biology. The code and model weights are available here: https://github.com/blazejba/NuCaliby

biophysics↗

bioRxiv2026

Motor learning refines premotor cortical constraint during skilled forelimb behavior

Motor learning transforms variable actions into reliable skilled behavior, but how premotor cortex supports this transformation remains unclear. We longitudinally recorded bilateral local field potentials and single-unit activity from the rat rostral forelimb area while animals acquired a single-pellet retrieval task. Here, we show that improved skill was accompanied by reduced, rather than increased, task-related premotor recruitment. Despite this attenuation, firing remained dominated by suppression around pellet retrieval, and stronger pre-retrieval suppression was associated with greater odds of successful performance. Population neural state analysis further showed that successful actions were most strongly coupled to sustained pre-retrieval engagement early in learning, whereas later success occurred with less population-level engagement. These findings reveal premotor cortical constraint as a population-level process that is progressively refined as motor learning transforms variable actions into reliable skilled behavior.

neuroscience↗

bioRxiv2026

De novo minibinders targeting RH5 achieve nanomolar inhibition of blood-stage malaria replication

Malaria kills {approx}600,000 people annually despite recently deployed vaccines. The parasite's blood-stage invasion of erythrocytes, driven by the essential, non-redundant interaction between Plasmodium falciparum RH5 and erythrocyte basigin, remains a leading therapeutic target. Here, we designed de novo mini-proteins (minibinders) to occlude this interface. From {approx}1,500 computationally generated designs, we synthesized 18 candidates ranging from 9.3 to 16.9 kDa which were easily expressed in E. coli as soluble, well-folded proteins. Of these, 7 minibinders blocked blood-stage replication of P. falciparum asexual stage parasites with greater potency than {approx}300nM IC50. Three potent candidate minibinders, mb-5, mb-7, and mb-21, were confirmed to bind RH5 via SPR and BLI and block the merozoite-to-ring transition in stage-synchronized parasite progression assays, with CryoEM of minibinder-RH5 complex confirming the predicted binding at RH5-basigin interface. Although pharmacological challenges remain, our data demonstrate that potent inhibitors of parasite growth, rivaling existing antimalarials, can be designed rather than discovered via extensive screening. Our data highlight the potential for artificial intelligence to change the development landscape of new antimalarial therapies.

microbiology↗

bioRxiv2026

Trustworthy super-resolution reconstruction across spatial omics modalities

Sequencing-based spatial omics platforms provide scalable and unbiased profiling of transcriptomic, epigenomic, and isoform-resolved signals, but their spatial resolution remains limited because each capture unit aggregates molecular information from multiple cells. Existing computational enhancement methods reconstruct high-resolution spatial gene expression from spot-level data by integrating histology, but most are designed primarily for spatial transcriptomics, rely predominantly on histological features, and lack appropriate validation strategies, leading to overfitting, spurious spatial patterns, and limited reliability. Here we present spEnhance, a generalizable and trustworthy computational framework for super-resolution enhancement of spot-level spatial omics data. spEnhance integrates histological features with complementary molecular information, including single-cell RNA-seq references and gene co-expression structure, to reconstruct high-resolution spatial molecular profiles. To enable reliable model selection from a single tissue section, spEnhance introduces a count-splitting strategy that generates statistically independent training and validation sets from spot-level measurements. spEnhance further quantifies prediction reliability through predictive residuals, providing an interpretable proxy for spatially resolved uncertainty. Comprehensive benchmarking across multiple tissues, platforms, and molecular modalities demonstrates that spEnhance achieves state-of-the-art accuracy, recovers fine-grained tissue structures, mitigates overfitting, and provides calibrated reliability estimates. Beyond spatial transcriptomics, spEnhance extends to isoform-level, epigenomics, proteomic and metabolomic spatial omics. Collectively, spEnhance establishes a modality-general and trustworthy framework for enhancing spatial omics data, enabling more accurate and reliable investigation of spatially resolved molecular regulation.

bioinformatics↗

bioRxiv2026

A tripartite mechanism licenses RNA polymerase import into the phage nucleus

Nucleus-forming jumbo phages import a phage-encoded non-virion RNA polymerase (nvRNAP) into a proteinaceous nucleus, but how it is selected for import is unknown. Here, we identify an import pathway that couples nvRNAP subunit interactions with phage-encoded import factors. Maximal nvRNAP import requires a novel factor Imp7 (gp166) and the essential import factor Imp1. Imp7 is essential at environmental temperatures (20 C), where impaired nvRNAP import causes a profound loss of middle and late transcription and blocks phage DNA replication. Import of individual nvRNAP subunits also depends on other subunits, consistent with import licensing at the level of an assembled complex. In the absence of middle transcription and DNA replication, the phage nucleus surprisingly still assembles and segregates from the cytoplasm, demonstrating that nvRNAP nuclear localization and activity are not a critical checkpoint for nucleus assembly. Together, these findings define a specialized pathway for nuclear import of the multi-subunit RNAP and reveal that nuclear compartment assembly precedes the transcriptional and replicative programs required for its maturation.

microbiology↗

bioRxiv2026

Parallel Arg/N-degron recognition systems regulate coenzyme A biosynthesis

Coenzyme A (CoA) is an essential metabolic cofactor whose biosynthesis is controlled by pantothenate kinases (PANKs), the rate-limiting enzymes in CoA synthesis. Although CoA production is extensively regulated, whether PANK abundance is controlled by selective protein degradation remains unknown. Here, we identify PANK1{beta} and PANK3 as substrates of the Arg/N-degron pathway. Two mechanistically distinct E3 ligase systems, UBR4-KCMF1 and MKLN1-CTLH, independently converge on a common N-terminal MK Arg/N-degron to promote PANK degradation and restrict CoA biosynthesis. UBR4-KCMF1 recognizes the MK Arg/N-degron through KCMF1 ZZ domain, whereas CTLH complex employs the B30.2-domain proteins RanBP9 and RanBP10 as previously unrecognized Arg/N-degron recognins. Structural, biochemical, and cellular analyses demonstrate that RanBP9 and RanBP10 recognize both Met-X and canonical Arg/N-degrons through their B30.2 domains, using a mechanism distinct from canonical UBR proteins. Together, these findings establish a new branch of the mammalian Arg/N-degron pathway and reveal ubiquitin-dependent control of PANK abundance in CoA homeostasis.

molecular biology↗

bioRxiv2026

Point mutations and complex variants impact gene expression and addiction-related behaviors in Heterogeneous Stock rats

While various variants, including single nucleotide polymorphisms (SNPs), small insertions/deletions, short tandem repeats and structural variants, drive individual genetic differences, their influence on gene expression and complex traits remains unclear. We used short- and long-read sequencing to generate a comprehensive variant catalog in Heterogeneous Stock (HS) rats and performed joint cis-expression quantitative trait loci (cis-eQTL) mapping across five brain regions. We found that non-SNP variants accounted for over 50% of lead regulatory associations, many of which were poorly tagged by nearby SNPs using linkage disequilibrium (LD). Comparison between joint and SNP-only analyses showed that over 46% of shared eQTL genes had a non-SNP lead cis-eQTL, and fewer than half were in strong LD with the corresponding lead eSNP. Linking joint cis-eQTLs to complex trait associations identified mechanisms missed by SNP-only approaches, highlighting the importance of incorporating diverse variant types into genetic studies and providing a foundational resource for the HS rat community.

genetics↗

bioRxiv2026

Emerin loss of function inhibits MyoD-driven differentiation of human iPSCs into skeletal myotubes

Loss of function of the nuclear lamina-associated protein emerin causes Emery-Dreifuss muscular dystrophy (EDMD). Efforts to define emerins essential functions in skeletal muscle have been limited by poor concordance between mouse models and human disease phenotypes. Here, we adapt transgene-driven differentiation of human induced pluripotent stem cells (hiPSCs) into skeletal muscle (iSMs) as a tractable human model for emerin loss of function. We find that EMD knockout (KO) hiPSCs are poorly responsive to combined overexpression of MyoD and Baf60c and produce fewer mature iSMs, indicating that emerin influences muscle differentiation downstream of these differentiation factors. While MyoD acetylation and heterodimerization with E-box proteins are unaffected by emerin loss, MyoD targets including p21 and myogenin are downregulated, and EMD KO cells exhibit impaired cell cycle exit in response to differentiation signals. Transcriptomic analysis of EMD KO iSMs revealed persistent expression of cell cycle genes and decreased expression of terminal muscle differentiation genes. Dysregulated genes do not overlap with lamina-associated domains (LADs) but are instead enriched for targets of polycomb repressive complex 2 (PRC2), which deposits H3K27 trimethylation. Altogether, our data indicate a functional overlap between emerin and PRC2-mediated regulation of terminal muscle differentiation.

cell biology↗

bioRxiv2026

Melittin-induced membrane disruption in hepatocellular carcinoma involves mtROS/GSDME-mediated pyroptosis

Although melittin is classically regarded as a membrane-lytic peptide, it remains unclear whether its membrane-disruptive effects in tumour cells result exclusively from direct peptide-lipid interactions or are partly generated by pyroptosis. Here, we show that melittin markedly suppresses the viability and clonogenic growth of human Huh7 and murine Hepa1-6 HCC cells and rapidly induces characteristic features of pyroptosis, including cell swelling, balloon-like membrane protrusions, plasma-membrane permeabilization and lactate dehydrogenase release. In Hepa1-6 cells, melittin induced prominent GSDME cleavage and accumulation of its pore-forming N-terminal fragment, whereas GSDMD activation was not detected. Quantitative proteomics and ultrastructural analyses further revealed pronounced mitochondrial perturbation after melittin treatment, accompanied by loss of mitochondrial membrane potential and increased mitochondrial reactive oxygen species (mtROS). Importantly, scavenging mitochondrial oxidants with Mito-TEMPO markedly reduced mtROS accumulation, attenuated pyroptotic membrane damage and partially restored cell viability. Collectively, our findings provide, to our knowledge, the first evidence that melittin induces pyroptotic cell death in HCC cells through a mechanism involving mitochondrial oxidative stress and GSDME processing. These results extend the conventional view of melittin as a direct membrane-lytic peptide and uncover an mtROS/GSDME-associated pyroptotic mechanism underlying its antitumour activity in HCC.

cancer biology↗

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↗