Traditional drug development poses significant financial and temporal costs, whereas drug repurposing emerges as a cost-effective and efficient alternative. As large-scale biological networks proliferate, computational drug repurposing has become feasible, yet accurately capturing intricate heterogeneous network structures remains a persistent challenge. To address this challenge, we introduced a novel approach, called DRQuantum: Drug Repurposing via Quantum walks. Unlike random walks, quantum walks dispense with independence and harness quantum entanglement to simultaneously explore multiple paths, enabling faster traversal of networks. Moreover, DRQuantum accounts for both the local and global network structures. In this study, we constructed a heterogeneous multi-layer network by integrating drug-drug, disease-disease and protein-protein interaction networks. We then employed quantum walks to learn low-dimensional feature representations of nodes in these heterogeneous networks, ultimately inferring candidate drugs for repurposing beyond their original indications. Consequently, we observed that DRQuantum outperforms traditional drug repurposing methods in terms of AUROC, AUPRC and accuracy. Additionally, case studies for several specific diseases further validate the practical utility of our proposed method.
Feline infectious peritonitis (FIP), caused by feline coronavirus (FCoV) remains a critical therapeutic challenge with no universally approved treatments. We employed a drug repurposing strategy targeting the viral main protease (3CLpro), essential for FCoV replication. Virtual screening of FDA-approved drugs identified 15 candidates, with threesaquinavir (antiretroviral), lumacaftor (CFTR corrector), and gliquidone (antidiabetic)demonstrating notable antiviral activity. Postentry antiviral assays (cytopathic effect reduction, immunoperoxidase staining, RT-qPCR) yielded EC50 values of 0.25-0.48 μM (saquinavir), 30.32-58.49 μM (lumacaftor), and 48.85-51.28 μM (gliquidone). We validated 3CLpro inhibition using an intracellular dual-luciferase reporter assay designed to simultaneously account for compound cell penetration and metabolic stability. This confirmed specific protease inhibition with IC50 values of 23.70 ± 1.38 μM (saquinavir) and 38.68 ± 1.59 μM (lumacaftor), and 82.52 ± 1.50 μM (gliquidone). Synergy analysis with established anti-FIP agents (GC376, remdesivir, GS-441524, and molnupiravir) using SynergyFinder 3.0 revealed strong synergistic interactions: saquinavir + GC376 (mean ZIP score: 54.59), lumacaftor + GC376 (mean ZIP: 21.44), and gliquidone + remdesivir (mean ZIP: 24.13). Notably, optimal synergistic doses achieved 0.97-2.03 log10 viral suppression using only 20-50% of individual drug EC50 values, concurrently modulating viral-induced cytokine expression (TNF-α, IFN-β, and IL-6) in CRFK cells (p < 0.05). These rational combinations enable substantial dose reduction, offering practical strategies to improve treatment accessibility, reduce costs, and minimize adverse effects. This study establishes a framework for repurposing FDA-approved drugs in FIP therapy and supports translational evaluation of these combination regimens.
Mycobacterium abscessus is a rapidly growing nontuberculous mycobacterium characterized by extensive antibiotic resistance, presenting substantial challenges in clinical management. This study aimed to identify potential antimicrobial agents repurposed from an FDA-approved drug library to combat M. abscessus infection. An initial high-throughput screen of 2680 FDA-approved drugs was conducted to identify compounds with antimicrobial activity against M. abscessus. Promising hits were further assessed through resazurin-based minimum inhibitory concentration (MIC) assays and MTT cytotoxicity assays using multidrug-resistant (MDR)-M. abscessus clinical isolates and various human cells, respectively. Moreover, the interactions of hit compounds with conventional antibiotics were assessed using the checkerboard assay. The primary screen identified 39 nonantibiotic compounds with antibacterial activity against M. abscessus. Of these, eight exhibited favourable therapeutic indices on the basis of the MIC and cytotoxicity profiles. Among them, bismuth tripotassium dicitrate (BTD) emerged as the most promising candidate, demonstrating potent activity against multidrug-resistant clinical isolates, minimal cytotoxicity to multiple human cell lines, and significant synergy with clarithromycin and erythromycin. BTD has strong efficacy against M. abscessus and favourable safety and synergy profiles. These findings support its potential for repurposing as a novel therapeutic for M. abscessus infections.
Indwelling urethral catheters are the most widely used medical devices across the world, and catheter-associated urinary tract infections (CAUTIs) are the most common type of healthcare acquired infection. For many patients, urinary catheter blockage is a common and recurring problem, which can have considerable negative impact on patient health and well-being. Blockage primarily stems from the formation of crystalline bacterial biofilms on catheter surfaces, which can lead to upper urinary tract infection (UTI) and the onset of serious clinical complications. Potential solutions to this important clinical problem include the development of novel antibiofilm agents that can prevent formation of these communities on urinary catheters. However, traditional de novo methods of drug discovery are laborious, expensive, have long lead times and carry a high risk of failure in the clinical trial stages. One potential approach to mitigate this risk and cost, is the evaluation of pre-existing licensed drugs for those with useful antibiofilm or antimicrobial activity. Here we review current preclinical evidence for antibiofilm and antimicrobial activities in licensed drugs from a range of classes, such as urease inhibitors, Selective Serotonin Reuptake Inhibitors (SSRIs), phenothiazines, oncology therapeutics and non-steroidal anti-inflammatory drugs (NSAIDs). In doing so, we consider the application of the repurposing approach to control CAUTI and catheter blockage, and identify key challenges and opportunities related to delivery of repurposed drugs to the catheterised urinary tract.
Statins are marketed as antihyperlipidemic drugs, have been repurposed in cancer management. Their repurposing is based on antiproliferative, apoptotic, and anti-metastatic properties. Head and neck cancer, including oral carcinoma, remains a global burden with limited advancements. Therefore, we study the preclinical and clinical advancements of statins and statins-loaded nanocarriers in head and neck cancer. Data were extracted from various sources, including PubMed, Scopus, and Web of Science. Publications reporting in vitro, in vivo, and clinical data were evaluated. Data will be extracted and assessed following PRISMA guidelines. Various statistical parameters, such as Risk Ratio and Odds Ratio, were computed to analyze the reported clinical trials. This systematic review provides evidence-based proof of the anticancer potential of statins. Research gaps and future perspectives have also been discussed.
To date, there is no proven licensed systemic treatment for neurofibromatosis type 2 (NF2)-related schwannomatosis patients. There is a need for more effective, less toxic treatments and, as a rare disease, NF2 is often overlooked in targeted drug development. Subcutaneous schwannomas of the skin (CS) are common in the NF2 population.This trial involves the repurposing of medications already licensed for HIV-ritonavir and lopinavir (Kaletra and Norvir)-that have been shown to reduce tumour growth by reducing cell proliferation in human schwannoma and meningioma tumour cell cultures. The safety and tolerability of these drugs are already known, so they are safe candidates to trial in NF2 patients. This trial is an open-label, phase 0 design. A maximum of 16 participants diagnosed with NF2-related schwannomatosis will be enrolled in this study. Treatment duration is 30 days, with a 30-day follow-up. Biopsies and blood samples will be collected to assess whether the drugs reach the tumours and to analyse the tumour-cell response. The primary outcome is pharmacodynamic response, defined as a statistically significant decrease in biomarker activity in CS biopsy samples at day 30. The sample size calculation is based on the tissue biomarker response. The study was approved by an Ethics Committee (West of Scotland Research Ethics Service (23/WS/0178)), the Health Research Authority (HRA), the Medicines and Healthcare products Regulatory Authority (MHRA) and each of the participating NHS Trust's Research and Development departments. Following analysis of trial data, the trial results will be written up for publication in a peer-reviewed scientific journal and will be disseminated at conferences. ISRCTN10422213.
Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder with significant involvement of neuroinflammation, for which the current interventions are limited in efficacy. Certain antihistamines such as clemastine have neuroprotective properties beyond H1 receptor antagonism, including anti-inflammatory, antioxidant, remyelinating effects, modulates neuroinflammatory pathways. This study aims to investigate the neuroprotective potential of clemastine via in silico and in vivo studies for repurposing against AD. In silico analyses involved network pharmacology, molecular docking, and 100-ns molecular dynamic simulations along with principal component analysis and binding free energy calculations. Whereas, experimental studies involved treatment of clemastine (5 mg/kg and 10 mg/Kg, p.o.) for 14 days in lipopolysaccharide-induced neuroinflammatory (250 µg/Kg, i.p.) rat. Behavioral assessment was performed using Morris water maze (MWM) test. Biochemical parameters including acetylcholinesterase (AChE) activity, oxidative stress markers (MDA, SOD, CAT, GSH), and inflammatory biomarkers (NLRP3, TNF-α, IL-1β) were evaluated. Histopathological analysis of hippocampal CA3 region was performed using Nissl's staining. Network analysis identified 52 overlapped targets between clemastine and AD. Hub genes such as GSK3β, DRD1, DRD2, CHRNA4, and SLC6A4 were associated with neurotransmission and kinase signaling pathways. Enrichment analysis highlighted PI3K/Akt, MAPK, and neuroactive ligand-receptor interaction pathways. Molecular docking and molecular dynamic simulations confirmed stable binding of clemastine with GSK-3β, PI3K, and NLRP3 proteins. Animal model studies demonstrated that clemastine significantly improved cognitive performance in MWM (p < 0.001), reduced AChE level (p < 0.0001), restored antioxidant enzyme levels, suppressed inflammatory mediators (p < 0.0001), and preserved hippocampal neuronal structure. The study provides novel integrative evidence linking its antihistaminic action with simultaneous regulation of neuroinflammation through multi-target modulation of inflammation, oxidative stress, and neuronal signaling pathways, highlighting its potential as a promising repurposed therapeutic candidate for AD.
The microbiome-gut-brain axis (MGBA) redefines the field of anti-inflammatory therapeutics by shifting from targeting isolated immune pathways in an organ to a bidirectional, integral network, composed of gut microbiota, the peripheral immune system, barrier tissues, and neural circuits. Such reframing is predominantly pertinent for certain CNS and systemic diseases, including Alzheimer's disease and ischemic stroke. These diseases have been resistant to conventional anti-inflammatory agents that either poorly pass the blood-brain barrier or do not inhibit the upstream inflammatory drivers. Increasing evidence demonstrates that microbial communities and their metabolites modulate systemic and central immune tone, and affect barrier integrity, hence brain inflammation frequently presents with a measurable fingerprint in the gut and vice versa. Targeted microbiome therapies, metabolite replacement, and biomarker-guided patient stratification all emerge as promising new strategies when acknowledging the gut is a modifiable source. Mechanistically, the crosstalk between neuro-immune-microbial trails creates concrete therapeutic entry points. These include: (1) altering microbial enzymes that produce pro-inflammatory metabolites, (2) delivering beneficial metabolites to support homeostatic glial states, and (3) modulating afferent neural pathways like vagus nerve stimulation, which triggers the cholinergic anti-inflammatory pathway (CAP). The future of MGBA-targeted therapeutics will mostly repurpose current anti-inflammatory agents and neuromodulatory strategies into combinatorial ones that pair source control (gut-restricted drugs) with targeted central modulation (microglial modulators such as minocycline) and neural orchestration (vagus nerve stimulation, VNS). These repurposing strategies are increasingly guided by baseline inflammatory and microbiome biomarkers and may help in establishing new therapeutic paradigms for these intersecting disorders of the gut, immune system, and brain in appropriately defined patient subgroups.
Fenofibrate, a well-established peroxisome proliferator-activated receptor alpha (PPARα) agonist, is widely used clinically for managing hypercholesterolemia and hyperlipidemia. In recent years, its potential anti-tumor properties have garnered considerable attention. Accumulating evidence demonstrates that fenofibrate inhibits the growth of various malignancies, including breast, liver, prostate, and lung cancers, by modulating lipid metabolism, suppressing cancer cell proliferation, and inducing apoptosis. The anti-tumor mechanisms of fenofibrate involve both PPARα-dependent pathways through direct regulation of PPARα and associated lipid metabolic enzymes, and PPARα-independent pathways, such as inhibition of the Akt/mTOR, TNF-α/NF-κB, and Cx43/EGF/ERK1/2 signaling axes. Furthermore, fenofibrate exhibits synergistic effects with conventional chemotherapeutic agents (e.g., paclitaxel and docetaxel) by enhancing T-cell viability, activating immune responses, and thereby improving chemotherapy sensitivity. This review comprehensively summarizes the efficacy and underlying mechanisms of fenofibrate against diverse cancer types and discusses the potential applications and challenges associated with its repurposing for cancer therapy. A deeper understanding of fenofibrate's anti-cancer capabilities and molecular targets may provide novel insights for the development of innovative therapeutic strategies.
Earlier detection is strongly associated with increased survival for women with ovarian cancer. Unfortunately, current screening strategies, employing serial serum or ultrasound assessments, lack adequate sensitivity and specificity for use in the low-prevalence general population. In contrast, screening for cervical cancer by Pap tests has been routinely performed for over 50 years. Since ovarian cancer cells have been observed in Pap tests, ovarian cancer protein biomarkers may also be present; yet Pap samples have not been rigorously examined for diagnostic proteins. Assessment of cervical effluent, as can be collected in a Pap test, has the potential to differentiate ovarian cancer cases from healthy controls, and thereby demonstrate that intra-abdominal pathology may be detected using this commonly acquired specimen. We hypothesize that proteins shed by ovarian cancer cells can be detected in the SurePath™ liquid-based Pap test fixative using mass spectrometry (MS)-based proteomics, making it possible to distinguish women with ovarian cancer from healthy women. Candidate ovarian cancer biomarkers were successfully identified in liquid-based Pap test samples from 20 cases of high grade serous ovarian cancer, 10 benign ovarian conditions, and 10 healthy control samples, by performing Tandem Mass Tag™ isobaric labeling, 2D liquid chromatography-MS/MS, and bioinformatics integration. Selected reaction monitoring (SRM) MS-based targeted proteomics was then performed using a panel of candidate biomarkers to quantify their abundance in an expanded patient cohort of 90 liquid-based Pap tests. A multi-protein classifier was developed using the SRM-MS data comprised of 6 proteins and achieving an AUC of 0.880 (95% CI: 0.738-0.989); sensitivity at 90% specificity was 0.800. This pilot study provides evidence that the cervical effluent collected in SurePath™ fixative from Pap tests has the potential to be used as a biospecimen for the detection of ovarian cancer proteins. Our long-term goal is to develop a noninvasive screening test that can be incorporated into a routine Pap test or a self-administered home test, so that women can be screened simultaneously for cervical and ovarian cancer.
Integrated computational-experimental platforms are promising for GPCR drug discovery, but systematically connecting high-resolution structural prediction with functional validation remains challenging. Here, we apply an integrated platform to discover a novel, potent antagonist targeting the P2Y14 receptor (P2Y14R), a key target in inflammatory diseases. Combining structure-based virtual screening, BPMD, all-atom molecular dynamics simulations, and cellular cAMP functional assays, our platform pinpointed Benfotiamine, an approved drug, as a nanomolar-potency P2Y14R antagonist (IC50 = 0.31 nM). Computational analyses indicated that Benfotiamine exerts a "conformational lock" to stabilize the inactive receptor and hinder Gi coupling. Subsequent CETSA and SPR experiments confirmed its direct binding to the target. Functionally, Benfotiamine exhibited therapeutic efficacy in murine models of dextran sulfate sodium-induced colitis and monosodium urate-induced gouty arthritis. Beyond presenting a promising repurposed anti-inflammatory agent, this study validates a modular and extensible computational-experimental integration platform that can be widely applied to expedite the discovery and mechanistic characterization of GPCR-targeted therapeutics.
Population cancer screening detects the presence of early-stage disease rather than assessing future disease risk. We evaluated whether widely implemented cardiovascular disease (CVD) risk models can predict 10-year cancer risk, comparing them with the QCancer risk model. We evaluated four CVD prediction models: QRISK3, the Pooled Cohort Equations (PCE), SCORE2 and SCORE2-OP. The models were recalibrated using 20% of the UK Biobank (UKB) cohort and tested in the remainder and in the Clinical Practice Research Datalink (CPRD). We gauged model performance by discrimination (c-statistics) and calibration (slope and intercept) and evaluated feature importance. In the UKB test set, the c-statistics for incident CVD ranged from 0.71 to 0.74 (9,712 events). All CVD models achieved a c-statistic of 0.63 for any cancer (23,010 events) and showed CVD-equivalent discrimination for gastro-oesophageal, liver and biliary tree, laryngeal, renal tract, and lung cancers (c-statistic range: 0.70;0.81). The recalibrated CVD models showed near-perfect calibration (median intercept 0.01, Q1;Q3 -0.05;0.03 and slope 1.01, Q1;Q3 0.95;1.14). Performance in CPRD (393,622 cancer events) was similar: the median c-statistic, calibration intercept, and slope were 0.01 (95%CI 0.00;0.02), 0.09 (95%CI 0.03;0.20), and 0.04 (95%CI 0.01;0.14) higher, respectively, in CPRD than in UKB. After age, smoking status and systolic blood pressure were the most influential predictors of cancer risk. Widely implemented CVD prediction models perform similarly to the less widely used QCancer models in the prediction of incident cancers. They may be used to inform cancer prevention and guide risk-stratified monitoring. The recalibrated models are available through an open source web application.
In this study, we explicitly evaluate the anti-inflammatory effects of ivacaftor as a route-of-delivery comparison (intraperitoneal vs. intratracheal) in a reproducible LPS-induced lung inflammation model. We evaluated the effects of ivacaftor [40 mg/kg, administered either intraperitoneally (IP) or intratracheally (IT)] on lipopolysaccharide (LPS, 8 μg)-induced lung inflammation in 8-10-week-old female C57BL/6 mice. Mice were euthanised at 24 or 72 h post-treatment, and serum, bronchoalveolar lavage fluid (BALF), and lung tissue were collected for analysis. Inflammation was assessed by measuring immune cell infiltration, fluorescence-activated cell sorting (FACS)-based leucocyte profiling, and histological changes. Ivacaftor concentrations in the samples were quantified using multiple reaction monitoring liquid chromatography-mass spectrometry (MRM-LCMS). At 24 h, both IT and IP ivacaftor significantly reduced the total BALF cell counts (∼40% reduction) and neutrophil infiltration compared to the LPS-only controls (p < 0.05). Inflammatory responses were reduced following ivacaftor treatment, as reflected by lower BALF cellularity, reduced neutrophil infiltration, and improved histological outcomes. Histological analysis showed reduced alveolar wall thickening and immune cell infiltration in ivacaftor-treated lungs. FACS confirmed lower frequencies of neutrophils and macrophages. Ivacaftor concentrations were higher in lung tissue following IT administration. Our findings indicate that ivacaftor has a potential anti-inflammatory effect, unrelated to cystic fibrosis transmembrane conductance regulator (CFTR) function, in a murine model of LPS-induced acute lung injury. These results offer insights into inflammation mechanisms and could inform future clinical treatments.
Helicobacter pylori colonizes the gastric mucosa of around half of the world's population and is a major cause of chronic gastritis, peptic ulcer disease, and gastric cancer. Current therapies are becoming increasingly ineffective due to the rapid spread of antibiotic resistance, creating an urgent need for new treatment options with distinct mechanisms of action. Drug repurposing offers a practical and cost-effective approach to address this gap. PBT2 is an 8-hydroxyquinoline derivative originally developed for the treatment of neurodegenerative diseases and has more recently been shown to possess antimicrobial activity. In this study, we demonstrate that PBT2 displays potent bactericidal activity against H. pylori, including multidrug-resistant clinical isolates. PBT2 rapidly killed H. pylori in vitro at low concentrations, with faster killing kinetics than commonly used antibiotics, and no resistance was detected after 30 days of continuous exposure. Importantly, PBT2 was effective in clearing an H. pylori infection in a murine model. Quantitative sequential window acquisition of all theoretical-mass spectrometry proteomic analysis revealed that PBT2 triggers broad disruption of essential bacterial processes, including global suppression of translation, impairment of iron-sulfur cluster assembly and respiration, dysregulation of metal homeostasis, and reduced abundance of virulence- and motility-associated proteins. We reported that PBT2 can act as a nickel ionophore, with Ni2+ being the highest-affinity ligand for PBT2 reported to date. Together, these findings suggest that PBT2 acts through a multifaceted, metal-dependent mode of action that limits the potential for emergence of resistance. Our work highlights PBT2 as a promising candidate for repurposing to treat multidrug-resistant H. pylori infections.IMPORTANCEAntibiotic resistance is steadily reducing our ability to treat common bacterial infections, while the development of new antibiotics has slowed. Helicobacter pylori is a clear example of this growing problem, with treatment failures becoming more common worldwide. This study highlights the value of taking a different approach by repurposing existing drugs for new antibacterial uses. Rather than acting on a single bacterial target, the compound examined here disrupts multiple essential processes at once, reducing the probability of resistance developing.
<SPAN style="font-weight: 400;">The QueC protein family (PF06508) is best known for its role in the biosynthesis of 7‑deazaguanine derivatives, including the tRNA modification queuosine and the DNA base 7‑cyano-7‑deazaguanine (preQ</SPAN><SPAN style="font-weight: 400;">0</SPAN><SPAN style="font-weight: 400;">). Recent discoveries, however, reveal that this ancient scaffold has been repeatedly repurposed for distinct biological functions, including roles in anti‑phage defense. Here, we combine Sequence Similarity Networks (SSN) with genomic neighborhood analyses to map the functional diversification of the PF06508 superfamily. We delineate the canonical QueC cluster and experimentally define a refined catalytic signature, identifying previously unrecognized residues essential for tRNA modification. Beyond the canonical function, we characterize the evolutionary repurposing of the QueC fold in anti-phage systems, distinguishing between the "divergent specialist" QatC, which remodels the active site, and the "minimalist conservationist" Cap9, which retains the ancestral catalytic core. We further uncover the expansion of the superfamily into additional biochemical pathways, including cofactor biosynthesis (e.g., LarE) and purine salvage. Together, these findings provide a comprehensive framework for understanding how the QueC scaffold has been adapted from small‑molecule biosynthesis to roles in protein modification and phage defense.</SPAN>.
Drug resistance limits the efficacy of anthracycline-based chemotherapy in breast cancer. This study investigated whether atorvastatin (ATR) enhances doxorubicin (DOX) activity by coordinating the disruption of multiple resistance pathways in MCF-7 cells. Cytotoxicity and drug interaction were quantified by MTT assay and Chou-Talalay analysis, respectively. Modulation of the cell death processes was assessed at the molecular level by evaluating Bcl-2 and Bax expression with qRT-PCR, while at the protein level, apoptosis was analyzed by Annexin V assay using flow cytometry. Global DNA methylation patterns were determined using 5-methylcytosine (5-mC)-targeted ELISA. Molecular docking simulations characterized ATR's binding interactions with P-glycoprotein (P-gp) and Heat Shock Factor-1 (HSF-1). ATR and DOX produced consistent synergism (CI < 1) with a favorable dose-reduction index (DRI > 1) for DOX. The ATR-DOX combination also markedly shifted the Bcl-2/Bax balance toward apoptosis and increased apoptotic cells from 13.5% to 62% (~4.6-fold). The combination suppressed DOX-induced upregulation of HSP90 and HSP60 and significantly mitigated global DNA hypermethylation. In silico docking predicted favorable binding of ATR to P-glycoprotein (-10.24 kcal/mol) and Heat Shock Factor-1 DNA-binding domain (-6.69 kcal/mol), supporting potential modulation of drug efflux and stress-response pathways. Collectively, atorvastatin enhances DOX cytotoxicity through integrated effects on efflux regulation, proteostasis, epigenetic regulation, and apoptotic priming. Given its established clinical safety, atorvastatin represents a practical candidate for repurposing to improve anthracycline-based breast cancer therapy.
Despite advances in treating atherosclerotic coronary artery disease (ACAD), the risk of major adverse cardiovascular events remains high. We aimed to identify potential therapeutic targets for ACAD by integrating genome-wide association studies (GWAS) and Mendelian randomization (MR) analyses. Leveraging druggable genome data, cis-eQTL/cis-pQTL from blood and coronary artery tissue, alongside GWAS summary data for coronary atherosclerosis, we performed drug-target MR to identify causal genes for ACAD at both transcript and protein levels. Phenome-wide association (PheWAS), protein-protein interaction (PPI) networks, enrichment analysis, molecular docking, and in vivo experiments were also conducted. MR identified 57 blood cis-eQTL, 50 blood cis-pQTL, and 5 coronary artery cis-eQTL druggable genes potentially causal for coronary atherosclerosis. Notably, CES2 and TGFA showed consistent protective associations in both cis-eQTL and cis-pQTL analyses. Genetically elevated blood CES2 and TGFA levels were significantly associated with reduced ACAD risk (p < 0.001). PheWAS revealed no significant adverse effects for TGFA targeting. PPI and enrichment analyses highlighted metabolic pathways. Ten drugs targeting CES2 and three targeting TGFA were identified; in vivo experiments confirmed that atorvastatin, simvastatin, prasugrel, and vicagrel upregulate circulating CES2, while cetuximab and panitumumab increase circulating TGFA. Genetically elevated circulating CES2 and TGFA levels are causally associated with reduced ACAD risk. Several candidate drugs capable of modulating these proteins might offer repurposing opportunities for ACAD prevention, warranting further mechanistic and clinical validation.
The initial COVID-19 outbreak precipitated unprecedented health crises, with research impeded by containment protocols and the high mortality rate. While SARS-CoV-2 perturbs immune ubiquitin dynamics, the causal interplay between them remains uncharacterized. We integrated multi-omics and Mendelian randomization (MR) approaches, including bidirectional and two-step MR, to assess causality between ubiquitination and initial COVID-19 severity and to determine whether immune cells mediate this relationship. Transcriptomic profiling, protein-protein interaction networks, immune-correlation analysis, and an elastic-net-derived prognostic model were employed to explore these interactions further and identify potential biomarkers. Bidirectional MR confirmed causal links between COVID-19 severity and ubiquitination, identifying 13 ubiquitin-related genes with altered expression. A two-step MR analysis of 731 immune cell traits revealed CD45RA+ CD8+ T cells as a mediator of severe COVID-19-induced ubiquitin dysregulation (mediation proportion: 5.797%). A prognostic model based on ISG15, UBE2B, and UBE2V1 demonstrated high predictive accuracy in the training cohort (AUC up to 0.977) and maintained good predictive power in the testing cohort (AUC: 0.672-0.819), with significant risk stratification (log-rank P < 0.001). We propose that ubiquitin may function as a causal immune orchestrator, shaping clinical outcomes in the initial phase of COVID-19. The derived ubiquitin gene prognostic model demonstrates potential for clinical risk stratification, while pathway enrichment and druginteraction analyses suggest opportunities for repurposing ubiquitin-related targets in COVID-19 therapy. This ubiquitin-centric framework advances precision management of severe COVID- 19, highlights the potential of ubiquitin-related targets for drug development, and establishes a multi-omics MR-based paradigm for future viral pandemics.
Corynebacterium glutamicum is a crucial food-grade (GRAS) bacterial chassis widely utilized for the industrial production of amino acids and nutraceuticals. However, the efficient production of recombinant proteins and secondary metabolites in this host remains limited by context dependence and low translational efficiency. To overcome this, we introduce a 5'-end translationalization strategy. By repurposing passive 5' untranslated regions (5'UTRs) into actively translated fore-cistrons, we converted conventional monocistronic designs into context-independent, leaderless polycistronic designs (PCDs). This assembly of concatenated fore-cistrons functions as a translational amplifier, largely decoupling protein output from mRNA abundance. We validated this platform by optimizing two biomanufacturing paradigms: achieving a 4.07-fold enhanced secretion of OmlA, a porcine vaccine antigen, and boosting biosynthesis of the food-grade pigment indigoidine to 1.20 g/L (a 7.33-fold increase over baselines). Together, this framework establishes a versatile, portable toolkit to overcome translational bottlenecks, enabling robust hyperproduction of recombinant proteins and engineered metabolites in biotechnology.
Major depressive disorder (MDD) remains difficult to treat with agents that are simultaneously rapid, durable, and well-tolerated. Propofol shows fast antidepressant-like actions but is limited by anesthetic liability, a short half-life, and poor brain-selective exposure. Here, we report Nap-FFYK(propofol), a peptide-propofol self-assembling prodrug engineered to decouple anesthetic burden from sustained antidepressant benefit. Physicochemical characterization confirmed the stimulus-triggered conversion of Nap-FFYK(propofol) to Nap-FFYK and propofol. The prodrug displays high drug loading, robust self-assembly into nanospheres, and carboxylesterase (CES)-responsive release, enabling on-site liberation of propofol with a lower risk of respiratory depression. In the chronic social defeat stress mouse model, a single administration produced rapid and durable antidepressant-like effects with a swift recovery from transient sedation. Mechanistic analyses demonstrated reduced microglial activation, restored astrocytic support, preserved blood-brain barrier integrity, and enhanced synaptic plasticity in hippocampal circuits. These findings validate a targeting-assembly release strategy for propofol repurposing and establish peptide self-assembly prodrugs as a tractable platform for brain-directed, rapid-acting antidepressant therapy. This approach provides a blueprint to pair hour-scale symptom relief with improved safety and durability, addressing key limitations of existing rapid-acting interventions.