Immunotherapy has introduced a new era in cancer treatment. The clinical goal of cancer immunotherapy is to prime the host immune system to provide passive or active immunity against malignant tumors. We have previously demonstrated that Brucella abortus (Ba) RNA downregulates IFN-γ-induced MHC-I surface expression in human monocytes/macrophages via a TLR8-dependent mechanism. Other bacterial RNAs can mimic this phenomenon. The presence and activity of NK cells in tumors have been correlated with better patient survival, supporting the evidence that these cells are essential in the immune response against tumors. So, we postulated that bacterial RNA (bacRNA) can be used to modulate MHC-I expression in tumors to enhance the NK cell response. Hence, the aim of this study was to investigate the immunomodulatory role of bacRNA in solid tumors. We first stimulated human glioblastomas U251 and LN-229, colorectal adenocarcinoma HT-29, breast cancer MCF-7, and murine melanoma B16-OVA cells with bacRNA in the presence of IFN-γ. Our experiments demonstrated that bacRNA diminished IFN-γ-induced MHC-I surface expression in all tumor cell lines. Moreover, the hTLR8 agonist ORN06/LyoVec mimicked the effect of bacRNA, indicating that MHC-I reduction would be mediated by TLR8. In addition, the decrease in MHC-I mediated by bacRNA correlated with increased NK cytotoxicity. Finally, treatment with either bacRNA or the ORN06 agonist resulted in greater immune cell infiltration and activation within the tumor compared to untreated mice. Furthermore, bacRNA delayed tumor growth in the B16 melanoma model. Overall, our established model of MHC-I downregulation (either by bacRNA or synthetic hTLR8 agonists) could be used as a therapeutic strategy to promote anti-tumor responses.

Kaempferol exhibits promising anti-colorectal cancer (CRC) activity, but its intracellular exposure and efficacy are strongly influenced by phase II metabolism and efflux transport. Here, we show that UGT1A1 and UGT1A9 differentially regulate kaempferol accumulation and anti-CRC activity, and that ABCG2/BCRP and Mdr1/P-gP play distinct roles in its intracellular retention and pharmacological effects. Anti-CRC activity was evaluated using sulforhodamine B, colony formation, wound-healing, Transwell, apoptosis, and xenograft assays. Knockdown models of UGT1A1/UGT1A9 and BCRP/P-gP were combined with laser scanning confocal microscopy and LC/MS to characterize intracellular exposure, metabolite disposition, and functional consequences. SIP-based proteomics, bioinformatic analyses, molecular docking, and orthogonal validation assays were used to prioritize and validate a nuclear target of kaempferol. Kaempferol significantly suppressed tumour growth and inhibited CRC cell proliferation and migration. UGT1A1 knockdown increased intracellular accumulation of kaempferol, including nuclear exposure, but reduced cytotoxicity, whereas UGT1A9 knockdown enhanced cellular sensitivity. Similarly, BCRP knockdown increased intracellular accumulation and anti-CRC efficacy, whereas Mdr1 knockdown attenuated the antitumour effect despite increasing total intracellular accumulation. Mechanistically, these differential effects were associated with altered metabolite composition, particularly the balance between the active metabolite K-7-G and the weakly active metabolite K-3-G. BRG1 was prioritized as a functionally relevant nuclear target of kaempferol, and kaempferol promoted its ubiquitination-dependent proteasomal degradation. Together, these findings show that kaempferol efficacy is determined not simply by intracellular accumulation, but by the coordinated interplay among phase II metabolism, efflux transport, metabolite disposition, and nuclear target engagement. This study provides a mechanistic basis for improving kaempferol-based CRC therapy.

Cancer-associated fibroblasts (CAFs) are key components of the melanoma tumor microenvironment and have been implicated in immune evasion and therapy resistance. However, the spatial relationships between CAFs, extracellular matrix (ECM) remodeling, and immune exclusion remain incompletely characterized.

We integrated single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics from primary melanoma samples (892 regions of interest) to investigate the landscape of CAF-immune interactions. CAF scores were calculated based on five fibroblast-associated genes. Differential expression analysis, Spearman correlation, and cell state subclustering were performed to characterize the spatial organization of the tumor microenvironment. Findings were examined using TCGA bulk RNA-seq data and drug sensitivity was assessed using the GDSC database.

CAF-high regions exhibited significantly lower immune scores compared to CAF-low regions (ρ = -0.442, p < 0.001), with enrichment of ECM-related genes (COL1A1, COL1A2, DCN, FBLN1) and downregulation of immune genes (PTPRC, CD4, CD8A). Spatial co-expression of FN1 and its integrin receptor subunit ITGB1 was observed (ρ = 0.100, p = 0.003), supporting a potential role for ECM-receptor signaling in immune exclusion. Six distinct melanoma cell states were identified, with CAF-associated tumor states (State 4) exhibiting significantly higher proliferative capacity than immune-associated states (State 5) (p = 0.007). The CAF signature correlated with cisplatin sensitivity (ρ = -0.627, p < 0.001) and trametinib resistance (ρ = 0.354, p = 0.009).

This study provides a spatial atlas of CAF-immune interactions in melanoma. CAF abundance is negatively correlated with immune infiltration at the spatial level, with ECM remodeling and FN1-ITGB1 signaling potentially mediating this immune exclusion. The CAF signature showed correlations with drug sensitivity in cell line models, warranting further investigation in CAF-containing systems.

To investigate the anticancer effects of asiatic acid (AA) and asiaticoside (AS) in cisplatin (Cis)-resistant neuroblastoma SH-SY5Y and neural stem cells derived from human stem cells from apical papilla (NSCs-hSCAPs).

An MTT cell viability assay was performed to determine Cis toxicity and assess the cytotoxic effect of AA or AS on Cis-treated SH-SY5Y and the effect on NSCs-hSCAPs. Subsequently, the combination index (CI) was calculated to observe occurring interactions between the compound and Cis. For mechanistic exploration, flow cytometry using annexin V and PI staining and a caspase-3 activity assay were done to quantify apoptotic cell death. qRT-PCR was used to measure apoptotic and antioxidant mRNA levels. Additionally, the protein levels were analyzed by using the Western blot technique. Finally, to illustrate the putative interactions between the compound and its target, molecular docking was trialled.

MTT results revealed that AS did not affect both cells. However, 20 µM AA in combination with 2.5 µM Cis could significantly reduce SH-SY5Y cell viability with a CI value of 0.41, indicating their synergistic effect. Moreover, the effective concentration of AA did not affect Cis-treated NSCs-hSCAPs, suggesting its low toxicity. Considering SH-SY5Y, AA alone or its combination dramatically increased % apoptotic cells, which showed a minimal increment in Cis alone. Importantly, the combination treatment significantly increased caspase-3 activity. Mechanistically, BAX levels were elevated, whereas BCL2, SOD-1, and HO-1 levels were downregulated by AA alone or in combination. These results were also consistent with Western blot analysis. Interestingly, an inhibition of GSK3β was observed in the combination treatment, suggesting that these outcomes may be related.The molecular docking illustrated that AA interacted with GSK3β and BCL-2 through multiple residues, indicating their potential binding interactions.

AA and Cis treatment suppressed SH-SY5Y cell viability by enhancing apoptosis, reducing antioxidant gene expression, and potentially inhibiting GSK3β.

VISTA (V-domain Ig suppressor of T-cell activation; gene symbol VSIR) has emerged as a distinct immune checkpoint with increasing relevance in cancer biology and immunotherapy. Unlike classical checkpoint pathways that are predominantly interpreted through T cell-centered suppression, accumulating evidence indicates that VISTA exerts broader and highly context-dependent immunoregulatory functions, frequently centered on myeloid cells within the tumor microenvironment. In many malignancies, VISTA is preferentially enriched in tumor-associated macrophages, myeloid-derived suppressor cells, and dendritic cells rather than being uniformly expressed by tumor cells. This compartment-specific distribution has been linked to suppressive myeloid differentiation, impaired antigen presentation, altered inflammatory cytokine programs, T-cell dysfunction, and immune exclusion. Recent studies further suggest that VISTA biology is shaped by ligand context, tissue compartment, tumor acidity, and therapy-induced adaptation, supporting its role in resistance to conventional immune checkpoint blockade in selected settings. In this review, we summarize the structural and functional features of VISTA, its expression patterns in physiological and tumor contexts, its mechanistic role in shaping the tumor microenvironment, and the current landscape of VISTA-targeted therapeutic strategies. Key unresolved questions include which VISTA binding partners dominate in vivo, how cellular source and spatial localization determine biological and prognostic meaning, and whether early pharmacodynamic target engagement will translate into durable clinical benefit. Collectively, current evidence supports the view that VISTA should be considered not simply as another inhibitory receptor, but as a dynamically regulated, myeloid-centered checkpoint axis with translational relevance in checkpoint-refractory and immune-excluded tumors. A compartment-resolved understanding of VISTA expression and function may improve prognostic interpretation, biomarker development, and patient selection for VISTA-directed therapies.

Head and neck squamous cell carcinoma (HNSCC) is a common malignancy characterized by poor survival due to recurrence, metastasis and therapy resistance. In addition to genetic alterations, metabolic reprogramming is a hallmark of HNSCC and contributes to tumor progression and treatment failure. The hepatocyte growth factor (HGF)/c-MET signaling pathway is frequently activated in head and neck squamous cell carcinoma (HNSCC), where it promotes tumor cell proliferation, invasion, and increased glucose metabolism. However, its contribution to the regulation of glutamine metabolism in HNSCC remains largely unexplored. Basal GLS-1 expression was compared between primary human oral keratinocytes (HOK) and HNSCC cell lines with distinct MET status (Detroit 562, FaDu, and SCC-154). Subsequently, HNSCC cell lines were stimulated with HGF, and GLS-1 expression was assessed by quantitative PCR and Western blotting. Functional effects were evaluated using wound-healing assays, enzymatic quantification of extracellular glutamine and glutamate, and Seahorse-based mitochondrial respiration analysis. GLS-1 was silenced using siRNA to determine its functional relevance. In addition, transcriptomic data from the TCGA-HNSC cohort were analyzed to evaluate the clinical association between MET signaling, MAPK/ERK pathway activity, and GLS1 expression. HGF stimulation selectively induced GLS-1 expression in MET-amplified Detroit 562 cells at both mRNA and protein levels, whereas MET wild-type cells remained largely unaffected. Mechanistically, HGF triggered robust ERK1/2 activation, and pharmacological inhibition of c-MET or ERK signaling abrogated GLS-1 induction. Functionally, HGF increased glutamine consumption, glutamate production, and mitochondrial respiratory capacity in a GLS-1-dependent manner, while GLS-1 silencing significantly impaired HGF-induced cell migration and mitochondrial respiration. Importantly, analysis of TCGA-HNSC tumors revealed significant correlations between MET expression, MAPK/ERK pathway activity, and GLS1 expression, supporting the clinical relevance of the identified signaling axis. Together, these findings identify GLS-1 as a downstream effector of HGF/c-MET-MAPK/ERK signaling in MET-amplified HNSCC and link oncogenic signaling to glutamine metabolism and mitochondrial function. The observed associations in TCGA patient tumors further support the clinical relevance of this pathway and suggest that targeting glutaminase or ERK signaling may represent a promising therapeutic strategy in c-MET-driven HNSCC.

Problems of low bioavailability, rapid systemic degradation, non-selective targeting, and high toxicity often mar conventional cancer therapies. Cordycepin (COR), a natural adenosine analogue with potent anticancer activity, particularly against breast cancer, is limited by its high rate of inactivation through the enzyme adenosine deaminase, instability at stomach pH, and poor pharmacokinetics, thus reducing its therapeutic potential. To address these limitations, we have designed a solid lipid nanoparticles (SLNs)-based delivery system for the effective entrapment and release of COR to improve its stability, cellular uptake, and cytotoxic efficacy against MCF-7 human breast cancer cells. The synthesized COR-loaded SLNs (CSLN) were found to be monodispersed, spherical nanoparticles with good physical and chemical stability. In vitro release studies exhibited sustained release behaviour in acidic and physiological conditions, mimicking the endo-lysosomal and systemic circulation pH, respectively. The results of cytotoxicity assays indicated that CSLN significantly enhanced the cytotoxic efficacy of COR against MCF-7 cells as contrasted to the free drug. Improved cellular uptake and greater drug accumulation inside cells led to improved apoptotic activity and thus, more clonogenic activity. These findings suggest that SLNs-based nanoencapsulation is a promising approach for enhancing the therapeutic efficacy and safety of COR for breast cancer therapy.

Glioblastoma (GBM) is extremely difficult to treat due to the poor blood-brain barrier (BBB) penetration, strong extracellular matrix, high interstitial pressure, and restricted cellular absorption, all of which prevent efficient drug delivery. Folate-conjugated nanotheranostics have emerged as a viable way to overcome these hurdles, taking advantage of GBM cells' upregulation of folate receptors (FRs). Folate-conjugation chemistry, either by direct ligand coupling or linker-assisted surface modification, have resulted in more selective receptor-based targeting, increased stability, and regulated chemotherapeutic release inside the tumor microenvironment. These nanosystems show increased penetration and selective absorption via FR-mediated endocytosis, which allows for deeper intratumoral distribution and more effective administration of chemotherapeutics and imaging agents. Recent advances in nanomedicine-based drug delivery systems with folate-functionalization shows promise for integrated GBM diagnosis and treatment. Latest findings also suggest good biocompatibility and low systemic toxicity of folate-armed nanotheranostics; however long-term safety is still a crucial issue and need to be discussed. This review summarizes the key challenges to GBM therapy and explores how folate-decorated nanotheranostics, via tailored chemistry and increased tumoral contact, constitute a potential option for GBM precision therapy.

Colorectal cancer (CRC) remains a major cause of cancer-related mortality worldwide, and patients with advanced or metastatic disease still require more effective targeted therapeutic options. N-acetyltransferase 10 (NAT10), an RNA acetyltransferase responsible for N4-acetylcytidine (ac4C) modification, has been implicated in CRC progression, metastasis, immune evasion, and therapy resistance. The recently resolved human NAT10 structure (PDB ID: 9J3C) provides a structural basis for rational discovery of NAT10-directed chemical scaffolds. We combined large-scale virtual screening, molecular simulation, and cellular testing to identify NAT10-targeting compounds. Approximately 300,000 molecules were screened by hierarchical docking, followed by ADMET prediction, 100-ns molecular dynamics simulations, and MM/PBSA analysis. G856-6814 showed a predicted binding free energy of - 26.33 ± 3.37 kcal/mol and inhibited HCT116 cell viability with an IC₅₀ of 211.6 ± 14.8 nM. In a recombinant enzyme assay, G856-6814 inhibited NAT10 acetyltransferase activity with an IC₅₀ of 2.3 ± 0.4 µM. G856-6814 treatment increased the apparent NAT10 melting temperature from 50.4 ± 0.5 to 56.0 ± 0.8 °C. Treatment also reduced global RNA ac4C and ac4C enrichment on c-MYC, CCNA2, and CCNB1 transcripts. G856-6814 was 19.6-fold less potent against TIP60 and more than 43-fold less potent against p300 and GCN5. Together with the attenuation of cellular effects after NAT10 knockdown, these findings support G856-6814 as a NAT10-targeting inhibitory scaffold that requires broader selectivity and in vivo evaluation.

Cutaneous melanoma (CM) is the most aggressive form of skin cancer, characterized by high metastatic potential and resistance to conventional therapies. Evidence suggests that modulation of purinergic signaling, interleukin-6 (IL-6), and NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) may represent promising strategies for treating CM. Calcitriol, the active form of vitamin D, has demonstrated antineoplastic potential across different cancer types. In addition, reports have shown that purinergic signaling can be modulated by calcitriol. However, its mechanism of action on the purinergic system in melanoma remains poorly explored. In this study, we investigated the antineoplastic effects of calcitriol on human cutaneous malignant melanoma cell lines, with a focus on the purinergic system. The cells A375 and SK-MEL-28 were treated with calcitriol at 1, 10, and 50 nM for 24 h, and we analyzed cell viability, mitochondrial membrane potential, apoptotic body detection, migration, and ectonucleotidase enzymatic activity. We also analyzed the expression of ectonucleotidases (CD39 and CD73) and IL-6/NLRP3. We found that calcitriol significantly reduced A375 cell viability, altered mitochondrial potential, inhibited cell migration, and induced the formation of apoptotic bodies. These effects were not observed in SK-MEL-28. Moreover, calcitriol modulated the ectonucleotidases activity and downregulated CD39 and CD73 expression. Additionally, the treatments downregulated IL-6 and NLRP3 expression. Thus, our findings demonstrate that calcitriol exerts antineoplastic effects on melanoma cells via mitochondrial dysfunction, apoptosis-related alterations, inhibition of migration, and modulation of purinergic and inflammatory mediators. Therefore, calcitriol is a promising adjuvant for CM therapy.