Objective To elucidate the pharmacological mechanisms of worenine in pulmonary fibrosis (PF) through an integrative strategy combining network pharmacology, molecular docking, and experimental validation. Methods Potential targets of worenine and PF-related genes were obtained from public databases and integrated to construct interaction networks. Protein-protein interaction (PPI), GO, and KEGG enrichment analyses were performed to identify key targets and pathways. Molecular docking evaluated binding affinities between worenine and hub targets. The anti-fibrotic effects and mechanisms of worenine were validated in bleomycin-induced PF mice and in TGF-β1-stimulated A549 and MRC-5 cells. Results A total of 116 overlapping targets were identified. Key targets included SRC, IL6, TNF, NFKB1, and PIK3CA. Enrichment analyses indicated that worenine regulates pathways related to inflammation and cellular stress, including TNF, IL-17, and HIF-1 signaling. Molecular docking showed strong binding affinities between worenine and core targets such as EZH2, PTGS2, PBK, SRC, NFKB1, and IL6. In vivo, worenine alleviated PF, reducing collagen deposition and improving histopathology. In vitro, worenine inhibited EMT and FMT, accompanied by suppression of PBK/SRC-associated TGF-β1/Smad, ERK, and NF-κB signaling, as well as decreased secretion of IL-6, IL-8, and VEGFA. Conclusion Worenine attenuates experimental pulmonary fibrosis, potentially through modulation of PBK/SRC-associated signaling and the TGF-β1/Smad, ERK, and NF-κB pathways. These findings provide a rationale for further investigation of Worenine as a potential therapeutic candidate for PF.
Glioblastoma multiforme (GBM) exhibits strong resistance to radiotherapy, partly driven by glioblastoma stem-like cells (GSCs) with enhanced redox homeostasis and DNA repair capacity. This study evaluated whether targeting Nrf2-mediated antioxidant signaling and PARP1-dependent DNA repair enhances GSC radiosensitivity to different radiation modalities. Pharmacological inhibition of Nrf2 (ML385, 6 µmol/L) or PARP1 (olaparib, 5 µmol/L) reduced tumorsphere formation to 74.5 ± 10% and 58.56 ± 14.5% of control levels, respectively, while combined treatment further reduced formation to 51 ± 11% and sphere size to 29% of control. Western blotting confirmed effective pathway inhibition, with complete suppression of PARP activity and approximately 30% reduction in Nrf2 downstream proteins (SOD1, PRDX2, and NQO1). Dose-response analysis showed D₅₀ values of 5.03 ± 0.09 Gy (photons), 2.96 ± 0.91 Gy (protons), and 2.04 ± 0.47 Gy (carbon ions), corresponding to RBE₅₀ values of 1, 1.70 ± 0.55, and 2.46 ± 0.57, respectively. ML385 enhanced radiosensitivity to photons and protons and showed a similar radiosensitizing trend following carbon-ion irradiation, whereas olaparib showed its strongest effect with photons and limited effects with protons and carbon ions. Combined treatment produced a greater reduction in radiation survival than either inhibitor alone under selected conditions, particularly following photon irradiation. Nrf2 inhibition reduced downstream antioxidant proteins and increased late apoptotic/necrotic fraction, while PARP1 inhibition was associated with altered DNA damage persistence. Combined inhibition further increased γ-H2AX foci at selected time points following proton irradiation, consistent with delayed or incomplete repair of radiation-induced DNA damage. These findings support Nrf2 and PARP1 as potential regulators of GSC radioresistance and provide a rationale for further investigation of their therapeutic targeting in combination with radiotherapy.
Effective wound healing is often hindered by hypoxia, inflammation, and impaired angiogenesis. Although mesenchymal stem cell (MSC)-based therapies have shown potential, their clinical application remains limited. As a result, MSC-derived exosomes, particularly adipose-derived stem cell exosomes (ADSC-Exos), have been proposed as a cell-free alternative; however, they still face limitations, including suboptimal efficiency in targeted delivery. To address these critical obstacles in existing regenerative therapies, we isolated ADSC-Exos from adipose tissue and fabricated a novel ultrasound-responsive, carbomer-based hydrogel incorporating oxygen-enriched ADSC-Exo nanobubbles (Gel-Exo-NB+US). The preclinical wound-healing potential of the developed nanohydrogel was evaluated using a full-thickness wound model in Wistar rats. The engineered hydrogel exhibited favorable physicochemical properties, including shear-thinning and thixotropic behavior, enabling efficient topical application and enhanced tissue adhesion. In a rat full-thickness skin defect model, the combined Gel-Exo-NB and Gel-Exo-NB + US treatments achieved the highest wound-closure rate among all groups, with enhanced re-epithelialization, a reduced scar index, elevated CD31 and α-SMA expression, increased fibroblast proliferation, and increased blood vessel formation. Furthermore, unprecedented hair-follicle regeneration and abundant collagen deposition were observed in the Gel-Exo-NB + US-treated group compared with the Gel-Exo-NB-treated group, highlighting the pivotal role of ultrasound in skin defect regeneration. Notably, the resulting collagen deposition, abundant hair follicles, and thin epidermis suggest potential for scarless wound healing. These therapeutic outcomes are attributed to the synergistic effects of simultaneous ultrasound-mediated exosome therapy and oxygen delivery.
The gastrin-releasing peptide receptor (GRPR) is highly expressed in prostate and breast cancers and hence serves as a legitimate target for radionuclide therapy (TRT). Radiolabeled GRPR antagonists are attractive radionuclide-carriers for TRT, due to their rapid tissue penetration, fast blood/whole-body clearance and inherent biosafety. We hereby explore the impact of dosing frequency of a GRPR-radioantagonist with improved tumor retention on therapeutic efficacy. For this purpose, we first studied the biodistribution of the recently developed [177Lu]Lu-AU-SAR-M1 in mice to estimate dosimetry. Next, mice with PC-3 xenografts (GRPR-positive) were treated with 6 injections of [177Lu]Lu-AU-SAR-M1 (12 MBq/injection), administered weekly (Group 1) or in 2 cycles of 3 injections/week with a 4-week interval (Group 2); an additional Control group got vehicle injections. Body weight, tumor volume, and blood cell counts were monitored. No evidence of bone marrow toxicity was observed. Both treatment regimens significantly slowed down tumor growth and improved survival. Median survival in the Control group was 37 d, while 50% of tumors were eradicated in the treated groups (no significant difference between groups). These preclinical findings qualify [177Lu]Lu-AU-SAR-M1, as a promising candidate for TRT in patients. Results of this study might be used in the design of follow-up studies.
Background Remarkable success of anti-CD20 B cell depleting therapies in multiple sclerosis (MS) highlights antibody-independent role of B cells in driving inflammatory relapsing-remitting stage of the disease. Yet, long-term B cell depletion leads to increased infections, pointing to the need for more targeted treatments. Objectives To establish a high-throughput translational assay of primary B cells for drug screening. Results We present an assay of primary B cells isolated from MS patients and healthy individuals. We tested various methods of B cell isolation, compared impact of combinations of different stimuli, including 2-signal and 3-signal mode of activation at multiple seeding densities on production of proinflammatory cytokines Interleukin 6 (IL-6), Tumor Necrosis Factor (TNF) and Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF), using 384-well microplate with semi-automation. Ibrutinib, a small molecule that suppresses B cell proliferation and survival by inhibiting protein kinases, including Bruton's tyrosine kinase (BTK), was used to validate the miniaturized B cell assay for drug screening. Seven BTK inhibitors, Ibrutinib and six novel compounds currently under later stage clinical development for autoimmune disease, were evaluated with concentration-response as proof-of-concept validation of the assay. Conclusions An optimized primary B cells assay in 384-well microplate format was established and validated by Ibrutinib. Moreover, we demonstrated that six clinical stage BTK inhibitors considerably impact B cell activation and secretion of IL-6, TNF and GM-CSF.
Tumor-associated vascular dysfunction is a major barrier limiting the efficacy of cancer therapies by impairing drug delivery, restricting immune-cell infiltration, and promoting hypoxia within the tumor microenvironment (TME). While anti-angiogenic therapies aim to suppress abnormal vessel formation, excessive vascular inhibition may further worsen hypoxia and therapeutic resistance. Here, we investigated whether restoration of endothelial integrity using Poloxamer 188 (P188), a membrane-stabilizing triblock copolymer, could improve vascular function and enhance therapeutic efficacy in triple-negative breast cancer (TNBC). In vitro, P188 prevented and reversed endothelial leakiness, restoring endothelial barrier integrity and enhancing tight junction formation, as demonstrated by increased Zonula occludens-1 (ZO-1) expression, a tight-junction protein, and sealed endothelial morphology. In vivo, fluorescently labeled P188 accumulated in TNBC tumors and significantly enhanced Dextran delivery, indicating improved vascular function and tumor perfusion. Although P188 showed no direct cytotoxic effect on TNBC cells and did not enhance Doxorubicin activity in vitro, combination treatment significantly suppressed tumor progression in vivo, while simultaneously reducing treatment-associated weight loss. Importantly, P188 also enhanced antitumor immunity. While P188 did not directly activate T cells or increase their cytotoxicity in vitro, it significantly increased T-cell infiltration into tumors in vivo and enhanced the therapeutic efficacy of adoptively transferred human normal peripheral blood mononuclear cell (PBMCs). Collectively, these findings demonstrate that restoration of tumor vascular integrity using P188 improves both chemotherapy and immunotherapy efficacy by enhancing therapeutic delivery and immune-cell infiltration. This study highlights vascular restoration as a promising therapeutic strategy for overcoming TME-mediated resistance in solid tumors.
β-thalassemia is a group of hereditary, autosomal recessive blood disorders caused by reduced synthesis or absence of β-globin chains. Reactivation of hemoglobin F (Hb-F) by pharmacological means is an excellent remedial approach for treating β-thalassemia. Hydroxyurea is currently the only approved drug for Hb-F induction therapy. However, the variable efficacy and high cytotoxicity concerns of hydroxyurea have limited its utilization in clinical practices. This preclinical investigation demonstrated the potential of the antiviral drug entecavir (ETV) as an Hb-F inducer using the K562 cell line and validated in a β-YAC transgenic mice model. In the K562 cell line and β-YAC transgenic mice, the effects of ETV on Hb-F induction were investigated in a dose dependent manner. In mice the dose was given orally for 6 days/week for 4 weeks. A network pharmacology-based approach and molecular docking were also utilized to predict the target genes and the underlying mechanism of ETV in Hb-F induction. ETV in a dose dependent significantly increased the level of total haemoglobin, gamma-globin mRNA expression, and Hb-F production in the K562 cell line. In β-YAC mice, ETV confirmed gamma-globin gene expression at the mRNA and protein levels, as evidenced by increased haemoglobin F production in red blood cells. Furthermore, the docking analysis indicated that NFKB, PTGS2, HDAC2, and HIFIA are potential key targets of ETV, possibly involved in Hb-F induction, and must be validated experimentally. ETV, an approved drug for antiviral activity, increased the level of Hb-F in K562 cell line and in β-YAC transgenic mice, may suggest its use for the management of β-thalassemia. However, further research is required to test the drug's efficacy and the underlying gene targets in CD34+ erythroid progenitors of β-thalassemic patients.
BACKGROUND:Elexacaftor-tezacaftor-ivacaftor (ETI) has transformed cystic fibrosis (CF) care, but individual responses remain variable. Although longitudinal studies indicate that ETI reshapes lung and gut microbiota, the contribution of direct drug-microbe interactions remains unclear. We used an in vitro pharmacomicrobiomics approach to screen bidirectional interactions between individual cystic fibrosis transmembrane conductance regulator (CFTR) modulators and CF-relevant bacteria from pulmonary and intestinal niches. METHODS:Thirty-five Gram-positive and Gram-negative strains (15 reference strains, 20 pulmonary clinical isolates) were selected according to their frequency in CF lung and gut microbiota. Bacteria were grown in brain-heart infusion or M9 medium with or without elexacaftor, tezacaftor or ivacaftor. Growth was assessed by OD600 and CFU counts, and residual parent-drug concentrations were quantified by validated liquid chromatography-tandem mass spectrometry. Logistic growth models were fitted to OD600 data. RESULTS:Elexacaftor modestly increased growth of Pseudomonas aeruginosa and Escherichia coli reference strains, but not clinical P. aeruginosa isolates, suggesting strain-dependent effects. Ivacaftor inhibited all Staphylococcus aureus strains, including methicillin-resistant S. aureus, and several anaerobic commensals, while leaving P. aeruginosa and E. coli largely unaffected. Tezacaftor had no detectable effect on growth. In sterile media, all three modulators remained relatively stable. In bacteria-exposed cultures, parent-drug concentrations decreased by approximately 60-70%, with several pathogens and commensals showing high-depleting phenotypes. CONCLUSIONS:CFTR modulators and CF-associated microbiota interact bidirectionally in vitro. Elexacaftor and ivacaftor exert compound-, species- and strain-dependent effects on bacterial growth independent of their canonical role in restoring CFTR function, while multiple bacterial species decrease detectable parent-drug concentrations in culture supernatants.
Parkinson’s disease (PD) is increasingly linked to gut-brain axis dysfunction. While broad-spectrum antibiotics may contribute to gut dysbiosis, which has been associated with an increased abundance of curli-producing Enterobacteriaceae and processes implicated in α-synuclein aggregation, certain antibiotic classes, particularly tetracyclines and β-lactams, have demonstrated neuroprotective properties in pre-clinical models of PD. This creates a therapeutic paradox: antibiotics may exert either beneficial or detrimental effects depending on many factors including host microbiome composition. Importantly, much of the current evidence remains observational or pre-clinical, and a direct causal relationship between antibiotic exposure and PD has not yet been established. This review synthesizes current epidemiological, mechanistic, and pre-clinical evidence regarding this paradox and explores the emerging role of nanotechnology in resolving it. Preclinical studies suggest that nanoparticle-based drug delivery systems may enhance BBB penetration and enable more targeted drug release. Such approaches have been proposed as a means to reduce systemic exposure and potentially limit microbiota disruption, although these benefits have not yet been demonstrated in clinical PD populations. We further discuss the translational hurdles, including nanotoxicity and regulatory requirements, and propose a roadmap for future research that integrates metagenomics with precision nanomedicine. Collectively, these findings provide a framework for future investigation of targeted therapeutic strategies for PD. However, substantial mechanistic, clinical, and translational validation is required before their therapeutic potential can be established.
Depolymerized Fucoidan derivatives (DFd) are emerging anticancer agents, yet their relationship with heparanase (HPSE) remains unclear. H2O2-assisted depolymerization of native fucoidan (Fucus vesiculosus) generates DFd characterized by reduced number- and weight-average molecular masses (Mn and Mw) and degree of sulfation (DS) with oxidative structural modifications, resulting in heterogeneous yet bioactive compounds with reduced anticoagulant activity compared to the native fucoidan. Their inhibitory effect on HPSE in vitro was associated with both Mn and DS, although no dominant factor was identified. Biologically, DFd decreased mitochondrial metabolic activity and migration in breast cancer models according to Mn and DS reductions, with differential responses between MCF7 and MDA-MB-231 cells. Despite minor cell line-specific changes in HPSE expression, heparan sulfate (HS) levels remained unchanged, suggesting that these effects are not driven by altered HPSE expression or extracellular matrix degradation. This study provides a key original contribution by tracking DFd cellular internalization for the first time in these cell lines, revealing a dependence on Mn/DS and a correlation with the observed migration inhibition for the MCF7 cells treated with the smallest DFd. These findings highlight cellular uptake as a critical determinant of fucoidan bioactivity and open new perspectives for understanding and optimizing their anticancer mechanisms.
Engineered albumin interfaces can modify the biological identity and performance of non-albumin nanoparticles in cancer nanomedicine, but their independent contribution is often obscured by multicomponent designs. This critical review evaluates studies in which albumin or an albumin-derived construct was deliberately positioned at the outer interface of a non-albumin nanoparticle or drug nanocrystal before biological exposure. Architectures included adsorbed layers, covalent or crosslinked coatings, albumin-stabilized nanocrystal interfaces, and hybrid or modified constructs. Across 35 primary studies, albumin served mainly as a stabilizing, cargo-binding, release-modulating, ligand-bearing, or biologically active interfacial component. Matched controls supported selected albumin-associated changes in colloidal stability, cellular interaction, tumor drug delivery, macrophage uptake, and secondary-corona composition. By contrast, imaging, photothermal and radiation responses, pharmacokinetics, biodistribution, and antitumor efficacy often depended on the nanoparticle core, payload, targeting ligand, other surface components, or applied energy and therefore generally represent formulation-level outcomes unless albumin was specifically isolated. Secondary-corona studies suggest that engineered albumin interfaces may reshape corona composition rather than suppress protein adsorption, but the biological consequences remain incompletely resolved. The persistence and conformational accessibility of manufactured albumin interfaces during circulation are also poorly defined, and proposed receptor-mediated mechanisms rarely have direct support. Corona-proteomic findings remain hypothesis-generating and have not established clinically validated cancer detection or diagnostic performance. Future translation requires matched albumin-free and protein-replacement controls, independent in vivo tracking of nanoparticle cores and albumin interfaces, standardized secondary-corona characterization, pharmacokinetic and safety evaluation, and validation of promising bovine serum albumin (BSA)-based systems using clinically relevant human serum albumin (HSA) interfaces.
This study explores the protective effects and mechanistic pathways of imeglimin in models of lipopolysaccharide (LPS)-induced acute lung injury (ALI). Imeglimin administration significantly enhanced survival rates, mitigated pulmonary edema, and attenuated histopathological lung damage in murine subjects. Furthermore, imeglimin decreased inflammatory cell infiltration, reduced protein leakage, lowered oxidative stress markers such as malondialdehyde, suppressed pro-inflammatory cytokines including IL-6 and CXCL2, and elevated the activity of the antioxidant enzyme superoxide dismutase within bronchoalveolar lavage fluid. In vitro studies using human pulmonary microvascular endothelial cells revealed that imeglimin preserved mitochondrial integrity and membrane potential, diminished reactive oxygen species production, and inhibited the opening of the mitochondrial permeability transition pore. Transcriptomic analyses identified TXNIP as a critical gene downregulated by imeglimin via enhanced methylation of its promoter region. This epigenetic modulation correlated with increased expression and activity of DNMT3B, and decreased activities of TET3. Molecular docking studies further substantiated the direct binding of imeglimin to DNMT3B and TET3, suggesting that imeglimin facilitates TXNIP promoter methylation by modulating interactions among DNMT3B, TET3, and the TXNIP promoter. Comparative analyses with seven other hypoglycemic agents demonstrated that imeglimin conferred the lowest mortality rate and hypoglycemia incidence in ALI mice, underscoring its superior therapeutic efficacy and safety. Collectively, these results indicate that imeglimin confers protection against LPS-induced ALI by promoting TXNIP promoter methylation, silencing TXNIP expression, enhancing mitochondrial quality control, and attenuating oxidative stress and inflammation, thereby improving survival outcomes and positioning imeglimin as a promising therapeutic candidate for ALI management.
Acne vulgaris is a chronic inflammatory skin condition that affects everyone at least once in their life. Notably, acne vulgaris predominantly negatively affects the quality of life of teenagers. Treating acne vulgaris remains challenging for some patients. Currently, a variety of topical and oral medications are used to eliminate acne vulgaris; however, risks involving bacterial drug resistance and potential side effects persist. Therefore, demand for novel alternatives to conventional medical treatments is growing. Photothermal therapy (PTT) has the potential to provide quick, safe, effective, and side-effect-free acne vulgaris treatment. PTT using nanoparticles (NPs) converts near-infrared (NIR) light into heat to target and deactivate overactive sebaceous glands, leading to reduced sebum production and inflammation. A handful of human clinical studies on NP-mediated PTT showing improvements in skin clarity and patient satisfaction have been conducted. However, these trials involved small patient populations, short durations, and a lack of sufficient controls. Artificial intelligence (AI) can revolutionize acne treatment through a synergistic approach that incorporates PTT. The combination of AI with NP-mediated PTT enables rapid and large-scale patient treatment, precise identification of acne lesions, and real-time monitoring of treatment progress. This synergy can save time, improve diagnostic accuracy, and optimize treatment options for patients. In this review, we summarize and discuss recent advances in clinical studies of NP-mediated photothermal acne therapy integrated with AI, which holds promise for healing hundreds of millions of patients with acne.
Liver fibrosis links chronic liver injury to cirrhosis, hepatic decompensation, and hepatocellular carcinoma (HCC), while current non-invasive tests mainly estimate fibrotic burden and incompletely capture active stromal biology. Fibroblast activation protein-α (FAPα), a cell-surface serine protease enriched in activated hepatic stellate cells (HSCs), myofibroblasts, and cancer-associated fibroblasts (CAFs), is a biologically informative and pharmacologically tractable stromal target because of its extracellular accessibility and compatibility with ligand-based imaging, protease-activated drug delivery, radionuclide therapy, and cellular therapeutic platforms. This review evaluates FAPα as a candidate druggable stromal hub across the fibrosis-HCC continuum while distinguishing FAPα expression, enzymatic activity, FAPI tracer uptake, and causal biological function. We summarize its membrane topology, dimerization, catalytic pocket, dipeptidyl peptidase and endopeptidase activities, and structural basis for selective targeting within the S9B serine protease subfamily. We discuss chronic liver injury-induced FAPα-positive HSC programs, extracellular matrix (ECM) remodeling, integrin-mediated mechanotransduction, and macrophage-stromal inflammatory amplification. At the fibrosis-cirrhosis-HCC interface, FAPα-positive CAF states are associated with, and in selected experimental models contribute to, spatial tumor architecture, immune exclusion, and therapeutic resistance. We further evaluate tissue profiling, investigational FAPα-responsive magnetic resonance imaging (MRI), early-clinical fibroblast activation protein inhibitor (FAPI)-based positron emission tomography (PET), fluorescence-guided surgery, direct enzymatic inhibition, FAPα/integrin co-targeting, protease-activated prodrugs, targeted nanomedicine, radioligand theranostics, anti-FAP cellular therapy, and combinations with immunotherapy or anti-angiogenic treatment. Collectively, FAPα is an evidence-stage-dependent stromal target for mechanistic stratification, molecular imaging, and therapeutic development; prevention of HCC and reduction of recurrence remain hypotheses requiring prospective validation.
Background Cannabidiol (CBD) shows promising anti-cancer effects, including reducing proliferation and migration and inducing cell death. However, its impact on cancer gene regulation and epigenetic mechanisms remains poorly understood, particularly in prostate cancer, a disease characterised by widespread epigenetic alterations. Purpose This study investigated whether CBD exerts anti-cancer effects in prostate cancer by modulating cell viability, gene expression, and the epigenome and whether it enhances the efficacy of targeted and hormonal agents. Study design/methods Prostate cancer cell lines (DU145, PC3, LNCaP) were treated with varying concentrations of CBD, Talazoparib, GSK126 and enzalutamide. Cell viability was assessed by MTT. Transcriptomic changes were analysed by RNAseq and qRT-PCR, and epigenetic effects were evaluated by the Infinium MethylationEpic V2.0 BeadChip array. Analysis of total 5mC and expression/activity of EZH2 were assessed by ELIZA and Western blot respectively. Results: Epigenetically, CBD altered methylation patterns in LNCaP cells, while modulating DNMT1 and EZH2 expression across models. However, EZH2 catalytic activity was unchanged. CBD induced widespread transcriptional changes, particularly in LNCaP cells, with enrichment of cell cycle pathways and downregulation of key regulators (e.g., CDK1/2). CBD reduced cell viability in a dose-dependent manner, showing additive or synergistic effects with both targeted and hormonal therapies. Conclusion CBD exerts anti-cancer effects in prostate cancer, potentially through disruption of cell cycle regulation and epigenetic modulation. These findings support its potential as a therapeutic agent, particularly in combination with targeted treatments.
HIV-1 persists in cellular reservoirs of latent virus that resist antiretroviral therapy (ART) and immune clearance, necessitating novel therapeutic strategies. While "shock-and-kill" approaches aim to reactivate latent virus under immune pressure, this strategy has demonstrated limited clinical efficacy. In contrast, "block-and-lock" strategies that reinforce and maintain HIV-1 quiescence represent a complementary approach to prevent reservoir expansion and enable long-term ART-free remission. We previously demonstrated that EGCG, the major phenolic compound of green tea, reverses HIV-1 latency by inhibiting UHRF1 expression, a key regulator involved in the epigenetic silencing of HIV-1. However, EGCG exhibits poor bioavailability and dose-dependent cytotoxicity, which severely limit its clinical use. Here, we evaluated (+)-catechin:lysine 1:2, a more stable and bioavailable polyphenol complex with unexplored anti-HIV properties. In J-Lat cell subclones, high concentrations (50-300 μg/mL) of (+)-catechin:lysine 1:2 induced HIV-1 reactivation to levels comparable to those induced by EGCG, but without dose-dependent cellular toxicity. Mechanistically, HIV-1 reactivation operated through UHRF1-independent pathways, distinguishing (+)-catechin:lysine 1:2 from EGCG. Additionally, sequential vitamin C pre-treatment at a 1:2 molar ratio significantly potentiated catechin-mediated reactivation, whereas concurrent co-treatment antagonized the effect, demonstrating that precise dosing time schedules critically determined efficacy. Strikingly, short-term exposure (24 h) of therapeutically achievable low doses of (+)-catechin:lysine 1:2 (10 μg/mL) paradoxically promoted HIV-1 quiescence, suppressing spontaneous viral reactivation. Importantly, this quiescence-promoting effect was confirmed in cells isolated from ART-treated people with HIV (PWH). These findings position of (+)-catechin:lysine 1:2 as a safely administrable and bioavailable molecule with promising potential for "block-and-lock" anti-HIV-1 cure strategies.
Obesity is a major risk factor for colorectal cancer (CRC), promoting tumor initiation through chronic inflammation and metabolic dysregulation. Cancer stem cells (CSCs) drive CRC progression through Notch1, Wnt/β-catenin (Wnt), and PI3K/mTOR (mTOR) signaling. Omega-3 polyunsaturated fatty acids (EPA and DHA) and epigallocatechin-3-gallate (EGCG) may modulate these pathways. This study investigated the effects of obesogenic-like stimuli on CSC-related CRC signaling and the preventive potential of EPA:DHA-EGCG in patient-derived organoids (PDOs) from normal mucosa (NM) of non-obese (non-OB) patients. CD133 and CD44 expression was evaluated in tumor tissues and matched NM from obese (OB) and non-OB CRC patients. PDOs derived from NM of non-OB patients were exposed to an adipocyte-conditioned medium reproducing an obesogenic adipokine profile, with or without EPA:DHA (1:1) plus EGCG (EDE). Viability, morphology, and molecular markers of stemness, differentiation, and CRC-related pathways were assessed. OB CRC tissues exhibited a CSC change from CD133-high/CD44-low to CD44-high/CD133-low compared with non-OB CRC. In PDOs, OB-EDE recapitulated this CD44-high/CD133-low phenotype, increasing organoid viability and size while maintaining KRT20 comparable to control (CTRL). HES1 was significantly upregulated, consistent with modulation of Notch1 signaling. OB-EDE downregulated Wnt target genes and increased the mTOR downstream effector p-S6R. OB + EDE exerted marker- and pathway-specific effects, maintaining PROM1 at CTRL levels, reducing CD44 and HES1, and increasing KRT20, while LGR5 remained suppressed. Compared with OB-EDE, OB + EDE further reduced the expression of Wnt target genes and reduced p-S6R to CTRL, whereas p-mTOR/mTOR was not significantly affected. Overall, obesogenic-like stimuli were associated with a CD44-high stem-like phenotype accompanied by coordinated changes in Notch1, Wnt and mTOR signaling. EDE exerted differential effects on these molecular alterations, supporting its potential as a complementary bioactive strategy in obesity-associated CRC. Pharmacological inhibition using DAPT provided preliminary functional support for a contribution of Notch1 signaling to the maintenance of this obesogenic phenotype.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive dysfunction and impaired synaptic plasticity. This study evaluated whether chronic L-carnitine (LC) attenuates amyloid-beta 1-42 (Aβ₁-₄₂)-induced spatial learning and memory deficits following intracerebroventricular (ICV) Aβ₁-₄₂ infusion. Forty adult male Wistar rats were randomly assigned to four groups (n = 10/group): Sham + Vehicle, Sham + LC, AD + Vehicle, and AD + LC. Following ICV Aβ₁-₄₂ infusion, LC (100 mg/kg/day, i.p.) or vehicle was administered for 28 days. Spatial learning and memory, hippocampal long-term potentiation (LTP), oxidative stress markers, Aβ plaque density, and neuronal integrity were evaluated. Compared with the AD + Vehicle-treated rats, LC attenuated Aβ₁-₄₂-induced impairments in spatial learning and memory, as indicated by reduced escape latency on training days 3-4 (both p < 0.01) and increased time spent in the target quadrant (p < 0.01). LC also improved hippocampal LTP, as reflected by a 58.4% increase in population spike (PS) amplitude potentiation (p < 0.01). Additionally, LC increased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities by 40.3%, 34.4%, and 37.4%, respectively, while reducing malondialdehyde (MDA) levels by 22.0% (all p < 0.01). LC also decreased Aβ plaque density by 36.6% and increased intact pyramidal neurons by 58.3% (both p < 0.01). These findings indicate that LC mitigates Aβ₁-₄₂-induced spatial learning and memory deficits and is associated with improved hippocampal LTP, enhanced antioxidant enzyme activities, reduced lipid peroxidation and Aβ plaque density, and preserved neuronal integrity.
BACKGROUNDS:Chronic hypoxia-associated pulmonary hypertension (PH), especially COPD-related PH with right heart failure, is a recalcitrant clinical challenge as current therapies fail to effectively halt its progression. Olprinone (Olp), a phosphodiesterase III inhibitor with well-defined cardiovascular regulatory effects, has unclear roles and molecular mechanisms in hypoxia-induced PH. METHODS:This study combined clinical, preclinical and cellular approaches: a retrospective analysis of 24 COPD-related PH patients (13 with Olp plus standard therapy, 11 with standard therapy plus other inotropes); a chronic hypoxia-induced PH (CHPH) rat model (10% O₂ for 4 weeks) treated with intraperitoneal Olp (0.2/0.4 mg/kg/d) for 2 weeks; hypoxic human pulmonary artery smooth muscle cells (HPASMCs, 3% O₂) exposed to Olp (30-300 nM), with IDH1 function validated by overexpression. RESULTS:Clinically, Olp significantly reduced pulmonary artery systolic pressure (PASP) and serum NT-proBNP, and elevated the TAPSE/PASP ratio (a core right ventricular-pulmonary arterial coupling marker, all P < 0.05), with no significant changes in 6-minute walk distance or pulmonary function. In rats, Olp dose-dependently decreased mean PAP, RVSP and pulmonary artery medial thickening, and downregulated collagen I, α-SMA and PCNA (all P < 0.05 vs. hypoxia group). Olp suppressed hypoxic HPASMC proliferation, migration and invasion (P < 0.05), reduced IDH1 expression, and IDH1 overexpression completely reversed Olp's protective effects on HPASMCs. CONCLUSION:Our findings suggest olprinone improves key right ventricular-pulmonary arterial coupling and pulmonary vascular remodeling in hypoxia-associated PH models, likely via regulating IDH1-related pathways. Clinical observations remain preliminary; further studies with COPD-PH models and prospective clinical trials are needed to confirm its therapeutic value.
