Monoclonal antibodies (mAbs) targeting immune checkpoint pathways such as programmed cell death protein 1 (PD-1)/PD-L1 are central to modern immunotherapy, yet scalable methods to assess their functional blockade remain limited. We present a bead-based flow cytometry assay for quantifying the inhibition of PD-1/PD-L1 interaction by antibodies. Recombinant human PD-1 protein was conjugated to polystyrene beads, and its interaction with recombinant human PD-L1 protein labeled with a fluorochrome was measured. The inhibitory activity of an anti-PD-L1 mAb was quantified based on their ability to disrupt this interaction. The assay was validated for intra- and inter-assay precision, in addition, functionality was confirmed using a T cell coculture assay. The assay demonstrated dose-dependent inhibition by the αPD-L1 mAb, with a calculated mean IC50 of 3.122 µg/mL. The method proved to be reproducible for the determination of antibody blocking activity, with relative standard deviation (RSD) < 20% between three independent runs. At the concentration approximating the IC50 detected on the bead assay, the antibody significantly restored CD69 expression on the T cell surface (p = 0.0001) in a coculture in vitro system. In addition, the methodology could successfully distinguish the blocking capacity of two anti-PD-L1 antibodies with different affinities. This high-throughput compatible platform offers a reliable tool for screening PD-1/PD-L1 blocking antibodies, supporting immunotherapy discovery and development.

Single-stranded DNA gaps (ssDNA gaps) have emerged as a potential indicator of therapeutic response in cancer. Accumulation of ssDNA gaps is associated with increased sensitivity of cancer cells to genotoxic therapies like PARP inhibitors (PARPi) and cisplatin chemotherapy. However, efficient repair or suppression of ssDNA gap formation is associated with therapy resistance and treatment failure. Therefore, understanding how ssDNA gaps form and are repaired can help identify biomarkers that can guide new treatment strategies to overcome resistance. In this review, we discuss different sources of ssDNA gap formation and the repair mechanisms that have been characterized to date. We bring together current knowledge on how these gaps are processed and what their ultimate fate may be. Finally, we discuss how established drugs like PARPi, hydroxyurea, and platinum compounds, induce and/or exploit ssDNA gaps. Throughout this review, we highlight ssDNA gaps as a potential therapeutic vulnerability that can be used to advance personalized cancer therapy.

This study aimed to evaluate the predictive value of the hemoglobin, albumin, lymphocyte, and platelet (HALP) score for pulmonary infections in patients with non-small cell lung cancer (NSCLC) undergoing chemotherapy.

To overcome limitations in melanoma therapy by developing a targeted nanoplatform based on reduced graphene oxide quantum dot (rGOQD) that integrates photothermal therapy (PTT), chemodynamic therapy (CDT), and immune modulation.

In the search for new CDDOs (2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid derivatives) with higher antitumor activity in vivo, thirty CDDO-imidazole derivatives (1-30) were designed and synthesized. Then, 8 was selected due to its superior anti-proliferative activity against three cancer cell lines (B16F10, A549, and HCT116) and its lower toxicity in zebrafish embryos compared to the other evaluated compounds. Further study found that 8 induced apoptosis in HCT116 cells by downregulating Bcl-2, upregulating Bax, and activating caspase-3 to kill cancer cells. Notably, 8 exhibited significant antitumor efficacy comparable to CDDO-Me (bardoxolone methyl), which had entered clinical trials. Taken together, 8 represents a promising candidate for the treatment of cancer and merits further study.

Myeloproliferative neoplasms (MPNs) are clonal hematopoietic disorders driven in large part by aberrant activation of the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway via the JAK2 V617F and related mutations. The success of first-generation ATP competitive JAK2 inhibitors has validated JAK2 as a therapeutic target, yet clinical benefits remain constrained by issues of off target toxicity, limited mutation allele burden reduction, and the emergence of persistence or resistance. In this review, we focus on a rapidly emerging design paradigm: targeting novel binding sites on JAK2 beyond the canonical ATP pocket-including allosteric sites, covalent anchor residues, and pseudokinase regulatory domains. We summarize structural and computational insights into these new sites, compare mechanistic and therapeutic advantages (such as enhanced selectivity, reduced cross JAK inhibition and potential to overcome resistance) and evaluate preclinical and early clinical evidence. We further identify remaining challenges in the development of next generation JAK2 inhibitors-such as site validation, ligand residence time, covalent binding safety, and rational combination therapies-and propose future directions for translation into the MPN clinic. By refocusing the JAK2 inhibitor field around novel binding site exploitation, we suggest a path toward more potent, selective and enduring therapies for MPN patients.

The emergence of immune checkpoint inhibitors (ICIs) has transformed the treatment landscape of metastatic melanoma. However, despite its success, reliable biomarkers for predicting primary resistance are not available in clinical practice. This study seeks to identify predictors of primary resistance based on novel gene expression signatures. The transcriptomic profile of the tumor microenvironment was analyzed using tissue samples from 46 metastatic cutaneous melanoma patients collected prior to the initiation of ICIs therapy. A primary resistance predictive model was trained with the Discovery FFPE RNA-seq subcohort and validated using an independent external cohort of 54 samples. Additionally, liquid biopsy samples from peripheral blood mononuclear cells were analyzed in 8 patients using single-cell RNA sequencing (scRNA-seq) and in 46 patients using flow cytometry. We identified an 82-gene transcriptomic signature composed of tumor- and immune-related genes that stratifies metastatic cutaneous melanoma patients based on primary resistance to ICIs, with key markers including CXCL13, WDR63, MZB1, FDCSP, IGKC and GRIK3. This signature achieved an AUC of 0.814. Immune deconvolution guided by scRNA-seq revealed four immune cell subsets (Plasma cells, Pre-B cells, memory CD4⁺ T cells, and naive CD4⁺ T cells) as prognostic indicators of resistance. We propose a transcriptomic biomarker signature that accurately predicts primary resistance to ICIs in metastatic cutaneous melanoma. Through the integration of immune deconvolution with circulating immune cell profiles, we derived an ImmuneSignature linked to patient survival. By combining these approaches, we provide a framework for enhancing the prediction of immunotherapy outcomes and offer a novel strategy for identifying therapeutic targets to overcome resistance.

Metastatic tumors-secondary malignancies arising from the hematogenous or lymphatic dissemination of cancer cells from primary lesions to distant sites-account for nearly 90% of cancer-associated mortality worldwide. Consequently, studying the effect of epithelial-mesenchymal transition (EMT) on metastatic tumors using experimental data and partial differential equation (PDE) modeling is essential. This study innovatively established a phenotype- and density-regulated chemotaxis coefficient to develop a PDE model characterizing cancer cell migration, proliferation, and EMT, enabling analysis of EMT behavior within the tumor microenvironment and its impact on spreading patterns. Subsequently, incorporating the mechanisms of anti-TGF β RII and cyclophosphamide (CTX), three therapeutic models for tumor metastasis were constructed, with parameter estimation based on experimental data. To predict primary tumor distant metastasis risk, we originally established a tumor metastasis incidence index, thereby evaluating the critical roles of EMT-targeting and cytotoxic chemotherapeutic drugs in tumor progression. Our findings demonstrate that appropriate pharmacological intervention effectively suppresses tumor dissemination, with therapeutic efficacy significantly enhanced upon EMT inhibition. This study establishes a theoretical framework for designing cancer treatment strategies and provides foundational insights for developing personalized therapeutic regimens.

Immune checkpoint inhibitors (ICIs) combined with platinum-based compounds are commonly used in the treatment of certain malignant tumors. This study aims to analyze adverse events (AEs) associated with the combination therapy of ICIs and platinum-based compounds by using the FAERS database.

Myocarditis attributable to immune checkpoint inhibitors is regarded as among the most serious complications of immunotherapeutic treatment. We report a case of gastric cancer who developed myocarditis, and subsequent late complications following treatment with nivolumab and ipilimumab. The patient was managed with corticosteroids, mycophenolate mofetil (MMF), and intravenous immunoglobulin G (IVIG), but later developed supraventricular tachycardia and deep vein thrombosis. This report emphasizes the need to enhance recognition of immune-related toxicities within non-oncology specialties to support prompt interdisciplinary collaboration and appropriate patient management. Additionally, patients can develop myocarditis-related complications even when clinically stable or near the end of treatment, underscoring the need for close and ongoing monitoring.

The purpose of this review is to highlight recently published research that can provide insight into how either sex or chemotherapeutics can impact cancer regulation of muscle anabolic resistance. Critical knowledge gaps are emphasized that are linked to cancer and treatment disruptions to muscle anabolic signaling. We speculate and propose a rationale for estrogen's protective effect against cancer-induced muscle anabolic resistance in females. Furthermore, there is growing evidence that many cancer treatments have the potential to exacerbate muscle anabolic resistance in both males and females. We present current evidence and speculate on how nutritional interventions could serve as key modulators of cancer-induced anabolic resistance in these conditions.

The addition of immune checkpoint inhibitor (ICI) to platinum-based chemotherapy has improved outcomes in patients with advanced non-small cell lung cancer (NSCLC). However, evidence on the efficacy of adding ICI to platinum retreatment in patients who relapse after perioperative platinum-based chemotherapy remains limited.

Phenotypic two-dimensional (2D) high-throughput screening (HTS) is a well-established approach extensively employed in oncological drug discovery by both Academia and the pharmaceutical industry. This methodology has played a pivotal role in the development of a wide range of systemic and targeted therapeutic anticancer agents for clinical use. Recent advances in automation, imaging technologies, and labware design have paved the way for image-based HTS in three-dimensional (3D) cell culture systems. These 3D systems enable the analysis of more physiologically relevant models that closely replicate the characteristics of tumors and their microenvironment. In this study, we present an image-based phenotypic 3D HTS assay utilizing imaging-compatible labware specifically designed to support spheroid formation.

Here, we present a protocol to address the limited intratumoral penetration of small-molecule drugs caused by the dense extracellular matrix (ECM) of pancreatic ductal adenocarcinoma (PDAC). Prodrugs offer great potential to overcome this challenge by enhancing drug penetration and tumor-killing efficacy. Squalene (SQ), a natural precursor for cholesterol biosynthesis with excellent biosafety and biocompatibility, can improve the membrane compatibility of hydrophilic drugs and enhance their cellular uptake when conjugated to chemotherapeutic agents or bioactive small molecules. In this study, we developed a novel lipophilic SQ-based prodrug system: the hydrophilic anticancer drug chidamide (CHI) was conjugated to SQ via an amide bond -- a linkage responsive to pancreatin and cathepsin B (key enzymes overexpressed in the PDAC microenvironment). This conjugation yielded an amphiphilic SQ-CHI prodrug, which was further self-assembled into folate (FA)-modified nanoparticles (FA-SQ-CHI NPs). The optimized NPs exhibited a uniform hydrodynamic diameter of 173.3 ± 1.5 nm, a polydispersity index (PDI) of 0.181 ± 0.18, a high drug loading capacity of 59.0% ± 0.77%, and a stable Zeta potential of -13.10 ± 0.86 mV. In vitro release studies showed that the NPs achieved 80.2% ± 4.22% cumulative drug release within 72 h in the presence of 0.25% pancreatin, while only 33%-38% release was observed in pH-adjusted buffers (pH 4.5 or 7.4) without enzymes. Cellular uptake assays confirmed that FA modification significantly enhanced intracellular delivery efficiency, with 1.8-2.3-fold higher fluorescence intensity in PDAC cells (PSN-1 and CFPAC-1) compared to non-targeted NPs at 12-24 h. The current protocol provides a comprehensive methodology for the synthesis and characterization of the prodrug, in vitro evaluation of enzyme-responsive release kinetics, and comparative analyses of therapeutic efficacy and tissue penetration, highlighting the nanocarrier's core advantages of targeted delivery, high drug loading, and enzyme-triggered controlled release.

Pancreatic neuroendocrine neoplasms (PanNENs) represent a rare and heterogeneous group of tumors with diverse biological behaviors and clinical outcomes, posing significant therapeutic challenges. Recent studies have suggested that certain proton pump inhibitors, including Lansoprazole, may possess direct anti-tumor properties beyond their classical role in acid suppression; however, their specific effects and molecular mechanisms in PanNENs remain largely unexplored. This study aims to investigate the anti-proliferative effects and elucidate the underlying molecular mechanisms of Lansoprazole in PanNEN models. Our findings demonstrate that Lansoprazole significantly upregulates the expression of DNA Damage Inducible Transcript 3 (DDIT3), a key stress-induced transcription factor. This upregulation leads to the subsequent inhibition of the oncogenic PI3K/AKT/mTOR signaling pathway, a central driver of cell growth and proliferation, resulting in marked suppression of PanNEN cell proliferation in vitro. Furthermore, we explored combination therapy strategies and found that Lansoprazole synergizes with everolimus, an established mTOR inhibitor used in PanNEN treatment. This combination enhances overall anti-tumor efficacy, suggesting a promising synergistic therapeutic strategy for PanNENs. These results not only reveal a novel, drug-repurposing approach for targeting PanNENs but also provide a mechanistic rationale for combining Lansoprazole with standard targeted therapies to improve patient outcomes.

Lung cancer has high mortality rates among both men and women worldwide. Nevertheless, mortality rates have been reported to decline with the advancement of novel therapeutic agents and the identification of new molecular targets. Schiff bases and triazine compounds have significant biological activity. For this purpose, new Schiff base derivative triazine compounds (TrzSchf 1-3) were synthesized in our study. The activities of the new compounds, characterized by spectroscopic techniques, against A549 lung cancer and MRC5 normal lung cells were identified in a series of studies. It was observed that TrzSchf 1-3 generally showed a growth-inhibitory effect against lung cancer cells (A549) and a non-toxic effect against normal lung cells (MRC5). Notably, TrzSchf 1 and TrzSchf 2, 3 exhibited prominent cytotoxic effects in A549 cells, with IC50 values of 14.24 and > 50 μM, respectively. It was observed that the compound with the most potent cytotoxicity against lung cancer cells was TrzSchf 1 (Selective Index: 3.62). In A549 cells, an increase in MAPK gene expression was observed for all compounds. It was observed that the expression of Caspase-3, CD40, CHK1, P27, P38, and P53 proapoptotic proteins increased by all compounds (TrzSchf 1-3), whereas the expression of antiapoptotic proteins such as BCL-2 and NFκB decreased by these compounds. The compounds are thought to be potential inhibitors of BCL-2 and NFκB, which are associated with cell death. Complementary and guiding in silico studies supported the experimental findings. BCL-2 was determined as the most favorable molecular target based on docking scores, and e-pharmacophore modeling further revealed key interaction features and enabled SAR analysis. The drug-likeness potential of the TrzSchf derivatives was evaluated based on Lipinski's Rule of Five parameters. Overall, both experimental and computational results suggest that TrzSchf 1-3 are promising lead candidates for further investigation in lung cancer therapy.

DHP107 is an oral paclitaxel enabling administration of paclitaxel without Cremophor EL, a vehicle used to improve the solubility of intravenous (IV) paclitaxel. The randomized phase II OPERA study investigated the efficacy and safety of DHP107 versus IV paclitaxel in patients with HER2-negative breast cancer.

Quantitative Structure-Activity Relationships are established in this work to predict the anticarcinogenic activities of flavonoids and related compounds in the MCF-7 breast cancer cell line. The selected descriptors tend to faithfully predict the [Formula: see text] and [Formula: see text] bioactivities, showing a straight-line trend in the correlation with the experimental data, even in this structurally diverse set of molecules. Finally, several structurally related compounds with unknown experimental anticarcinogenic activities are predicted. Therefore, the present study provides a guide for the rational design of potential new therapeutic molecules through the structure-activity parallelisms found. This work also demonstrates that QSAR models lead to evaluating whether the acquisition of experimental data is in agreement with the experimental protocol and methodologies, discriminating based on the domain of applicability and the fit to the QSAR model used.

Vinblastine (VBL) is a chemotherapeutic agent commonly used to treat malignant tumors in dogs. Although a maximum tolerated dose of 3.5 mg/m2 has been reported, the safety and tolerability of this dose in small dogs remain inadequately characterized.

Although FGFR2 is a well-validated oncogenic target, no selective FGFR2 inhibitors have been approved for clinical use. In this study, we report the discovery of 2H-pyrazolo[3,4-d]pyrimidin-4-amine derivative as novel, irreversible FGFR2 inhibitors. The optimal compound, PLW559, potently inhibited FGFR2 with an IC50 value of 13.59 nM and demonstrated exceptional selectivity over FGFR1, FGFR3, and FGFR4. Covalent binding to the target was confirmed by mass spectrometry. In cellular models, PLW559 exhibited potent and selective antiproliferative effects against FGFR2-driven cancer cells, effectively suppressed downstream FGFR2 signalling and induced cancer cell apoptosis. Notably, it showed minimal activity in non-FGFR2-dependent cells. This work presents a new class of selective FGFR2 inhibitors based on a novel scaffold, offering promising lead compounds for the development of FGFR2-target therapies.