Biology subjects

Wu, X.

Publications and source records attributed to Wu, X..

At least 19 recordsLinked to original sources

bioRxiv2026

Transient diquat inclusion and carboxylate-rim binding to carboxymethyl-α-cyclodextrin: a molecular dynamics study

Rotaxane-based supramolecular scavengers have shown promise for diquat detoxification, but the relative contributions of cavity inclusion and interactions at the macrocycle rim remain unclear. We used all-atom molecular dynamics simulations in explicit water to examine diquat binding to carboxymethyl--cyclodextrin (CM--CD) and the behaviour of CM--CD on a poly(ethylene glycol) axle. Five systems were simulated: host-guest mixtures containing a neutral host (pH 3 model) or a hexaanionic host (pH 10 model), an explicitly constructed inclusion complex, and two-macrocycle pseudopolyrotaxanes initiated from pre-threaded or random configurations. The total production sampling was approximately 4.5 s. A relaxed threaded geometry was obtained, with the diquat centroid 0.03 nm from the cavity axis and 9 of its 14 heavy atoms within the defined cavity cylinder. The guest left this geometry within the first 50 ps sampling interval and subsequently remained associated mainly with the carboxylate rim. During 1.2 s of unbiased sampling of the hexaanionic host-guest system, only 5 of 802 guest-frame pairs contained a guest atom within the cavity cylinder, and all of these events occurred near a portal. Rim contacts were more frequent: each guest formed an average of 3.8-4.4 host heavy-atom contacts per frame, with 12.8-14.0% of guest heavy atoms in contact with the host. These interactions were substantially reduced for the neutral host. Sodium ions remained predominantly solvent-separated from the carboxylates. In the pseudopolyrotaxane simulations, pre-threaded rings satisfied the threading criterion in 1.6-8.4% of frames, and no spontaneous threading was observed from random configurations during 1 s. Under the simulated conditions, CM--CD therefore bound diquat primarily through dynamic rim interactions rather than persistent cavity inclusion.

biochemistry↗

bioRxiv2026

Transcriptomic similarity guides source selection beyond tissue lineage

Perturbational and pharmacogenomic maps remain incomplete across cellular contexts, leaving a practical source-selection question before response prediction: which measured context should inform an unmeasured target? We tested whether baseline transcriptomic similarity ranks candidate sources for held-out targets, separating source ranking from aggregation and transfer-uncertainty decisions. Similarity tracked matched-response concordance in LINCS and GSE306429; selected-source utility exceeded random expectation in GSE306429 ({Delta}U = 0.034; P = 0.002) and in exact-time-matched MIX-Seq tasks ({Delta}U = 0.046). In the full GDSC2 analysis, the drug-balanced ranking effect was 0.359 (95% CI, 0.337-0.383). In a lineage-restricted analysis of the same eligible subset, within-tissue ranking remained positive at 0.130 (95% CI, 0.119-0.141), retaining 35.8% of the same-subset global effect (0.363; 95% CI, 0.342-0.386). Global transcriptomic top-1 selection exceeded tissue-matched random selection by 0.154 (95% CI, 0.143-0.164). Thus, baseline similarity adds source-ranking information after tissue matching in the evaluated setting, while ranking alone does not resolve whether to aggregate sources or how to quantify transfer uncertainty.

bioinformatics↗

bioRxiv2026

Glioblastoma Hijacks Astrocyte Ribosomes to Feul Malignancy through Translatome Reprogramming

Cancer cells demand elevated protein synthesis, yet whether they directly requisition ribosomes from microenvironmental host cells remains unknown. Here, utilizing in vivo and in vitro ribosome tracing, we show that glioblastoma (GBM) cells hijack ribosomes from surrounding astrocytes via tunneling nanotube-like structures. This transfer supports GBM malignancy in both murine and patient derived models. Single cell RNA sequencing along with whole cell and cell source traceable ribosome profiling reveal that transferred ribosomes are translationally active and selectively reprogram the tumor translatome, prioritizing cell-cycle genes and driving interferon-response-mediated immune evasion. Through a genome wide CRISPR screen, we identify INTS12 as a critical mediator of ribosome transfer. Genetic inactivation or AAV-mediated knockdown of INTS12 impairs ribosome uptake, curbs translation and proliferation, remodels the immunosuppressive landscape, and extends survival in orthotopic models. Our study uncovers a parasitic translation-support mechanism in GBM, establishing INTS12 as a therapeutic vulnerability to disrupt ribosome transfer.

cancer biology↗

bioRxiv2026

Therapeutic Targeting of the TGFβ Pathway Augments CAR T cell Efficacy in Glioblastoma

Chimeric Antigen Receptor (CAR) T therapy represents a promising approach to improve outcomes for patients with high-grade gliomas (HGG). Recently, we reported on a phase I trial assessing the local delivery of CAR T cells for HGG. Despite notable clinical benefits to CAR T treatment, clinical responses were highly variable between patients. Here we report that TGF{beta} levels in the tumor cavity fluid (TCF) decreased following CAR T treatment and that lower TGF{beta} levels positively correlated with improved patient responses to CAR T therapy. Patients were stratified based on baseline TGF{beta} levels in the TCF before CAR T therapy. Single-cell RNA sequencing of patient tumor tissues identified significant upregulation of genes associated with angiogenesis and epithelial-to-mesenchymal transition, and IL6 and SLRP pathways in TGF{beta} high patients, while IL2, chemokine signaling and NK-mediated cytotoxicity pathways were significantly upregulated in TGF{beta} low patients. Preclinically, we demonstrated that systemic treatment with a TGF{beta}R1 inhibitor markedly improved CAR T efficacy in syngeneic murine glioma models, correlating with changes in the tumor microenvironment (TME) that benefited both endogenous and CAR T cell phenotype and trafficking. Further studies demonstrated that inhibition of the TGF{beta} pathway in CAR T cells, through either TGF{beta}R1 inhibitor or CRISPR/Cas9 knockout of TGF{beta}R2 (TGF{beta}R2-KO), significantly enhanced CAR T cell expansion and antitumor activity. Collectively, our clinical and preclinical findings strongly indicate that targeting the TGF{beta} pathway could significantly enhance CAR T therapy for patients with high-grade gliomas.

cancer biology↗

bioRxiv2026

A myosin filament cage drives multiple aspects of mitosis unique to cells dividing within a polarized epithelium

Here we used mouse intestinal organoids expressing endogenously tagged myosin to clarify its roles in those aspects of mitosis unique to division within a polarized epithelium. Super-resolution imaging of rounded metaphase cells revealed a never-before-seen myosin filament cage that only forms in this context, exhibits apical/basal polarity, maintains roundness by resisting surrounding tissue pressure, and repositions laterally to form a contractile ring predetermined to drive planar division and asymmetric cleavage furrow progression. Attenuating cage function inhibited interkinetic nuclear migration and metaphase cell rounding, causing the division plane to shift from planar to orthogonal and resulting in organoids possessing multiple lumens. Following the apical domain marker ZO-1 showed that the new lumens originate at the interface between stacked daughter cells created by orthogonal divisions. We conclude that cells dividing within polarized epithelia create a myosin filament cage designed specifically to drive those aspects of cell division unique to their context.

cell biology↗

bioRxiv2026

UnibindMD: An Integrated AI and Molecular Dynamics Platform for De Novo Peptide Binder Design

UnibindMD is an integrated computational platform that turns a protein target into validated de novo peptide-binder leads by chaining five orthogonal stages: deep-learning binder design (BindCraft), independent structure prediction and confidence assessment (AlphaFold3-class / Boltz-2), binding-affinity prediction (PRODIGY), developability profiling (ADMET-AI), and all-atom molecular-dynamics validation (GROMACS/CHARMM36m). The platform targets a persistent weakness of single-method design, namely that confidence metrics such as ipTM and pLDDT are self-consistent but not independent, so a design predicted to be confident may still be physically unstable. We demonstrate UnibindMD on three Andes virus (ANDV) intervention points, a setting of urgent need given a case fatality rate approaching 40%, a 2026 MV Hondius cruise-ship outbreak (13 cases, 3 deaths), and no approved therapeutic: the Gn-Gc glycoprotein complex (viral entry), the L-protein endonuclease domain (cap-snatching; PDB 6Q99), and the host VEGFR2 kinase (vascular permeability; PDB 2XIR). Across nine BindCraft campaigns (4,475 trajectories) the platform produced 464 accepted binders (acceptance 2.3-67.6%), with top candidates reaching ipTM = 0.94 and pLDDT = 0.94. The three selected leads were predicted by PRODIGY to bind with G = -9.1 to -11.1 kcal mol-1 (Kd 8 nM-0.2 M), and all three remained stably bound over 100 ns of unrestrained molecular dynamics (receptor CRMSD [≤] 2.6 [A], retained peptide fold, [≥] 99.5% of frames preserving more than half of the initial interface contacts, and 75-82% helical content). ADMET-AI flagged the expected poor oral bioavailability of large polar peptides and no PAINS alerts. By converting single-prediction design into a cross-validated, dynamics-tested workflow, UnibindMD provides a rapid, scalable, and generalizable path from any protein target to validated peptide-binder candidates.

bioinformatics↗

bioRxiv2026

FAK Dependent Focal Adhesion Remodeling Drives Arsenical Vesicant Induced Skin Injury

Chemical vesicants cause severe skin injury characterized by inflammation, tissue destruction, and separation of the epidermis from the underlying dermis. However, the molecular mechanisms linking vesicant exposure to the disruption of epidermal adhesion remain poorly understood. Phenylarsine oxide (PAO), a commonly used lewisite analog, is a potent inducer of cutaneous blistering and a strong inhibitor of protein tyrosine phosphatases. Here, we show that PAO rapidly induces focal adhesion kinase (FAK) phosphorylation and downstream signaling in epidermal keratinocytes, accompanied by accelerated focal adhesion disassembly and molecular turnover. Genetic ablation of FAK stabilizes focal adhesions and markedly suppresses PAO-induced adhesion remodeling. Importantly, epidermal-specific deletion of FAK protects mouse skin from PAO-induced microvesication, inflammatory responses, and cell death. Pharmacological inhibition of FAK similarly preserves focal adhesion stability in vitro and attenuates PAO-induced skin injury in vivo, phenocopying the protective effects of genetic FAK deficiency. Together, our findings identify aberrant FAK activation as a critical signaling event linking arsenical vesicant exposure to focal adhesion failure and skin vesication, and suggest that targeting FAK may provide a therapeutic strategy to mitigate chemical vesicant-induced skin injury.

cell biology↗

bioRxiv2026

Why 627 BindCraft2 trajectories produced zero binders

De novo protein-binder design with deep-learning pipelines such as BindCraft2 has been reported to deliver experimental success rates of 10-100% against well-structured targets. Here we report the opposite tail: two BindCraft2 (v1.0.3) cyclic-peptide campaigns targeting the shallow K81 lactylation epitope of human lactate dehydrogenase A (LDHA) produced zero accepted designs from 627 trajectories. Retrospective analysis of the campaign artifacts shows that 97% of trajectories were rejected at the first ("screen") stage by a single confidence gate (binder pLDDT < 0.60), not because the interface failed to form: the interface metric i_pTM reached 0.69 and the target was confidently folded (pTM = 0.90). Critically, the mid-project decision to disable the "lysine trick" (forced_targeting) was backwards -- that feature moved 17 trajectories past the screen versus 1 without it, and raised interface confidence. We document two preset/metric mismatches that set a macrocycle campaign up to fail regardless of budget: a screen-stage pLDDT gate calibrated for globular binders, and a binder_contacts loss that is degenerate for short peptides. We describe the exact artifacts BindCraft2 writes and distill them into a reusable post-mortem protocol, and we include a complete how-to-use-BindCraft2 guide so the failure analysis also serves as a tutorial.

bioinformatics↗

bioRxiv2026

Nicotinic modulation of frontal circuits breaks the cycle of attentional performance failure

The ability to recover from consecutive failures is critical for adaptive behavior, yet the underlying neural mechanisms remain unclear. In vivo screening of neuromodulatory inputs onto frontal-sensory cognitive control circuit identified a frontal cholinergic mechanism that counteracts lapses in attentional engagement following repeated errors. Cholinergic input from the basal forebrain to the frontal cortex was necessary and sufficient to restore performance after errors, specifically after repeated-but not after a single-failure. This effect was mediated by 7-nicotinic acetylcholine receptors in frontal-sensory cortical neurons, which sustained cortical circuit activity and frontal-sensory cortical oscillations during attentional engagement. Targeted optogenetic activation of these frontal-sensory neurons disrupted cycles of attentional performance failure and reinstated behavioral performance in a mouse model of neuropsychiatric syndrome. These findings reveal a circuit-level mechanism of cholinergic control that confers cognitive resilience under conditions of persistent failure.

neuroscience↗

bioRxiv2026

Large-scale all-optical electrophysiology links excitability and connectivity to circuit dynamics in vivo

To understand neural computations, one would like to relate circuit dynamics to neuronal intrinsic excitability and connectivity. We developed tools for large-scale all-optical electrophysiology and applied them to approximately 300 layer 1 interneurons per mouse in vivo. Across two mice, targeted stimulation and voltage imaging identified 1,469 inhibitory and 5,752 directed electrical connections among 14,312 tested pairs. Presynaptic identity was more strongly associated with inhibitory post-synaptic potential (IPSP) amplitude, whereas postsynaptic identity was more strongly associated with IPSP decay time. Gap junctional and synaptic interactions had distinct spatiotemporal kernels, with corresponding signatures in spontaneous activity. Excitability measured under optogenetic stimulus predicted spiking responses to subthreshold voltage, depolarization rate, and spike history during spontaneous firing. Genetic subtypes also showed different behavioral-state-dependent voltage-to-spike relationships despite similar population-average subthreshold voltages. These measurements link cellular properties and pairwise interactions to spontaneous circuit dynamics, establishing an approach for functional circuit mapping in vivo.

neuroscience↗

bioRxiv2026

Reformulating Protein Corona Prediction as Universal Protein-conditioned Learning

When nanoparticles enter biological environments, their surfaces are rapidly coated by proteins, forming a protein corona that governs their biological identity and downstream responses. Existing computational methods treat each protein as an independent prediction task, preventing the learning of shared adsorption principles and limiting generalization beyond experimentally characterized proteins. Here we introduce NanoPCT, a universal protein-conditioned learning framework that jointly models nanoparticle physicochemical properties and protein sequence representations. Using frozen ESM-2 embeddings and a joint interaction transformer, NanoPCT learns transferable representations of nano-bio interaction patterns across hundreds of proteins and maintains robust performance as protein diversity increases. The learned protein representation further enables zero-shot prediction for previously unseen proteins and biologically distinct protein groups through classifier reconstruction in embedding space. Beyond prediction, NanoPCT supports large-scale in silico inverse design and reveals consistent adsorption preferences between observed and unseen protein populations, suggesting that it captures transferable interaction patterns rather than protein-specific memorization. NanoPCT provides a unified framework for protein corona prediction, zero-shot generalization, and rational nanomaterial design.

bioinformatics↗

bioRxiv2026

Arsenic Impairs Critical Cellular Events of Human Cardiac Development Across Differentiation and Metabolic Maturation

Arsenic (As) is a widespread environmental contaminant, with exposure occurring primarily through contaminated food and water or cigarette smoke. Maternal exposure to As represents a significant risk factor for congenital heart disease in offspring. However, the mechanisms underlying As induced developmental cardiotoxicity remain poorly defined. Pluripotent stem cells provide an excellent platform for studying mechanisms of developmental toxicity. Embryoid bodies (EBs) derived from stem cells, can recapitulate many key events of early embryogenesis including germ layers specification. Human cardiomyocytes are derived from mesodermal progenitors. NKX2.5 governs cardiac specification and morphogenesis, and its mutation produces atrial and ventricular defects in humans. Using an NKX2.5 reporter stem cell line, we modeled mesoderm differentiation in EBs and cardiac induction in a 2D cardiomyocyte differentiation system. In an 8 day differentiation protocol, As (0.05 M; 3.75 ppb) significantly suppressed cardiomyocyte differentiation and cardiac induction. However, As increased mesodermal markers (HAND1, SNAI2, HOPX) gene expression. In contrast, cardiac specific genes, including NKX2.5, GATA4, Troponin T, and alpha actinin, were markedly downregulated in the presence of As. As also reduced expression of genes related to cardiac induction, fibroblast function, and vascular markers in the cardiac organoid model. As impaired cardiomyocyte maturation related gene expressions at concentrations without affecting cardiomyocyte proliferation. In a zebrafish model of early heart development, As increased heart rate and disrupted ion channel gene expression. In conclusion, As inhibits ESC differentiation into cardiomyocytes and disrupts cardiomyocyte maturation. Combining stem cell derived models with zebrafish exposure, this study illuminates adverse outcome pathways for As induced congenital heart defect.

pharmacology and toxicology↗

bioRxiv2026

Neutrophils orchestrate pre-metastatic immunosuppressive bone marrow remodeling during prostate cancer metastasis

The dynamic reprogramming of the bone marrow (BM) microenvironment during metastatic progression is poorly defined. Through single-cell profiling of BM samples spanning benign conditions, high-risk bone metastatic potential localized prostate cancer (HMP) and established bone metastatic prostate cancer (PCa), we uncovered a primary tumor-to-BM signaling axis that orchestrates niche formation through neutrophil-mediated remodeling. Tumor-associated neutrophils (TANs) emerge during the HMP stage, persist throughout metastatic progression, correlate with poor patient prognosis, and exhibit direct immunosuppressive activity that contributes to a BM environment permissive to metastatic colonization. Functional and genetic studies showed that tumor-associated systemic IGF1 reprogrammed BM neutrophils through IGF1R-MAPK/ERK-KLF4 signaling, with IL-1{beta}, reactive oxygen species and nitric oxide contributing to TAN-mediated T cell suppression. Tumor-cell Igf1 perturbation, IGF1R blockade and neutrophil-specific Igf1r deletion attenuated TAN-associated BM remodeling, and reduced subsequent bone metastatic progression. In bone-metastasis models, combined IGF1R and PD-1 blockade suppressed bone-metastatic progression. Translationally, our investigator-initiated clinical trial (NCT06866548) of teprotumumab plus tislelizumab in patients with established metastatic castration-resistant prostate cancer (mCRPC) provided initial safety data and exploratory clinical observations. Together, these findings identify IGF1R-dependent neutrophil reprogramming as a functionally important component of the evolving PCa bone metastatic microenvironment and provide a rationale for therapeutic targeting of IGF1R signaling together with immune checkpoint blockade.

cancer biology↗

bioRxiv2026

An integration-free CRISPR platform for genetic dissection of respiratory virus host dependencies in human nasal epithelium

Respiratory viruses, including coronaviruses, generally initiate infection in the upper respiratory tract, where mechanistic studies of host-virus interactions require models amenable to genetic manipulation. Human nasal air-liquid interface (ALI) cultures are realistic models of the nasal epithelium, but robust CRISPR/Cas9 engineering workflows for this primary cell system have been lacking. Here, we established an integration-free CRISPR/Cas9 workflow for genetic engineering of human nasal basal cells, which are subsequently differentiated into gene-edited ALI cultures. GFP-based sorting of RNP-transfected nasal basal cells substantially increases editing, achieving knockout efficiencies of 70-90% across gene targets (B2M, SLC35A1, ACE2, and TMPRSS2). Genetic disruption of ACE2 markedly reduces infection by SARS-CoV-2 and hCoV-NL63, confirming its important role for both viruses in the human nasal epithelium. TMPRSS2 knockout substantially reduces replication of both viruses, demonstrating SARS-CoV-2 depending on TMPRSS2 in the human nasal epithelium and providing genetic evidence in primary human cell cultures that hCoV-NL63 relies on TMPRSS2. This platform enables systematic genetic perturbation in a physiologically relevant model of the primary site of respiratory virus infection, validates ACE2 and TMPRSS2 as targets for medical countermeasures against SARS-CoV-2 and hCoV-NL63, and provides a versatile approach to identify host dependency factors for both circulating and emerging respiratory viruses.

microbiology↗

bioRxiv2026

Autonomic and neurotrophic signaling in alcohol-associated liver disease

Alcohol-associated liver disease (ALD) is a leading cause of liver-related morbidity and mortality, yet its pathogenesis remains incompletely understood. While neural regulation is critical for hepatic homeostasis, how autonomic and neurotrophic signaling is regulated in the liver during ALD progression remains largely unexplored. Here, we performed an integrative analysis of bulk and single-cell transcriptomic datasets of human livers across the spectrum of ALD. We identified stage-dependent remodeling and coordinated expression patterns of autonomic and neurotrophic genes. Autonomic signaling exhibited a consistent decline, with reduced expression of sympathetic (DBH, TH) and the parasympathetic (ChAT) markers. In contrast, pan-neuronal and neuroplasticity-associated genes (UCHL1, TUBB3, STMN2) were upregulated, indicating active neural remodeling. In parallel, factors such as MANF, GDNF, and NRG2, with the receptor NTRK1, were upregulated and positively correlated with clinical indicators of disease severity, inflammation and fibrosis. In contrast, trophic factors including NTF3 and CDNF, and receptors such as GFRA1 and GFRA3, were downregulated and inversely associated with these parameters. Single-cell analyses revealed that neurotrophic factors are predominantly expressed by hepatic stellate cells, supporting their role as resident neuroglia-like cells in the liver. In another independent cohort, we observed a coordinated downregulation of NTF3 and sympathetic nerve markers in alcohol-associated hepatitis (AH), suggesting impaired NTF3-mediated neurotrophic support may contribute to sympathetic dysfunction in AH. Collectively, these findings uncover previously unrecognized neuro-regulatory dimensions in ALD and suggest that dysregulated autonomic and neurotrophic signaling may contribute to disease progression. Conflict of interestThe authors have declared that no conflict of interest exists.

bioinformatics↗

bioRxiv2026

A long-term organotypic slice culture model of patient high-grade glioma preserving tumor heterogeneity and architecture, and specificity of CAR T cell targeting

ABSTRACT Background: High grade glioma (HGG), encompassing IDH-wildtype glioblastoma (CNS WHO grade 4), diffuse midline glioma (CNS WHO grade 4), oligodendroglioma (IDH-mutant and 1p/19q-codeleted) CNS WHO grade 3, together with IDH-mutant astrocytoma of CNS grade 3 and grade 4, represents the most lethal group of adult diffuse gliomas. Despite significant efforts, obtaining improvements in patient survival has been limited, in part, by difficulties in comprehensive ex vivo modeling of the complexities of HGG. Although organotypic slice cultures and other platforms have shown promise, existing models often fail to capture the intrinsic cellular heterogeneity, microenvironment, and architecture of HGG, restricting investigations into their fundamental biology as well as design of novel therapeutics. Methods: We developed a patient-derived HGG organotypic slice culture (GOSC) platform demonstrating long-term preservation of intrinsic tumor characteristics. Vibratome-sectioned slices were cultured in defined medium under conditions approximating those in situ: reduced oxygen (4%), low glucose (2.5 mM), and no added growth factors. The characteristics of GOSCs after 28 days in culture, including expression of immunotherapy targets, were compared to parental tumors using histological staining, immunofluorescence, multiplexed PhenoCycler (CODEX) imaging, bulk RNA-seq expression profiling, and CAR T cell killing assays. Results: GOSCs preserved the histopathology, cell heterogeneity, tumor microenvironment, and tissue architecture of their parental tumors throughout at least 28 days in culture. Importantly, GOSCs retained parental patterns of immunotherapy target expression, and CAR T cells co-cultured with GOSCs demonstrated target-specific killing consistent with parental tumors. Conclusions: This ex vivo GOSC platform offers long-term preservation of the cellular and structural properties of parental HGG tumors, thereby providing a robust tool for extended preclinical studies, and facilitating development and evaluation of advanced tumor-targeting strategies.

cancer biology↗

bioRxiv2026

Peptide frontier-orbital gaps narrow by backbone electrostatics

Aromatic side chains carry the optical and redox chemistry of peptides, and lengthening one is widely expected to extend its conjugation and narrow its HOMO-LUMO gap. We test that expectation with plane-wave density functional theory on 22 isolated neutral peptides NH2-(X)n-COOH (X = Phe, Tyr, Trp, Cys; n = 1 to 5, and n = 10 for Cys and Trp). The gap does narrow, at the same rate for all three aromatic residues, i.e., -0.17 to -0.18 eV per residue, even though benzene, phenol and indole are very different chromophores. Residue-resolved projections reveal why: nothing delocalises. Every aromatic frontier orbital is 97 to 100 % confined to a single residue and the HOMO participation number saturates at one ring. Each residue instead keeps its own local gap: for Trp it is invariant to within 0.03 eV over n = 2 to 10, while over that same span the global gap falls by 1.19 eV. The global gap is therefore an extreme-value functional of the residue site energies, gap(n) = -{Delta}(n), whose alignment penalty accounts for the entire effect: the polar backbone generates a monotonic internal electrostatic ramp that leaves the two gap edges at opposite ends of the chain, 32 [A] apart. We set out the resulting design rules for peptide-based bio-organic semiconductors.

biochemistry↗

bioRxiv2026

A Novel NO Donor 3A72 Activates Mitochondrial-Associated Endoplasmic Reticulum Membrane Structures to Induce Ferroptosis in Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) remains a major clinical challenge due to a lack of effective interventions and severe drug resistance. Disrupting tumor metabolism using nitric oxide (NO) donors to drive ferroptosis represents a promising therapeutic strategy. Here, we developed 3A72, a novel furan-coumarin-based NO donor with potent anti-TNBC efficacy. In vitro and in vivo studies demonstrated that 3A72 significantly suppresses TNBC cell proliferation and restricts xenograft tumor growth. Notably, its efficacy matches the first-line drug paclitaxel but with enhanced safety and reduced systemic toxicity. Mechanistically, 3A72 induces ferroptosis in triple negative breast cancer (TNBC) cells via a synergistic dual-pathway mechanism. Initially, 3A72 releases intracellular NO to trigger cytoplasmic calcium overload, followed by high-affinity binding to both inositol 1,4,5-trisphosphate receptor (IPR) and acyl-CoA synthetase long-chain family member 4 (ACSL4). Binding to IP3R provokes mitochondrial calcium overload and a massive burst of reactive oxygen species (ROS), while concurrent ACSL4 activation drives essential membrane phospholipid remodeling. Crucially, our transcriptomic and structural predictions reveal that while ACSL4 exhibits a broader distribution across organelle membranes, its functional association with IP3R spatially converges at the endoplasmic reticulum membranes associated with mitochondria (MAMs). This subcellular niche closely couples IPR-mediated ROS generation with ACSL4-driven lipid remodeling, accelerating lethal malondialdehyde (MDA) accumulation and triggering irreversible ferroptosis. Overall, these findings highlight the MAMs calcium-lipid metabolic hub as a promising structural scaffold for antitumor drug development.

cancer biology↗