Abstract Background Mesothelioma (Me) is an aggressive cancer with limited response to conventional therapies. The tumor’s harsh microenvironment contributes to immune escape and therapy resistance and the effects of ICIs on Me are still unclear. Adenosine, an immunosuppressive molecule produced from AMP by the enzyme CD73, accumulates in hypoxic tumor areas. Elevated CD73 and adenosine receptor A2B (A2Br) levels on Me cells are linked to worse patient outcomes, indicating their important role in disease progression and potential as targets for treatment. Aim This study characterizes the Me-ME (micro environment) and evaluates the efficacy of TT-4 (A2B inibitor) and AB680 (CD73 inibitor), alone or with aPD-1, using 3D models in vitro and in vivo . Methods CD73 and A2B receptor levels were quantified in tumor and normal samples using qRT-PCR and IHC. Cells lines were treated with CoCl 2 to mimic hypoxia, then CD73, A2Br and related markers were analyzed. MSTO-211H and REN cells were silenced for CD73, grown as spheroids and adenosine release was measured. Co-culture spheroids of MSTO-211H and Jurkat cells were treated with AMP and CD73 inhibitor, then analyzed for viability and immune markers. An orthotopic Me model was established by injecting AB1-B/c-LUC cells and monitored by in vivo imaging. Proteomic analysis of spheroids was conducted to identify proteins and pathways involved. Results Hypoxia boosts CD73 and A2Br expression in Me cells, leading to adenosine production via CD73. In 3D co-cultures, AB680 lowered Me cell viability and enhanced activation of Jurkat T cells. In mice, combining aPD-1 therapy with A2Br or CD73 inhibitors strongly reduced tumor growth. Proteomics identified 93 proteins influenced by adenosine signaling through A2B. Conclusion Targeting the adenosine pathway alongside PD-1 blockade offers a promising new immunotherapy strategy for Me.

AimTo investigate the expression of succinate dehydrogenase B and its potential as a therapeutic target in pleural mesothelioma (PM).MethodsThe expression of succinate dehydrogenase B was examined in cell lines using PCR. The expression of succinate dehydrogenase B mRNA was subsequently examined in a series of fresh-frozen primary patient samples using reverse transcription polymerase chain reaction. The expression of succinate dehydrogenase B protein was examined by immunohistochemistry on a pleural mesothelioma tissue microarray derived from Formalin-Fixed and Paraffin-Embedded specimens to determine if succinate dehydrogenase B expression correlated with survival benefit. Using a resazurin-based assay we examined whether targeting the mitochondrial complex II (containing succinate dehydrogenase B) with a chemical inhibitor could have effects on cell proliferation.ResultsTotal ribonucleic acid was isolated from a panel of mesothelial-derived cell lines (both cancerous and non-cancerous), and expression of succinate dehydrogenase B messenger RNA was assessed by RT-PCR and found to be ubiquitously expressed. Total Ribonucleic acid was isolated from a panel of fresh-frozen surgical specimens, and succinate dehydrogenase B was significantly overexpressed in mesothelioma at the messenger RNA level. Immunohistochemical staining and analysis of a mesothelioma tissue microarray showed that expression did not correlate with any survival benefit, confirmed by in silico analyses of other datasets. However, links between DNA methylation at individual residues of the succinate dehydrogenase B gene and overall survival were identified, along with correlations to immune cell infiltrates. Targeting the mitochondrial complex II with oxaloacetic acid did not show any significant potential for higher efficacy in cell lines derived from malignant tumors compared to those derived from normal mesothelial cells.ConclusionsOur results demonstrate that succinate dehydrogenase B is overexpressed in PM but may not be a suitable candidate for therapeutic targeting in this disease.

Mesothelioma is a rare and aggressive cancer with a poor prognosis and limited therapeutic options. Despite recent advances, conventional treatment approaches remain largely ineffective due to late diagnosis, chemoresistance and immunosuppressive tumor microenvironment. This review reports the latest studies on combination therapies for mesothelioma, focusing on the potential of integrating chemotherapeutic agents, molecularly targeted agents, vaccines and natural bioactive compounds such as polyphenols. Clinical and preclinical studies demonstrate that integrating immune-modulating drugs or molecular inhibitors with chemotherapy can improve survival and reduce tumor progression in mesothelioma models and patients. Vaccine-based strategies show potential for inducing host-persistent immune responses when combined with conventional treatments. Moreover, natural compounds such as polyphenols show synergistic effects with chemotherapeutics and targeted agents by modulating several signaling pathways involved in cancer cell growth and progression and by overcoming drug resistance. While several combination strategies are under clinical investigation, further studies are needed to develop more effective and personalized therapeutic approaches that could be translated into standardized treatment protocols.

Cancer disparities in low- and middle-income countries (LMICs) persist because of socioeconomic inequalities and limited access to screening infrastructure, which requires equitable diagnostic solutions. As researchers, we need to develop interventions which mirror successful strategies from high-income countries (HICs) to address mortality inequalities. Routine cancer diagnosis functions as a fundamental element of effective management yet remains unavailable to numerous populations in LMICs. This review proposes the conceptual “OncoCheck” model, which combines the terms Oncology “Onco” and Screening “Check” as an integrated approach to early cancer detection. It provides a theoretically sound practical approach that combines liquid biopsy with point-of-care testing (POCT) and artificial intelligence (AI) to achieve high-sensitivity diagnostics in resource-limited settings without requiring advanced infrastructure. The review advocates OncoCheck as a promising and practical cancer screening solution which shows potential to increase accessibility and decrease costs while improving survival rates through early detection. Moving beyond technical specifications, the manuscript assesses its socioeconomic impact, showing reduced medical costs and improved treatment outcomes. The paper describes its implementation framework together with a validation strategy and performance benchmarks. The analysis further focuses on the implementation barriers like algorithmic bias mitigation, infrastructure limitations, and ethical AI deployment. The OncoCheck system delivers equitable cancer care by implementing a hospital-at-home model which functions with real-world health systems.

[This corrects the article DOI: 10.3389/fonc.2022.895708.].

Background Immune checkpoint inhibitors have shown inconsistent benefit in second-line malignant pleural mesothelioma. Although histology and several molecular biomarkers, such as BAP1 and others, reveal substantial heterogeneity, their contribution to treatment selection remains uncertain. Because informative censoring may bias time-to-event analyses, we re-evaluated key second-line trials to determine whether censoring patterns contributed to discordant efficacy results. Methods Published Kaplan–Meier curves from CONFIRM (nivolumab vs placebo), the CONFIRM update, and PROMISE-Meso (pembrolizumab vs chemotherapy) were reconstructed. Censoring distributions were quantified using reverse-KM, and differential censoring between arms and across endpoints (overall survival [OS] vs progression-free survival [PFS]) was evaluated over time. Sensitivity analyses were applied to address excessive censoring, and hazard ratios and restricted mean survival time differences were calculated. Results In CONFIRM, the initially reported OS advantage coincided with short follow-up and a differential censoring pattern consistent with post-progression censoring. The updated CONFIRM dataset, with longer follow-up, demonstrated no OS benefit and no evidence of post-progression censoring. In contrast, PROMISE-Meso showed no evidence of post-progression censoring at any time and remained negative throughout. When general differential censoring was addressed using sensitivity analyses, both the PFS effect and the OS advantage in CONFIRM lost statistical significance. Conclusions Post-progression censoring is an under-recognized source of bias that can generate misleading survival signals and may have contributed to the initial CONFIRM findings that were based on a premature early data lock. Although a modest treatment effect in a biologically defined subgroup cannot be excluded, current evidence remains inconclusive. Future studies should ensure comprehensive ascertainment of survival outcomes and include adequately powered biomarker programs, including histology and BAP1 status.

Although immune-checkpoint inhibitors (ICIs) have positively impacted the treatment of some tumors, several evidences clearly suggest that the actual relevance of these treatments has not been fully unraveled yet Hypoxic tumors activate the purinergic pathway, which acts as a major immunosuppressive mechanism. CD39 and CD73 are enzymes involved into ATP degradation to AMP and AMP degradation to the immune-suppressive adenosine, respectively thus the inhibitors of these pathways are progressively attracting the attention as actionable targets for immunotherapy in many solid tumors MPM is a highly aggressive malignancy whose treatment challenging for different reasons, including among others acidic and hypoxic tumor microenvironment (TME). Hypoxic TME and the expression of purinergic receptors and enzymes on tumor cells are considered criteria to select patients for immunotherapy with adenosine pathway inhibitor. We have already shown the expression of adenosine receptor 2 B (ADORA2b) and CD73 enzyme on MMP cells is inversely related to patients' survival and that MMP cells degrade AMP to adenosine via CD73. Aim of our study was assessing if ADORA2B and or CD73 with or without PD-1 ICi were able to curb tumor cells growth in a syngeneic mouse MPM model. For orthotopic mouse models of MPM, CD73 ADORA2B positive 105 AB1B-LUC cells were injected intraperitoneally (i.p.) in immunocompetent BALB/C mice. After three weeks we started monitoring tumor growth and the animals were randomized into five groups of three mice each for treatment with PBS 100 μl (control), Pembro (anti murine PD-1; MCE ) 4.4 mg/Kg, TT-4 (ADORA2 Inhibitor Portage), 3 mg/Kg, Pembro+TT-4, AB-680 (CD73 inhibitor, MCE, HY-125286) 20 mg kg) and Pembro+AB-680 were injected every two days for three weeks. Disease was measured with IVIS® Lumina XR in vivo imaging system, thus facilitating non-invasive longitudinal monitoring of disease progression or regression. after 21 days AB680 and TT4 resulted to be the most effective single treatments (fluorescence reduction vs control p<0.01) whereas Pembro+TT4 Pembro+AB680 did not differ significantly from the single treatments. IHC of murine samples from mice treated as above revealed infiltration of CD8 T cells mostly with AB680 single agent or in combo compared to the cold pattern of the control samples. No relevant toxicity was recorded. Inhibition of the purinergic pathway exerts a significant immune-mediated MPM growth inhibition luciano Mutti, Antonio Giordano, Pierpaolo Correale, Caterina Costa. Adenosine production suppression and PD-1 immune check point inhibitor for treatment of malignant pleural mesothelioma. (MPM) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 2247.

PURPOSE:No guidelines exist to delineate radiation therapy (RT) targets for the treatment of multiple glioblastoma (mGBM). This study analyzes margins around the gross tumor volume (GTV) to create a clinical target volume (CTV), comparing response parameters and modalities of recurrence. Material and Methods: One-hundred and three mGBM patients with a CTV margin of 2 cm (GTV + 2.0 cm) or 1 cm (GTV + 1.0 cm) were retrospectively analyzed. All patients received a total dose of 59.4-60 Gy in 1.8-2.0 Gy daily fractions, delivered from 4 to 8 weeks after surgery, concomitantly with temozolomide (75 mg/m2). Overall survival (OS) and progression-free survival (PFS) were calculated from the date of surgery until diagnosis of disease progression performed by magnetic resonance imaging and classified as marginal, in-field, or distant, comparing site of progression with dose distribution in RT plan. RESULTS:OS in mGBM CTV1 group was 11.2 months (95% confidence interval [CI], 10.3-12.1), and 9.2 months in mGBM CTV2 group (95% CI, 9.0-11.3). PFS in mGBM CTV1 group occurred within 8.3 months (95% CI, 7.3-9.3), and 7.3 months in mGBM CTV2 group (95% CI, 6.4-8.1). No difference was observed between the two groups in terms of OS and PFS time distribution. Adjusted to a multivariate Cox risk model, epidermal growth factor receptor amplification resulted a negative prognostic factor for both OS and PFS. CONCLUSION:In mGBM, the use of a 1 cm CTV expansion seems feasible as it does not significantly affect oncological outcomes and progression outcome.

Immune-checkpoint inhibitors (ICIs) were proven effective in inducing tumor regression. However, its toxicity tends to be fatal. We sought to investigate the hospital volume/outcomes relationship. Databases were searched for studies reporting immune-checkpoint inhibitors adverse events (AEs) in patients with solid-organ malignancies. The outcomes were A) the pooled events rate (PER) of grade 5, grade 3-4, cardiac-related, and pulmonary-related AEs, and B) the assessment of the volume/outcomes relationship. One hundred and forty-seven studies met our inclusion criteria. The PER of grade 5, grade 3-4, and any-grade AEs was 2.75% (95%CI: 2.18-3.47), 26.69% (95%CI: 21.60-32.48), and 77.80% (95%CI: 70.91-83.44), respectively. The PER of pulmonary-related AEs was 4.56% (95%CI: 3.76-5.53). A higher number of annual cases per center was significantly associated with reduced grade 5 (p = 0.019), grade 3-4 (p = 0.004), and cardiac-related AEs (p = 0.035) in the meta-regression. In the current era of cancer immunotherapy, knowledge regarding the early diagnosis and management of immunotherapy-related AEs is essential. Our meta-analysis demonstrates the importance of center volume in improving outcomes and reducing the incidence of severe AEs.

Abstract Background The programmed death ligand 1 (PD-L1) has recently emerged as a target immunotherapy in Triple-negative breast cancer (TNBC). However, the tumor-intrinsic role of PD-L1 and its pathway still needs to be fully clarified. Recently, a close association between CD73, PD-L1, cancer cell steaminess and epithelial-to-mesenchymal transition phenotypes is emerging. In addition, we have previously demonstrated that miR-320a and miR-145 target PD-L1, whereas miR-30 family is known to target CD73. In this study, we aimed at investigating the role of these miRNAs as prognostic and predictive dynamic biomarkers for immunotherapy in TNBC. Additionally, we investigated whether PD-L1 exerts cellular autonomic functions in TNBC beyond its role in the immune checkpoint. Results CD73 and PD-L1 expression was assessed in a panel of breast cancer lines and the non-malignant breast cells MCF-10. We showed a significantly higher CD73 and PD-L1 expression in MDA-MB-231 (TNBC, Basal B) than in MCF-10. Conversely, in MCF-7 cells (HR +, Luminal A), CD73 and PD-L1 expression is lower than in MCF-10. Furthermore, we showed that low PDL1 expression is correlated with high levels of miR-320a and miR-145, and CD73 expression is inversely related to miR-30 expression. These putative biomarkers were also validated in two patients with different PD-L1 status. From the analysis of the data obtained by real-time qRT-PCR, we found high plasma level of CD73 and low plasma level of miR-320a, miR-145 and miR-30 in the patient with PD-L1 positive BC. On contrary, in the patient with PD-L1-negative BC, we found low plasma level of CD73 and high plasma level of miR-320a, miR-145 and miR-30. Furthermore, to explore whether PD-L1 could also have an intrinsic role in tumor development and invasiveness, we used stably MDA–MB-231 PD-L1-silenced cells. We found that cell growth, colony formation, migration rate and the ability to form spheres are consistently reduced upon shRNA-mediated PDL1 silencing. In particular, we observed a reduction in the number, diameter and volume of the 3D spheres compared to the control cells. To further demonstrate the role of PD-L1, we treated the PDL1-high expression cells MDA-MB-231, PD-L1 silenced MDA-MB-231 clones, and PD-L1 low expression cells MCF-7 with Durvalumab, an anti-PD-L1. Interestingly, we observed that in PD-L1 silenced clones and MCF-7 cells, the sphere-forming ability was increased in presence of Durvalumab treatment. On the contrary, in MDA-MB-231, Durvalumab inhibited the sphere-forming ability. Conclusion Our data confirm that PD-L1 and CD73 are targets of miR-320a/miR-145 and miR-30 respectively. These could be potential predictive dynamic biomarkers for chemotherapy, ICIs and anti-CD73 therapeutic approach. Future validations of these biomarkers in an extensive series of TNBC patients are needed to support their use in clinical practice. Here, we further characterized the cellular autonomic role of PD-L1 in breast cancer and showed a differential role of basal PD-L1 expression in PD-L1 checkpoint inhibitors treatment efficacy. This suggests a potential role in monitoring PD-L1 expression indirect biomarkers (i.e. miR-320a, miR-145 and CD73) during ICIs treatment. References 1. Costa C, Indovina P, Mattioli E, Forte IM, Iannuzzi CA, Luzzi L, Bellan C, De Summa S, Bucci E, Di Marzo D, De Feo M, Mutti L, Pentimalli F, Giordano A. P53-regulated miR-320a targets PDL1 and is downregulated in malignant mesothelioma. Cell Death Dis. 2020 Sep 14;11(9):748. doi: 10.1038/s41419-020-02940-w. PMID: 32929059. Citation Format: Caterina Costa, Claudia von Arx, Alessandra Calabrese, Francesca Pentimalli, Mutti Luciano, Antonio Giordano, Michelino De Laurentiis. Identifying new immune-related biomarkers in TNBC with a look at PD-L1 cell-autonomous role [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-15-10.