Glioblastoma is a highly malignant and refractory brain tumor with limited treatment options and poor prognosis. Although antibody-drug conjugate (ADC) have shown potential in glioblastoma therapy, their clinical utility is hampered by off-target toxicity. This study aimed to develop a novel NAD(P)H:quinone oxidoreductase 1 (NQO1)-responsive ADC, NRBM-25, to enhance targeting specificity and safety, and to investigate its antitumor mechanisms. NRBM-25 was constructed using an NQO1-enzyme-responsive linker to conjugate a vascular endothelial growth factor-targeting antibody with the cytotoxic payload monomethyl auristatin E, enabling tumor-specific activation. The NQO1-dependent activation mechanism was validated in isogenic and native cell models. In vitro and in vivo, NRBM-25 exhibited potent and selective cytotoxicity against NQO1-high glioblastoma cells while sparing NQO1-low normal cells. The conjugate induced dual modes of cell death-apoptosis and ferroptosis-accompanied by mitochondrial dysfunction, oxidative stress, and cell cycle arrest. In animal models, NRBM-25 significantly inhibited tumor growth, prolonged survival with minimal systemic toxicity, and outperformed temozolomide at medium-to-high doses. This approach of using an enzyme-dependent linker to dually induce apoptosis and ferroptosis offers a potential strategy to overcome treatment resistance, representing a new exploration in ADC-based therapy for glioblastoma. SIGNIFICANCE STATEMENT: A novel antibody-drug conjugate targeting glioblastoma was developed, which uses a tumor-specific enzyme activation mechanism and demonstrates favorable safety and antitumor efficacy in preclinical models. This candidate represents a new exploration in antibody-drug conjugate-based therapy for glioblastoma.

Although the clinical efficacy of apatinib in advanced gastric cancer has been widely confirmed, its anticancer mechanism still needs to be further studied. This study found that apatinib significantly inhibited gastric cancer cell viability by inducing the expression of cysteine dioxygenase 1 (CDO1; a key ferroptosis regulatory gene) and then enhancing ferroptosis, and CDO1 knockdown reduced ferroptosis. Apatinib and CDO1 significantly downregulated the expression of glutathione peroxidase 4 and solute carrier family 7 member 11, reduced cell viability and glutathione production, promoted acyl-CoA synthetase long chain family member 4 expression, and increased the accumulation of reactive oxygen species, Fe[2+], and malondialdehyde; when knocking down CDO1 expression or activating phosphoinositide 3-kinase (PI3K)/Akt pathway, the expression of key ferroptosis regulatory genes and the contents of ferroptosis markers were significantly restored, indicating that apatinib-induced ferroptosis is associated with CDO1/PI3K/Akt axis. Meanwhile, both apatinib and CDO1 significantly inhibited the expression and secretion of metalloproteinase (MMP)-2 and MMP-9, and reduced cell invasion and migration; the ferroptosis inhibitor Fer-1, CDO1 small interfering RNA, and PI3K/Akt pathway activator 740 Y-P all weakened the regulation of apatinib and CDO1 on MMP expression, cell invasion and migration. Further in vivo nude mouse tumorigenesis experiments showed that apatinib significantly inhibited tumor growth and the expression of glutathione peroxidase 4 and solute carrier family 7 member 11, promoted CDO1 and acyl-CoA synthetase long chain family member 4 expression, increased reactive oxygen species, Fe[2+], and malondialdehyde contents, and reduced glutathione production; after knocking down CDO1 expression, all these ferroptosis indicators were significantly restored. Therefore, apatinib inactivates PI3K/Akt pathway by promoting CDO1 expression, thereby enhancing ferroptosis and then inhibiting tumor growth. SIGNIFICANCE STATEMENT: Apatinib is a small-molecule tyrosine kinase inhibitor and can inhibit tumor cell growth through various mechanisms other than antiangiogenesis. Therefore, understanding the anticancer mechanism of apatinib is of great significance for its clinical application.

Autoimmunity plays a central role in hepatobiliary and pancreatic diseases; however, the mechanisms underlying the initiation and progression of autoimmune cholangitis and pancreatitis remain incompletely understood. Glycine, a nutritionally nonessential amino acid, has immunomodulatory and hepatoprotective properties. This study investigated the effects of dietary glycine on innate and adaptive immune responses induced by double-stranded RNA. Chronic administration of polyinosinic-polycytidylic acid for 24 weeks induced cholangitis- and pancreatitis-like phenotypes, characterized by elevated serum alkaline phosphatase and amylase levels, increased levels of autoantibodies, including antimitochondrial M2 and anti-lactoferrin antibodies, and periductal infiltration of macrophages, T cells, and B cells. These changes were accompanied by the induction of proinflammatory cytokines and type I interferons. Dietary glycine supplementation significantly attenuated these pathological features in both preventive and therapeutic settings. Glycine suppressed early innate immune activation, as evidenced by reduced hepatic expression of tumor necrosis factor α, interleukin (IL)-1β, IL-6, IL-12b, and type I interferons. Furthermore, glycine modulated downstream adaptive immune responses, including suppression of T helper (Th)2-related cytokines (IL-4 and IL-13), the Th17-related cytokine IL-23a, and the regulatory cytokine IL-10, along with reduced autoantibody production. These findings suggest that glycine acts upstream in the inflammatory cascade by attenuating innate immune signaling that drives downstream adaptive immune responses. In conclusion, dietary glycine suppresses double-stranded RNA-induced innate and autoimmune responses in the liver and pancreas, supporting its potential as a safe, mechanism-based therapeutic strategy for autoimmune hepatobiliary and pancreatic diseases. SIGNIFICANCE STATEMENT: Dietary glycine suppresses double-stranded RNA-triggered innate immune activation and autoantibody production, preventing autoimmune-like cholangitis and pancreatitis. By modulating upstream toll-like receptor 3/pattern-recognition receptor and type I interferon pathways, glycine provides broad yet pharmacologically safe immunoregulation with potential relevance to autoimmune hepatobiliary and pancreatic diseases such as primary biliary cholangitis and autoimmune pancreatitis.

β-Adrenergic receptors (βxAR) are G protein-coupled receptors with major physiological and therapeutic relevance in cardiovascular and respiratory conditions. Although numerous ligands have been investigated, binding affinities and functional annotations remain incomplete and fragmented across databases. This review systematically integrates ligand binding affinities and functional profiles at recombinant human βxAR from the Psychoactive Drug Screening Program Ki database and the International Union of Basic and Clinical Pharmacology/British Pharmacological Society (IUPHAR/BPS), with a focus on clinically important ligands, receptor selectivity, and characteristic patterns. Database agreement, affinity patterns, stereoisomer-specific information and concordance with primary literature are evaluated. In total, 156 ligands with pKi ≥ 5 were collected. Median pKi values were 6.6 for hβ1AR, 6.15 for hβ2AR, and 5.8 for hβ3AR. A pronounced overlap between hβ1AR and hβ2AR was observed, reflected in similar median affinities and shared ligand profiles, whereas hβ3AR showed narrower data coverage and a more distinct affinity distribution. Most ligands were listed for more than 1 hβxAR, and 37 ligands had binding data for all 3 hβxAR. Functional characterization was available for 90 ligands, whereas 61 lacked annotations. Database comparison revealed low overlap in ligand listings (28.8%). Stereoisomer data were highly incomplete, and in several cases, affinity-based stereoisomer comparisons differed from literature-reported stereoisomeric superiority. Summarized, substantial similarity between hβ1AR and hβ2AR ligand binding patterns is observed, whereas hβ3AR appears less extensively characterized. Considerable fragmentation, incomplete receptor profiling and methodological heterogeneity limit the evaluation of receptor and ligand characteristics. Binding affinity and functional behavior must be interpreted as context-dependent parameters influenced by experimental systems, G protein coupling conditions and stereochemical specification. SIGNIFICANCE STATEMENT: This analysis systematically integrates ligand binding affinities and functional labels at recombinant human βxAR from the Psychoactive Drug Screening Program (PDSP) Ki database and the International Union of Basic and Clinical Pharmacology/British Pharmacological Society (IUPHAR/BPS) Guide to Pharmacology, providing a structured and comprehensive overview of ligand profiles. It identifies substantial data fragmentation, incomplete receptor and stereoisomer characterization, and limited concordance between databases. For clinically relevant ligands, it was demonstrated that therapeutic subtype classification does not always correspond to strict affinity-based receptor selectivity.

Twinfilin-1 (TWF1) is a cellular protein that binds actin, phosphoinositides, and capping protein. Although multiple studies have reported the tumor-regulatory role of TWF1, a comprehensive and critical review of these findings remains unavailable. This review addresses this gap by consolidating current evidence on the tumor-modulatory functions of TWF1. The literature indicates that dysregulation of TWF1 or its actin-binding activity can promote carcinogenesis by influencing interleukin-11/signal transducer and activator of transcription 3, protein kinase B, epithelial-to-mesenchymal transition, or autophagy. The mechanisms by which TWF1 regulates tumorigenesis are tissue-dependent and vary across different tissues. At least 10 microRNAs, 2 long noncoding RNAs, and 1 circular RNA have been identified as interacting with TWF1, either directly or indirectly, to regulate its expression and tumor-controlling function. High TWF1 expression has been observed in at least 8 types of solid tumors, but not in liquid cancers, highlighting its potential as a diagnostic biomarker for solid tumors. Furthermore, its increased expression in solid tumors can help predict cancer patient prognosis. However, despite these findings, the translational potential of TWF1 requires further validation through in-depth mechanistic studies and large-scale clinical trials, as current evidence is primarily derived from bioinformatics analyses, preclinical studies, or small prospective cohorts. Once the tumor-promoting role of TWF1 is confirmed, future research should then evaluate its sensitivity and suitability as a biomarker or therapeutic target. SIGNIFICANCE STATEMENTS: Twinfilin-1 (TWF1) regulates various cellular processes that contribute to solid cancer cell proliferation, metastasis, and resistance to treatment. These functions suggest that TWF1 has translational potential as both a biomarker and a therapeutic target in cancer management.

A growing body of literature suggests that integrase strand transfer inhibitors (INSTIs), a cornerstone of antiretroviral therapy for the treatment of HIV infection, seem to possess antiproliferative properties against some types of cancer and/or anti-invasion effects. Preclinical studies describe as INSTIs, including dolutegravir, raltegravir, cabotegravir, and elvitegravir and some 1,2,3-triazole derivatives, can inhibit the growth of various cancer cell lines and some of them seem to also have direct inhibitory effects on human endogenous retrovirus reactivation. This review comprehensively synthesizes and analyzes this evidence, detailing the proposed multimechanistic actions behind these effects, such as induction of DNA damage, inhibition of key oncogenic pathways, triggering of oxidative stress and diverse cell death modalities. All described compounds seem to have potential effects on cancerous cells, and these effects may be also independent from their antiretroviral activity, that is, may be separated from the mere effect on suppression of human endogenous retrovirus reactivation in cancerous cells. Although the type of target cancer, the mechanisms of action (ie, enzyme or protein inhibition, oxidative stress, DNA methylation, and DNA damage) or the ultimate effects (ie, cell proliferation, migration, or invasion) were different in most of the studies and sometimes inconsistent, all these studies suggest several potential targets of INSTIs that deserve further investigations. Although the incidence of many cancers is higher in people with HIV infection compared with the general population, the relative risk has been decreasing over time for many cancers. Further studies are then needed to assess the effects of INSTI therapies on malignancies in people living with HIV. SIGNIFICANCE STATEMENT: Currently, the effects of using integrase strand transfer inhibitors (INSTIs) on cancer development in people living with HIV are difficult to determine because of the multifactorial cause of cancer in these patients and the long period in which cancer manifests. This review examines the antiproliferative properties of INSTIs, describing the proposed mechanisms through which they may exert this effect in preclinical studies.

Morphine remains a cornerstone for managing nociceptive pain, yet its clinical utility is severely limited by the development of tolerance and physical dependence. This study investigated the potential of pyrroloquinoline quinone (PQQ), a natural redox-active cofactor with antioxidant and anti-inflammatory properties, to mitigate these adverse effects in mice. Animals received morphine (5 mg/kg, i.p.) twice daily for 9 days. PQQ (5 mg/kg, i.p.) was administered either concurrently with morphine for 9 days or as a single dose on the final day. Results demonstrated that PQQ treatment significantly reduced the development of morphine tolerance, as measured by the hot plate test, and dependence through reduction of naloxone-precipitated withdrawal signs. Biochemically, PQQ decreased levels of key mediators involved in neuroplasticity and oxidative stress, including calcium/calmodulin-dependent protein kinase II, glutamate, tumor necrosis factor-α, and malondialdehyde, while increasing the antioxidant glutathione. Immunohistochemical analysis showed that PQQ reduced c-Fos protein expression in the brain. Furthermore, the docking analysis supported these findings and uncovered novel underexplored molecular targets, with alternative distinct binding modes and modulatory effects. PQQ displayed strong binding affinities for N-methyl-D-aspartate receptors at a previously unrecognized site, as well as for calcium/calmodulin-dependent protein kinase II, IκB kinase, and Kelch-like ECH-associated protein 1 proteins. These interactions suggest distinct modulatory effects on critical pathways involved in tolerance and dependence. In conclusion, PQQ effectively reduces morphine-induced tolerance and dependence through multifaceted mechanisms. Its efficacy, combined with its high safety profile as a natural compound, positions it as a promising therapeutic candidate for overcoming the significant challenges associated with long-term morphine use. SIGNIFICANCE STATEMENT: This study identifies the neuroprotective effect of PQQ against morphine tolerance and dependence, where PQQ affected many targets, with unique interaction sites and binding modes to N-methyl-D-aspartate receptors and calcium/calmodulin-dependent protein kinase II. By targeting relevant pathways, PQQ can be considered as a novel effective suppressor of morphine tolerance and dependence, and a safe natural compound that offers a multifaceted promising approach for this problem.

The aryl hydrocarbon receptor (AHR) is a ligand activated transcription factor that has emerged as a key modulator of several physiological and pathological processes. Historically, AHR has been studied for its role as a mediator of the toxic responses of environmental pollutants, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin. Because of this, its potential as a therapeutic target was overlooked. AHR is now regarded as a multifunctional regulator of inflammation, immunity, barrier tissue integrity, metabolism, and cancer biology. AHR signaling is modulated by an array of structurally diverse endogenous, microbial, dietary, pharmaceutical, and xenobiotic ligands. AHR exhibits extensive crosstalk with many signaling pathways, which contributes to highly context-specific biological outcomes. Recent advances and enhanced interest in AHR pharmacology have accelerated the development of therapeutically relevant AHR ligands, including the clinically approved AHR agonist, tapinarof, for the treatment of psoriasis and atopic dermatitis, as well as emerging AHR antagonist strategies in cancer immunotherapy. Structural efforts have succeeded in providing valuable insight into ligand-AHR interactions that will further contribute to improved and rational design of AHR ligands. However, significant challenges remain, including AHR ligand promiscuity, complex negative feedback regulation, extensive signaling crosstalk, and long-term safety concerns. Here, we review the molecular mechanisms of AHR activation, its physiological and pathological functions, and the current advances of its therapeutic targeting. SIGNIFICANCE STATEMENT: The aryl hydrocarbon receptor (AHR) has emerged from its origins in toxicology to become a promising clinically relevant target for the treatment of dermatological, gastroenterological, and autoimmune diseases as well as cancer. This review summarizes the current knowledge of AHR pharmacology, including molecular signaling, endogenous and exogenous ligands, physiological and pathological functions, present and emerging therapeutic targeting strategies, as well as challenges that remain for translation into safe clinical therapies.

Polypharmacy, the concurrent use of multiple medications, is increasingly prevalent in older people and is associated with adverse outcomes such as falls, frailty, functional, and cognitive decline, and increased hospitalization and mortality. The liver, as the primary site of metabolism, is exposed to varying drug concentrations during first pass metabolism, hepatic clearance and perfusion, potentially causing alterations in liver sinusoidal endothelial cells (LSEC). LSEC are specialized endothelial cells responsible for maintaining fenestrations-dynamic, transcellular pores that facilitate the exchange of substances between the blood and liver parenchyma. Disruption of fenestrations can compromise liver function, contributing to a variety of hepatic disorders. This study investigated the effects of 4 commonly prescribed drugs-metoprolol, citalopram, oxybutynin, and oxycodone-on LSEC function. We examined their impact on LSEC viability, endocytosis, and fenestration morphology at both systemic steady-state and first-pass concentrations, separately and in a polypharmacy cocktail to model clinical exposure. All treatments induced sublethal metabolic changes, but effects on LSEC functions were drug- and concentration-dependent. Citalopram and oxybutynin caused dose-dependent defenestration, whereas metoprolol and oxycodone produced mild, nondose-dependent effects. Endocytic activity was increased with oxybutynin, metoprolol, oxycodone, and the polypharmacy cocktail, whereas citalopram had no effect. The polypharmacy cocktail triggered synergistic defenestration at first-pass concentrations, but not at steady-state levels. These results highlight the concentration-dependent and combinatorial effects of polypharmacy on LSECs, emphasizing the need to consider endothelial responses in drug safety and pharmacokinetic assessments, particularly in patients exposed to multiple medications. SIGNIFICANCE STATEMENT: Polypharmacy is increasingly common, however, its effects on liver sinusoidal endothelial cells remain poorly understood. Using in vitro models, this study shows that drug combinations can induce sublethal functional and structural alterations in these cells, highlighting endothelial dysfunction as a potential early mechanism contributing to drug-induced liver impairment.

Huntington disease (HD) is a progressive neurodegenerative condition marked by characteristic motor deficits, cognitive decline, and psychiatric disturbances. The mitochondrial toxin 3-nitropropionic acid (3-NP) reliably replicates the HD pathological features by inducing selective striatal and cortical neurodegeneration. Increasing evidence indicates that activation of the Sonic Hedgehog (Shh) signaling could enhance neurotrophic and prosurvival signaling, particularly through modulation of the brain-derived neurotrophic factor (BDNF)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) cascade. Via activation of the Shh pathway, purmorphamine (PUR)-the smoothened agonist-may offer a therapeutic benefit against HD-like pathology. However, its in vivo efficacy in 3-NP neurotoxicity remains unexplored. To examine this possibility, 48 male Wistar rats were assigned to control, PUR, 3-NP, and 3-NP/PUR groups and treated daily for 21 days. Behavioral performance was assessed using the open field, rotarod, novel object recognition, and Morris water maze (MWM) tests, whereas hippocampal, prefrontal, and striatal tissues were evaluated for Shh pathway gene expression, BDNF/Akt/GSK-3β signaling, oxidative/nitrosative stress markers, and histopathological alterations. PUR markedly improved 3-NP induced motor and cognitive deficits, alleviated the BDNF/Akt/GSK-3β signaling, and attenuated oxidative/nitrosative stress, neuroinflammation, and apoptosis. In conclusion, to the best of our knowledge, these findings provide the first evidence that purmorphamine affords significant neuroprotection against 3-NP induced neurodegeneration by activating Shh-dependent BDNF/Akt/GSK-3β signaling and mitigating the key pathological processes, highlighting its promise as a potential therapeutic agent for HD-related neuropathology. SIGNIFICANCE STATEMENT: This study provides the first in vivo evidence that purmorphamine, a Sonic Hedgehog pathway agonist, effectively mitigates 3-nitropropionic acid-induced neurodegeneration by restoring the brain-derived neurotrophic factor/protein kinase B/glycogen synthase kinase-3β signaling cascade. By addressing core pathological features such as motor deficits, cognitive decline, and neuroinflammation, these findings highlight the therapeutic potential of targeting the Sonic Hedgehog pathway as a multitarget strategy for managing Huntington disease-like neurodegenerative conditions.

Signal transducer and activator of transcription 3 (STAT3) is a central oncogenic signaling hub that regulates tumor proliferation, survival, metastasis, angiogenesis, immune evasion, and therapeutic resistance across multiple cancer types. Persistent STAT3 activation, driven by aberrant cytokine signaling, growth factor receptors, and oncogenic kinases, promotes transcriptional programs that support malignant progression and suppress antitumor immunity. Despite its recognized role as an attractive therapeutic target, direct pharmacological inhibition of STAT3 has been challenging because of poor bioavailability, off-target toxicity, and limited tumor specificity of conventional inhibitors. In recent years, nanocarrier-based drug delivery systems have emerged as promising platforms to overcome these limitations by enabling targeted, protected, and sustained STAT3 inhibition. Diverse nanocarrier modalities, including polymeric nanoparticles, lipid-based systems, inorganic nanomaterials, and hybrid or covalent-organic frameworks, have been developed to deliver STAT3 inhibitors, small interfering RNA, short hairpin RNA, and plasmid DNA with improved therapeutic indices. These platforms not only enhance intracellular delivery and tumor accumulation but also enable combination strategies that simultaneously modulate STAT3 and complementary oncogenic pathways. This review provides a comprehensive mechanistic overview of STAT3 signaling in cancer and critically evaluates recent advances in nanocarrier-mediated STAT3 inhibition. Furthermore, it discusses key challenges, including immune context dependency, adaptive resistance, safety concerns, and translational barriers, while highlighting future directions for precision nanomedicine. Collectively, this work underscores the potential of nanotechnology-enabled STAT3 targeting as a next-generation anticancer strategy. SIGNIFICANCE STATEMENT: Persistent activation of signal transducer and activator of transcription 3 represents a critical driver of tumor progression and therapeutic resistance across diverse malignancies, yet its direct pharmacological targeting has remained limited by suboptimal drug properties and systemic toxicity. This review highlights how nanocarrier-based delivery platforms redefine signal transducer and activator of transcription 3 inhibition by improving tumor specificity, intracellular bioavailability, and combinatorial therapeutic potential.

Mast cells (MCs) are versatile, multifunctional immune cells with broad roles in physiological homeostasis and pathogenic processes. MCs are found in most tissues, including skin, lungs, intestines, and peritoneum, and they vary in numbers, types, and biological functions. MCs are implicated in host defense against various pathogens, including bacteria, viruses, and fungi. Additionally, MCs are crucial in protecting against toxins, including those present in venoms from multiple species, such as honeybees, snakes, scorpions, and lizards. Although MCs play an essential role in host defense, they are mostly known for their detrimental actions in allergic reactions, such as asthma, food allergy, anaphylaxis, mastocytosis, and various inflammatory skin conditions. Under such conditions, MCs are activated (via IgE-mediated or other mechanisms) and release a range of potent proinflammatory mediators, including tumor necrosis factor α. In addition to cytokines, they are major producers of histamine and various proteases, including chymase, tryptase, and carboxypeptidase A3. As a result, these mediators contribute to the pathological manifestations associated with inflammatory conditions and other disorders. This review focuses mainly on the biological role of MCs and their proteases, with a focus on chymase and tryptase, as well as their inhibitors as candidate therapies for MC-driven diseases. SIGNIFICANCE STATEMENT: Mast cells (MCs) and their proteases are central regulators of tissue homeostasis, barrier defense, and inflammation across multiple organs, but are also associated with numerous diseases. Ongoing research has shown that the function of MCs is highly dependent on their tissue location, where the local tissue environment shapes their phenotype, protease expression, and, consequently, their biological functions. However, further investigation is required to more precisely understand the physiological conditions governing the transition of MCs from maintaining tissue homeostasis to acquiring pathogenic functions, particularly with respect to their protease-dependent activities. By profiling MC heterogeneity using multiomics approaches to map protease-driven signaling networks, it will be possible to gain deeper insight into their functional roles and establish a conceptual framework to guide the development of next-generation, mechanism-based therapeutics that selectively modulate MC activity in human diseases.