Biology subjects

Wang, J.

Publications and source records attributed to Wang, J..

At least 145 records · Page 8Linked to original sources

bioRxiv2026

Oxytocin promotes advantageous inequity preference by modulating the fairness norm enforcement system

Aversion to inequity is innate in humans and constitutes a major contributor to social tension. Such aversion can be triggered either when one receives more (advantageous inequity-AI) or less (disadvantageous inequity-DI) than others. However, it remains unclear whether oxytocin can modulate how humans respond to inequity. The present pre-registered study combined behavioral measures, computational modelling, and fMRI in a novel monetary allocation evaluation paradigm to examine oxytocins modulation of behavioral and neural responses to inequity. Results showed that oxytocin increased perceived fairness and AI allocation choices via enhancing AI preference independent of gain vs. loss contexts, which was further confirmed by model-based results of oxytocin decreasing AI aversion. In parallel at the neural level, oxytocin increased fairness rating-related activity in regions within fairness-related norm enforcement systems involved in norm violation detection, salience processing, and cognitive control. Multivariate-based representational similarity analyses provided further evidence that oxytocin induced distinct representational patterns of inequity in these core regions. These results suggest that oxytocin promoted AI preference possibly by strengthening regulatory control to support the pursuit of economic self-interest, while simultaneously enhancing the salience of AI allocations. Our study provides new insights into behavioral and neural mechanisms through which oxytocin modulates fairness-related behavior.

neuroscience↗

bioRxiv2026

Context-dependent variant interpretation from Mendelian disease to genetic predisposition: a proof-of-concept using LPL

As genomic sequencing evolves beyond rare disease diagnostics toward population screening and precision medicine, clinical variant interpretation is increasingly challenged by variants whose clinical consequences depend on biological context. Current frameworks, including the ACMG/AMP guidelines, generally assign a single classification to each variant regardless of inheritance state or genetic context, potentially failing to communicate context-dependent clinical consequences. Here, we address this issue using loss-of-function variants in LPL as a uniquely informative model system in which residual physiological LPL activity can be directly quantified in vivo. By systematically integrating published biallelic LPL genotypes, physiological measurements, functional studies, and clinical phenotypes, we identified a biologically meaningful transition at approximately 10% residual physiological LPL activity. Activity below this level was predominantly associated with classical childhood-onset familial chylomicronemia syndrome (FCS), whereas higher activity was associated with phenotypic attenuation and modifier-dependent clinical expression. Furthermore, heterozygous loss-of-function variants exhibited an estimated penetrance of 5-7% for severe hypertriglyceridemia. We therefore propose a context-dependent framework in which biallelic complete- or near-complete loss-of-function genotypes are interpreted as causative for FCS, whereas heterozygous variants are interpreted as predisposing to severe hypertriglyceridemia while retaining recognition of FCS carrier status. Together, our findings demonstrate that clinical variant interpretation should integrate available biological context--including, where relevant, allelic configuration, residual biological function, and penetrance--rather than rely on the intrinsic molecular consequence of the variant alone. More broadly, this framework provides a conceptual model for interpreting variants across the continuum from Mendelian disease to genetic predisposition in the era of precision medicine.

genetics↗

bioRxiv2026

A neurofunctional signature of romantic love predicts rewards on social media, effects of oxytocin and drug-cue reactivity

Romantic love is a selective motivational state supporting pair bonding, yet its neural representation and relation to other affiliative-rewarding experiences remain unclear. Across eight fMRI studies (n=420) spanning naturalistic affiliative-reward, pharmacological and addiction-relevant experiments, we developed and evaluated multivariate whole-brain decoders of romantic love and friendship. Both signatures engaged mesocorticolimbic reward systems and the precuneus, yet were dissociable at the whole-brain level and in their recruitment of social-cognitive systems. The love signature generalized to reward on social media, was specific to positive valence, and did not track sweet-taste or monetary reward, indicating a distinct social-affiliative reward representation. Oxytocin selectively increased love - but not friendship - signature reactivity to the romantic partner. In an independent drug-cue-reactivity dataset, the love signature specifically identified heavy cannabis users. These findings establish a dissociable neurofunctional signature of romantic love in conserved bonding circuits that extend to digital affiliation and are co-opted in addiction.

neuroscience↗

bioRxiv2026

SAMP V2: A novel stacking ensemble learning model for antimicrobial peptides identification based on augmented split amino acid composition with biochemical-sequence-order information

Antimicrobial resistance reduces the effectiveness of conventional antibiotics and has become a major global health threat, highlighting the need for new anti-infective agents. Antimicrobial peptides (AMPs), a diverse class of innate immune effectors with broad-spectrum antimicrobial activity, are promising candidates for combating drug-resistant infections. Identifying AMPs by wet-lab experiments, however, remains costly and time-consuming, creating a strong demand for computational identification methods. Our recently developed method, SAMP, captures region-specific residue distributions based on proportionalized split amino acid composition. However, SAMP might ignore key biochemical information and sequence order information. Here we present SAMP V2, a stacking ensemble learning framework based on biochemical and sequence-order information augmented split amino acid composition (BIA-SAAC), which extends SAMP by integrating pseudo-amino acid composition features with biochemical and sequence-order information into split peptide regions. Specifically, each peptide is divided into N-terminal, middle, and C-terminal regions, and pseudo amino acid composition is calculated within each region. Benchmarking tests on six independent test datasets, SAMP V2 outperformed multiple state-of-the-art models, including AMPpred-MFA and iAMP-Attenpred, in terms of accuracy, MCC, G-measure and F1-score. Given its high and robust performance, SAMP V2 could significantly accelerate the discovery of next-generation antimicrobial therapeutics for addressing the global threat of multidrug-resistant pathogens.

bioinformatics↗

bioRxiv2026

The Maine Coon Cat Harboring the MYBPC3-A31P Mutation: A Genotype-Stratified Phenotypic Characterization of Hypertrophic Cardiomyopathy

Abstract Background: Most animal models of HCM are mouse-based, but the thin interventricular septum in mice makes it difficult to clearly distinguish pathological hypertrophy, which introduces substantial errors and constrains basic HCM research. Cats develop HCM spontaneously, and the common MYBPC3-A31P variant in cats is homologous to human mutations in both genetics and pathology, with a larger body size that makes them suitable as large-animal models. This study examines how heterozygosity or homozygosity for the p.A31P mutation (c.91G>C) in the MYBPC3 gene affects the phenotype and severity of HCM in affected cats, with the aim of establishing an ideal large-animal model for clinical risk stratification and precision diagnosis and treatment of human HCM. Methods: Forty-nine Maine Coon cats were enrolled and stratified into homozygous mutant (HOM, n=8), heterozygous mutant (HET, n=26), and wild-type (WT, n=15) groups. All cats underwent echocardiography, blood pressure measurement, physiological assessment, hematological and biochemical analyses, and cross-species sequence conservation analysis. Results: No significant differences in baseline characteristics including age and body weight were observed among groups (P>0.05). HOM cats exhibited significantly higher left ventricular outflow tract pressure gradients and greater basal septal thickness compared to WT cats (P<0.05), with HET cats showing intermediate values. Analysis of hematological and serum biochemical parameters revealed no evidence of systemic inflammation or hepatic injury. Sequence conservation analysis confirmed that the A31 residue is highly conserved across mammalian species. Conclusions: This study provides a phenotypic characterization of Maine Coon cats carrying the MYBPC3-A31P mutation, revealing marked gene-dose effects on cardiac structure and function, with homozygous individuals exhibiting more severe phenotypic features. This model serves as a large-animal translational platform that not only clarifies genotype-phenotype correlations but also supports risk stratification and precision therapeutic strategies in human HCM. Its spontaneous nature and genetic homology to human disease make it particularly valuable for bridging preclinical findings to clinical application.

genetics↗

bioRxiv2026

Unique CD8+ T Cell Populations Expand during ART and Predict Delayed HIV-1 Rebound

Antiretroviral therapy (ART) suppresses HIV-1 replication but does not eliminate the latent reservoir, resulting in viral rebound with variable kinetics after treatment interruption. How the immune cell states established during ART influences timing of rebound is not fully understood. In this study, we analyzed 111 participants across multiple cohorts, with 188 single-cell multiomic samples generated and integrated for joint analysis. Longitudinal profiling of peripheral blood mononuclear cells from individuals with acute HIV-1 infection on ART, spanning early infection through sustained therapy and pre-analytical treatment interruption, revealed that time to viral rebound was driven not by global changes in immune composition but by dynamic transcriptional programs within CD8+ T cells. During ART, there was a dramatic expansion of a unique cluster of poised naive CD8+ T cells, with a distinct immune state positioned upstream of stem-like memory CD8+ T cells along a cell differentiation continuum. The differential abundance of this poised naive CD8+ T cell population was enriched in participants with delayed rebound and showed strong predictive power for discriminating time to rebound. Mechanistically, the poised naive CD8+ T cells exhibited features of a precursor phenotype of stem-like memory CD8+ T cells, and showed activation of the TNF-NF-{kappa}B signaling pathway and increased chromatin accessibility at AP-1 motifs. Notably, both poised naive CD8+ T cells and stem-like memory CD8+ T cells were consistently enhanced during ART in both acute and chronic infection. In participants who received investigational therapeutic vaccination, the dominant predictive signal shifted downstream along the differentiation trajectory, with stem-like memory CD8+ T cells emerging as the primary determinant of delayed rebound. Together, these findings identify a dynamic CD8+ T cell state continuum as a central determinant of HIV-1 rebound, even in the absence of antigen-specificity, where ART establishes a predictive poised naive state that can be further leveraged by vaccination to enhance protective stem-like memory responses.

immunology↗

bioRxiv2026

NIR-II squeezed light-field microscopy enables high-speed volumetric imaging of deep-tissue dynamics in vivo

High-speed three-dimensional imaging in scattering biological tissues remains challenging because volumetric microscopy generally requires scanning, whereas snapshot light-field approaches divide limited detector pixels among multiple views. This constraint is particularly severe in the second near-infrared window (NIR-II), where InGaAs cameras have small sensor formats and high detector noise. Here we introduce NIR-II squeezed light-field microscopy (NIR-II SLIM), which optically rotates and compresses multiple perspective views before detection, allowing efficient use of camera pixels while retaining complementary spatial information for three-dimensional reconstruction. NIR-II SLIM acquires up to 600 volumes s-1 with a reconstructed lateral sampling grid of 512 x 512 pixels. We use this method for label-free imaging of cardiac dynamics in pigmented late-larval zebrafish, resolving chamber deformation and millisecond-scale atrioventricular-valve motion, and for NIR-II fluorescence imaging of vascular and lymphatic transport in mice. NIR-II SLIM provides a detector-efficient approach for high-speed volumetric imaging of rapid biological dynamics in scattering tissues.

bioengineering↗

bioRxiv2026

Antibiotic condensates activate magnesium signaling to drive drug tolerance

The spatial distribution of small molecules within cells shapes their biological activity, yet these distributions are generally assumed to be governed passively by reaction-driven electrochemical gradients. Here we show that aminoglycoside antibiotics actively control their own subcellular organization by undergoing phase separation with RNAs. Combining in vitro reconstitution, bacterial assays, and molecular dynamics simulations, we discovered that aminoglycosides coacervate with RNA through multivalent electrostatic interactions, displacing and releasing RNA-bound Mg2+. This condensate-dependent Mg2+ release remodels the cytosolic labile Mg2+ pool and activates magnesium signaling. This effect dampens the magnesium-starvation regulation, sustains ribosome activity, and shifts the cellular electrochemical state, promoting bacterial fitness. Because condensation occurs only above a defined concentration threshold, it generates a non-monotonic dose-response in which higher antibiotic concentrations paradoxically enhance bacterial survival. This antibiotic condensate-dependent Mg2+ signaling confers tolerance to multiple ribosome-targeting antibiotics simultaneously even in cells lacking resistance gene, while condensate dissolution restores antibiotic efficacy. Our findings establish antibiotic-driven phase separation as a previously unrecognized mechanism to encode cellular signaling and identify antibiotic condensates as a distinct functional unit underlying drug tolerance.

microbiology↗

bioRxiv2026

Architectural trade-offs between environmental stability and genomic redundancy reveal divergent pneumoviral entry strategies

Human respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) exhibit distinct seasonal epidemiology, with RSV circulating in early autumn and hMPV peaking in midwinter, yet the structural basis for this niche partitioning remains undefined. Here, we integrate in situ cryo-electron tomography and functional virology to decode the architectural logic governing their entry dynamics. RSV employs a matrix (M)-regulated prefusion F (pre-F) organization, partitioning trimers into stabilizing hexagonal superlattices and fusion-competent pools to maintain superior thermotolerance. By contrast, hMPV compensates for its intrinsically unstable, monomeric pre-F with extreme ribonucleoprotein polyploidy, packaging [~]4-fold more genome equivalents to ensure productive infection. Fusion events localize exclusively to M-depleted, non-arrayed membrane regions, establishing a spatial checkpoint for activation. These findings reveal a conserved trade-off between environmental resilience and genomic redundancy that dictates divergent pneumoviral entry strategies, explaining their distinct seasonal ecological niches.

microbiology↗

bioRxiv2026

A Principled Statistical Framework for Analyzing Spatial Patterns in Spatially Resolved Multi-Omics

Emerging spatial multi-omics technologies enable the profiling of molecular variation within its tissue context, yet existing methods for identifying spatially variable features lack principled approaches to experimental design and cross-sample inference. Here, we present STORM, a principled Statistical TOol for spatially Resolved Multi-omics, for rigorously analyzing spatial patterns in spatial multi-omics research. STORM incorporates a robust and efficient nonparametric test that quantifies local deviations in molecular feature measurements to detect spatial dependence across transcriptomic and proteomic data. It further estimates an interpretable spatial effect size, supports power calculations for both spatial locations and biological replicates, and enables formal group-level comparisons. In several simulated and experimental spatial multi-omics case studies, STORM demonstrates reliable performance in detecting spatial structures while offering quantitative support for study design decisions. Overall, STORM provides a principled statistical framework that unifies spatial hypothesis testing, effect size estimation, power analysis, and experimental design for spatial multi-omics data.

bioinformatics↗

bioRxiv2026

Extracellular vesicle-mediated suppression of macrophage STING signaling promotes immune dysfunction in dedifferentiated liposarcoma

BackgroundDedifferentiated liposarcoma (DDLPS) is characterized by abundant immune cell infiltration yet derives limited benefit from immune checkpoint blockade and stimulator of interferon genes (STING) agonist-based strategies, suggesting tumor-mediated suppression of antitumor immunity. Tumor-associated macrophages are the most abundant immune populations in DDLPS, but the factors regulating their function remain incompletely understood. MethodsExtracellular vesicles (EVs) were isolated from two DDLPS cell lines and serum from 16 DDLPS patients and 13 healthy donors. EVs impact on cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) -induced macrophage activation was assessed by cytokine secretion, surface markers, functional assays and macrophage-T-cell coculture. Proteomics was performed in EV-treated and EV-untreated macrophages from three donors. Pathway and protein interaction analyses were integrated with The Cancer Genome Atlas (TCGA) DDLPS transcriptomic and survival data. ResultsWe show that EVs released by DDLPS cells suppress macrophage responsiveness to classic STING agonist cGAMP. EVs derived from DDLPS attenuated cGAMP-induced expression of type I interferon-associated cytokines and chemokines, reduced IFN-{beta} secretion, and impaired phosphorylation of STING, TBK1 and IRF3. Functionally, DDLPS EV exposure shifted macrophages toward an immunoregulatory phenotype, restrained phagocytic activity, and attenuated macrophage-dependent T-cell proliferation while promoting T-cell exhaustion. Proteomic profiling revealed extensive macrophage reprogramming characterized by suppression of STING-associated signaling, antigen processing and presentation associated pathways and proteins targeted by miR-16-5p. Consistent with these findings, STING expression was associated with prolonged overall survival in DDLPS, while reduced expression of miR-16-5p target proteins was associated with attenuated STING pathway activity and immunostimulatory macrophage signatures. ConclusionsThese findings identify EV-mediated suppression of macrophage STING signaling as a mechanism of immune dysfunction in DDLPS and provide a framework for understanding immune resistance in this disease.

cancer biology↗

bioRxiv2026

Cellular senescence is associated with age-related loss of liver zonation and hepatocyte function

The liver is organized into tightly regulated zones with distinct metabolic functions but zonation erodes with age. Cellular senescence contributes to aging and liver diseases, however, its impact on aging biology is ill-defined. As part of The Cellular Senescence Network Consortium, we used multiple spatial transcriptomics approaches (GeoMx, Visium, CosMx) with snRNA-seq to profile senescence signatures, zonation markers, and metabolic pathways in livers from wild-type (WT) mice of multiple ages. We observed a loss of canonical zone signatures in aged mouse livers characterized by "expansion" of midlobular (zone 2) marker gene expression, accompanied by diminished expression of zone 3 marker genes by middle-age (18 months), indicative of loss of cell identity. Multiple analytic approaches identified distinct age-, zone- and sex-specific senescence signatures, which were significantly associated with zonation markers changes. This was recapitulated in Ercc1 mutant models of accelerated senescence, supporting a causal role of senescent cells in liver aging. A "no-zone" hepatocyte-like cluster expanded with age and with the strongest Senescence-Associated Secretory Phenotype (SASP) profile. Gene expression profiles from senescent hepatocytes implicate decreased WNT signaling and increased BMP as contributing to age-related loss of zonation. Together, these data elucidate the role of senescent cells in driving aging biology in non-diseased liver through disruption of cell:cell signaling and the loss of metabolic and cell identity gene expression necessary for hepatocyte function.

cell biology↗

bioRxiv2026

First detected incursions of avian influenza H5N1 clade 2.3.4.4b into mainland Australia from the Southern Ocean

High pathogenicity avian influenza H5N1 clade 2.3.4.4b has caused a panzootic of devastating impact to poultry and wildlife globally. The Australian continent and broader Oceania until recently remained the last major region without confirmed detections. Here we report the first H5N1 clade 2.3.4.4b detections from two live seabirds - a brown skua and a southern giant petrel - found on the south coast of Western Australia in June 2026. Virus genome sequencing showed that both viruses were most closely related to H5N1 viruses detected recently on sub-Antarctic islands in the Southern Indian Ocean. In time-calibrated phylogeographic analyses, both viruses sampled in Western Australia clustered with viruses from Heard Island, a sub-Antarctic external territory of Australia. Ancestral location reconstruction also identified Heard Island as the most probable source location, although unsampled intermediate locations cannot be excluded. The two Western Australian detections were estimated to be independent incursions from Heard Island, rather than local transmission on mainland Australia. There was no evidence of reassortment with endemic avian influenza viruses in Australia, and both virus sequences retained key avian-like genetic markers and lacked known substitutions for reduced antiviral susceptibility. These detections revealed a Southern Ocean pathway of recurrent H5N1 incursions into Australia, highlighting the risk of potential establishment on the mainland and the need for heightened surveillance and rapid, nationally-coordinated, virus genomic characterisation.

microbiology↗

bioRxiv2026

Genomic and eco-geographic features of locally adapted inversions in wild sunflowers

Chromosomal inversions are increasingly recognized as important drivers of local adaptation and ecological divergence because they suppress recombination and maintain adaptive allele combinations despite ongoing gene flow. However, the eco-evolutionary conditions favouring the establishment of such indirectly adaptive inversions, as well as the genomic features that distinguish them from other inversions remain incompletely understood. In this study, we investigated these questions in a wild sunflower system comprising two species: Helianthus debilis and Helianthus praecox, which exhibit diverse ecotypes and varying degrees of geographic overlap across Texas and Florida in the USA. To resolve the evolutionary relationships between and within these species, and identify potentially adaptive inversions, we generated haplotype-resolved reference assemblies and integrated comparative and population genomic analyses. We identified three major genetic clusters that only partially corresponded to taxonomic classifications. We further detected 156 inversions across the genome, 11 of which showed signatures consistent with a role in local adaptation. Notably, nine of the 11 putatively adaptive inversions were found in sympatric Texas populations. Together with a similar enrichment of inversions in genome assemblies from sympatric versus allopatric populations, our results suggest that inversions are more likely to evolve in heterogeneous environments with ongoing gene flow than in allopatry. Lastly, locally adaptive inversions were generally larger, contained more genes, and showed greater sequence divergence between haplotypes than other types of inversions. Our findings provide empirical support for the role of gene flow in promoting inversion establishment and identify genomic characteristics associated with indirectly adaptive inversions.

evolutionary biology↗

bioRxiv2026

Mechanical Activation of Piezo1 by Virus-like Nanospikes to Potentiate STING-driven Macrophage Reprogramming

Mechanotransduction plays a fundamental role in regulating immune cell function, yet how engineered virus-like nanospikes engage mechanosensitive signaling pathways to modulate innate immunity remains poorly understood. Here, we report virus-like nanotopography as a previously unrecognized regulator of Piezo1-mediated mechanotransduction in macrophages using virus-like mesoporous silica nanoparticles (VLPSi) with tunable rigid nanospike lengths. We demonstrate a direct structure-activity relationship between nanospike geometry and Piezo1-dependent Ca{superscript 2}+ influx, with longer nanospikes inducing significantly greater intracellular Ca{superscript 2}+ signaling. Building on this mechanistic insight, we developed biomimetic cancer cell membrane (CM)-coated, MSA-2-loaded VLPSi nanoparticle (CM/MSA-2@VLPSi) and investigate the combination of nanospikes-activated Piezo1 with STING signaling and CM antigens presentation in macrophage immune reprogramming. The resulting biomimetic nanoparticles robustly activate the STING-TBK1-IRF3/NF-{kappa}B axis, increase IFN-{beta} and pro-inflammatory cytokine production, and promote macrophage polarization toward M1 phenotype in a spike-length-dependent manner. Collectively, the present study provides a biomimetic strategy for enhancing the M1 polarization of macrophage through the coordinated regulation of mechanical, inflammatory, and antigenic signals.

immunology↗

bioRxiv2026

Allergen-responsive T helper type 2 cells revealed by high-dimensional profiling in allergen challenged human airways

Asthma is a chronic inflammatory disease affecting over 300 million people worldwide. This disease has multiple underlying etiologies, and a major endotype of asthma is characterized by cellular and molecular signatures of type 2 (allergic) inflammation. In this study we conducted bronchoscopies with airway segmental allergen challenge in allergic asthmatics to dissect airway responses to allergen. Using mass cytometry and single-cell RNA sequencing, we characterized with high resolution the airway immune landscape before and after allergen challenge and the heterogeneity present between subjects. This heterogeneity generally falls along a type 1/ type 2 axis. In type 2 high individuals, we identified allergen-reactive Th2 cells by using TCR sequences to barcode clonal T cell populations in single-cell genomic and activation-induced marker expression assays. These potentially pathogenic Th2 cell clones were present systemically and expanded following allergen challenge, connecting local lung inflammation to systemic clonal Th2 cell dynamics. Th2 cell airway ingress was coordinated with myeloid cell expression of T cell chemoattractants including CCL17 and CCL22. This study provides insight into the molecular and cellular components of allergen-induced tissue inflammation in asthma. Deeper resolution of the T cell response to aeroallergens may inform novel diagnostic and therapeutic strategies for asthma and other allergic airway diseases.

immunology↗

bioRxiv2026

A nanoconfined liquid-in-ice aqueous regime for time-stretched single-molecule dynamics

The experimentally observable dynamical landscape of biomolecules is fundamentally shaped by rapid thermal motions of the surrounding aqueous environment. Although lowering temperature could expand this observable landscape, the liquid-solid phase transition of water has long prevented real-time single-molecule measurements into deeply subzero aqueous environments. Here we show that nanoconfinement within a solid-state nanopore overcomes the fundamental limitation imposed by bulk water freezing, spontaneously stabilizing a persistent liquid-in-ice environment that remains electrically accessible despite surrounding electrolyte crystallization. This aqueous environment creates a time-stretched dynamical regime, extending molecular translocation timescales by up to [~]400-fold and revealing previously inaccessible single-molecule dynamics. These findings establish a new low-temperature aqueous regime for real-time single-molecule measurements, opening new opportunities to investigate biomolecular dynamics across previously inaccessible timescales and extreme aqueous environments.

biophysics↗

bioRxiv2026

Oncogenic transformation proceeds through a transient state of cellular plasticity constrained by lineage-specific barriers.

Normal tissues frequently harbor oncogenic mutations without progressing to cancer, but the cellular basis of this resistance remains poorly defined. We asked whether transformation requires selection of a rare, permissive state within normal-like cell populations. Dermal fibroblasts and mammary epithelial cells responded uniformly to combined HRAS-G12V expression and p53 disruption. Cellular barcoding revealed no loss or enrichment of clones during morphological transformation, arguing against clonal selection as the primary driver of neoplastic reprogramming. Instead, all transduced cells underwent an early, shared transcriptional transition characterized by loss of differentiation markers, induction of RAS-associated and inflammatory programs, activation of alternative-lineage signatures, increased single-cell entropy, and chromatin decondensation. These changes were transient: entropy and chromatin accessibility subsequently declined, and some cells moved toward the control transcriptional state, whereas others stabilized in altered states. The two lineages followed distinct trajectories. Fibroblasts showed greater initial transcriptional plasticity but subsequently reverted more strongly toward the normal state, whereas epithelial cells changed more gradually and continued to diverge from it, suggesting a stronger barrier to transformation in the mesenchymal lineage. Thus, oncogenic perturbation initiated reprogramming throughout the population but did not uniformly produce a stable transformed state. Together, these findings support a model in which normal-like cells tolerate oncogenic mutations not because most cells fail to respond, but because a p53-independent, cell-intrinsic barrier limits the stabilization of malignant transformation following a transient period of heightened plasticity. This framework may facilitate the identification of mechanisms that constrain tumor initiation.. Significance StatementLoss of TP53 and activation of oncogenic RAS are common drivers of human cancer and frequently coexist in the same tumor. Nevertheless, these alterations often fail to induce malignant transformation. To investigate why, we introduced both alterations into fibroblasts and epithelial cells and tracked their responses using clonal barcoding, single-cell transcriptomics, and phenotypic assays. Initially, cells uniformly reprogrammed their transcriptional and chromatin states without detectable clonal selection. They then diverged: some reacquired transcriptional profiles resembling those of the original normal cells, whereas others became transformed. Thus, normal cells resist transformation through an intrinsic barrier that operates despite p53 disruption. Because this transition is transient and reversible, it provides a tractable window for studying the earliest stages of cancer development.

cancer biology↗