Breast Cancer (BrCa) remains a devastating disease presenting emerging needs for effective management. Recently, epigenetic biomarkers are assessed in liquid biopsy for diagnostic and prognostic applications. This study applies a 3-step data-driven biomarker discovery pipeline to identify robust methylation biomarkers and generate high-performance biosignatures specific for clinically significant BrCa end-points, followed by laboratory validation in patient cell-free DNA (cfDNA). Publicly available genome-wide methylomes from 520 BrCa and 185 non-diseased breast tissues (discovery dataset) were analyzed via Automated Machine Learning (AutoML, JADBio) to identify BrCa-specifically methylated promoters. Bioinformatic search revealed any BrCa biological relevance. Next, the methylation of identified promoters was experimentally validated in plasma cfDNA from 195 BrCa patients and 135 healthy individuals by Methylation Specific qPCR (qMSP) (validation cohort). Finally, autoML analyzed experimental and clinical data to develop optimized classifying biosignatures for diagnosis, prognosis, and prediction. AutoML identified 3 BrCa-specific methylated promoters in CLDN15, MRGPRD and ZNF430. Pathway analysis revealed implications with biological processes such as signaling and transcription. Laboratory validation using clinical cfDNA samples confirmed elevated methylation levels in BrCa patients for all 3 promoters, which were correlated with poor prognostic and predictive parameters. Classification analysis by autoML of experimental methylation measurements and patients’ clinical data built 5 specific models: a diagnostic biosignature distinguishing BrCa from health (AUC 0.79, CI: 0.75–0.84), a classification biosignature differentiating BrCa disease status (adjuvant, neoadjuvant, and metastatic group) (AUC 0.68, CI: 0.62–0.72), a prognostic biosignature predicting relapse (AUC 0.79, CI: 0.74–0.83), a biosignature predicting treatment response in metastatic patients (AUC 0.86, CI: 0.67–1.00), and a biosignature differentiating distinct molecular subtypes (AUC of 0.71, CI: 0.64–0.77), underscoring their possible clinical utility. Our data-driven approach successfully identified 3 BrCa-specifically methylated promoters in genes not previously implicated in BrCa. Their role in pathology needs further attention as they could also represent novel targets. Moreover, the laboratory validation in clinical BrCa cfDNA samples led to the development of 5 biosignatures, some demonstrating strong predictive performance. The low number of features and the minimally invasive nature of liquid biopsy highlight the potential for clinical implementation of great value.

In view of the shifting standard of care for non-small cell lung cancer (NSCLC), we aimed to capture the evolution of the epidemiological and systemic treatment (ST) landscape in Greece. This non-interventional, single-country, multicenter, retrospective study, collected high-level aggregate data for adult patients who were newly diagnosed with histologically/cytologically confirmed Stage I-IV NSCLC between 01-Jan-2016 and 31-Dec-2020 (index period) at leading oncology clinics across the country. Overall, 10,644 patients were newly diagnosed with NSCLC during the entire 5-year index period at the 18 participating sites (63.8

e13064 Background: The detection and enumeration of circulating tumor cells (CTCs) in the peripheral blood is an independent poor prognostic factor for patients (pts) with metastatic breast cancer (MBC), while longitudinal CTC assessment holds promising role also as a real-time biomarker for monitoring treatment efficacy and disease progression. CDK4/6 inhibitors represent a key treatment option for hormone receptor-positive (HR+) MBC. We herein aimed to assess the clinical value of CTCs and their kinetics for HR+ MBC pts treated with CDK4/6 inhibitors. Methods: Peripheral blood (PB) was obtained from pts with HR+ MBC: a) at baseline before the start of CDK4/6 inhibitors plus endocrine treatment (N = 39), and b) at the first evaluation of treatment response (N = 24/39 pts; paired samples). CTCs were in parallel isolated using Ficoll density gradient centrifugation, and the automated size-based Parsortix system (ANGLE plc). CTC detection and enumeration was performed by immunofluorescence staining for cytokeratins (CKs)/CD45/dapi and observation via fluorescence microscopy. Results: CTCs (CK+/CD45- cells) were detected by any assay in 15/39 (38.5%) of pts at baseline and in 5/24 (20.8%) at the first evaluation, with a total of 71 and 25 CTCs identified, respectively (mean CTC No per patient: n = 1.82 and 1.04). Overall 12 pts (50%) were CTC-positive at any time point, whereas 12 pts (50%) were consistently CTC-negative at both time points. A significant overall reduction in CTCs counts was observed from baseline to first evaluation (Sum of Ranks: 65 vs 13, p = 0.038, Wilcoxon t test). No associations were found between CTC detection or their kinetics, with clinicopathological features or treatment response. However, Kaplan Meier analysis revealed significantly reduced overall survival (OS) rates among pts with ≥ 5 CTCs at baseline (median OS: 18.0 versus 48.4 months; p = 0.010), at first evaluation (median OS: 18.0 versus 47.2 months; p = 0.000), or at any time point (median OS: 18.0 versus 47.2 months; p = 0.013). Conclusions: CTCs numbers significantly decreased during treatment in HR+ MBC pts receiving CDK4/6 inhibitors. The presence of ≥5 CTCs at any time point was strongly associated with significantly reduced overall survival, supporting their role as a negative prognostic biomarker in HR+ MBC.

BACKGROUND:Approximately 20-28% of NSCLC patients receiving ICIs also receive systemic antibiotics. Whether this reflects a true microbiome-mediated pharmacological interaction or confounding by indication remains unresolved. METHODS:We searched PubMed, Scopus, and CENTRAL (2013-2025) for NSCLC studies reporting OS, PFS, or ORR by antibiotic exposure. Random-effects meta-analysis was performed, with mostly non-pre-specified subgroup analyses by study design, ICI class, antibiotic timing, and line-of-therapy, plus leave-one-out, exposure-window, and adjustment-status sensitivity analyses and meta-regression of heterogeneity sources. Risk of bias was assessed with ROBINS-I and ROB 2, and certainty of evidence was rated with GRADE. RESULTS:Forty-one studies including 54,250 patients were analyzed, 27.8% of whom were antibiotic-exposed. Antibiotic exposure was associated with worse OS (HR 1.47, 95% CI 1.30-1.66; p < 0.00001; I2 = 79%) and PFS (HR 1.32, 95% CI 1.18-1.47; p < 0.00001; I2 = 65%), but not ORR (OR 0.95, 95% CI 0.67-1.35; p = 0.77; I2 = 88%). RCT post-hoc estimates were non-significant (OS HR 1.20, p = 0.07; PFS HR 1.09, p = 0.41; I2 = 0%), whereas observational estimates were larger and driven by less-adjusted cohorts. Subgroup, sensitivity, and meta-regression analyses indicated that heterogeneity was explained mainly by study design and line of therapy. In chemo-free ICI analyses, only mixed ICI regimens remained significant for OS and PFS. GRADE certainty was very low for observational outcomes and low for RCT-based estimates. CONCLUSIONS:The strongest available evidence does not support a clear antibiotic-related reduction in ICI efficacy in NSCLC. The observational signal is more compatible with confounding and exposure heterogeneity than with a uniform pharmacological interaction, although a context-specific biological effect cannot be excluded.

INTRODUCTION:HER2 mutations define a distinct, therapeutically actionable subset of NSCLC. Trastuzumab deruxtecan (T-DXd) has demonstrated strong activity in clinical trials, but real-world data are limited. METHODS:We conducted a retrospective, multinational cohort study of patients with advanced HER2-mutant NSCLC treated with T-DXd outside clinical trials between August 2021 and January 2025 across 68 centers in Europe and Israel. The primary end point was objective response rate (ORR). Secondary end points included progression-free survival (PFS), overall survival (OS), intracranial efficacy, and safety. RESULTS:Among 168 patients (median age 62 y; 59% female; 56% never-smokers), the ORR was 54.8% (95% confidence interval [CI]: 46.9-62.4) and disease control rate was 88.7% (95% CI: 82.9-93.1). Median PFS was 7.2 months (95% CI: 6.2-9.7) and median OS was 18.3 months (95% CI: 13.3-24.8). Treatment-naive patients (n = 18) achieved an ORR of 72.2% (95% CI: 46.5-90.3) and a median OS of 22.1 months (95% CI: 10.0-not evaluable). Patients with measurable brain metastases (n = 27) had an intracranial ORR of 74.1% (95% CI: 53.7-88.9), including 25.9% complete responses. Grade 3 or higher treatment-related adverse events occurred in 32% of patients. Interstitial lung disease/pneumonitis occurred in 14% (four fatal cases) of patients, without consistent predictors. CONCLUSIONS:In the largest real-world cohort reported to date, T-DXd demonstrated robust systemic and intracranial activity in HER2-mutant NSCLC, including treatment-naive patients and those with active brain metastases who were largely excluded from prior studies. Toxicity was consistent with previous reports, with interstitial lung disease remaining the main safety concern. These findings support integration of HER2-targeted therapies into evolving treatment algorithms.

OBJECTIVE:This study aimed to assess the cost-effectiveness of lung cancer screening (LCS) employing volume-based low-dose computed tomography (LDCT) in contrast to the absence of screening, targeting an asymptomatic high-risk population in Greece, leveraging the outcomes derived from the NELSON study, the largest European randomized control trial dedicated to LCS. METHODS:A validated model incorporating a decision tree and an integrated state-transition Markov model was used to simulate the identification, diagnosis, and treatments for a population at high risk of developing lung cancer, from a healthcare payer perspective. Screen-detected lung cancers, costs, life years (LYs), quality-adjusted life years (QALYs), and the incremental cost-effectiveness ratio (ICER) were predicted. Sensitivity and scenario analyses were conducted to assess the robustness and reliability of the model's outcomes under varying parameters and hypothetical situations. RESULTS:Annual LCS with volume-based LDCT detected 17,104 more lung cancer patients at early-stage among 207,885 screening population, leading to 8,761 premature lung cancer deaths averted. In addition, in contrast to no screening, LCS yielded 86,207 LYs gained and 50,207 incremental QALYs at an additional cost of €278,971,940, resulting in an ICER of €3,236 per LY and €5,505 per QALY, over a lifetime horizon. These estimates were robust in sensitivity analyses. CONCLUSIONS:LCS with volume-based LDCT, targeting an asymptomatic high-risk population, is highly cost-effective in Greece. Implementing LCS ensures efficient allocation of public healthcare resources while delivering substantial clinical benefits to lung cancer patients.

Background: Trophoblast cell surface antigen 2 (TROP2) promotes breast cancer (BC) development, invasion and metastasis, with promising role as a biomarker and therapeutic target in triple negative BC. Due to the dynamic tumor evolution during disease progression, a significant discordance in tumor cell profiles is frequently observed among primary tissues and distant metastases. In this regard, analyses of circulating tumor cells (CTCs) in the peripheral blood (PB) can inform on the expression of biomarkers in real-time. In the current study we assessed in parallel the expression of TROP2 on CTCs and matched primary tumors and metastatic sites from patients with triple negative BC. Methodology: PB was collected from 54 patients and CTCs were enriched by ficoll-density gradient centrifugation. Cytospins were immunofluorescently stained using antibodies for cytokeratins (Clones: AE1/AE3 & C11), CD45 and TROP-2 (Enzo Life Sciences); TROP2 expression on CTCs was defined as high, low or negative, by using the high TROP2-expressing MDA.MB.231 triple negative BC cell line as internal control. Matched primary (n=51) and metastatic (n=7) tumor tissue samples were evaluated for TROP-2 expression by immunocytochemistry (IHC); H-score was calculated as follows: (3 × % cells with strong intensity staining) + (2 × % cells with moderate intensity staining) + (1 × % cells with mild intensity staining), ranging from 0 to 300, and the following expression categories were defined: H-score 0 to <100: TROP2 low; H-score 100-200: TROP2 medium; H-score >200-300: TROP2 high. Results: CTCs (CK+/CD45- cells) were identified in 12 out of 54 patients evaluated (total CTC counts: n=80; mean CTC counts per patient: n=6.7). TROP2-expressing CTCs were detected in 75% of CTC-positive patients and represented the 95% of total CTCs. Specifically, high and low TROP2-expressing CTCs were identified in 66.7% and 41.7% of patients, representing the 81.3% and 13.7% of total CTCs, respectively. Differential TROP2 expression levels (high, medium and low) were also observed in both primary and metastatic tumors, showing a great intra-tumoral heterogeneity. High TROP2 expression was identified in 58.8% and 57.1% of primary and metastatic tissues, respectively. When matched primary and metastatic tissues were analyzed, a decrease in TROP2 expression was observed [median H-Score: 172.5 (range: 11-300) versus 87.5 (range: 5-150) in primary and metastatic tissue, respectively, p=0.068)]. CTC detection in the PB was not associated with TROP2 expression levels in primary or metastatic tissue (CTCs were identified in 13.3% and 50% of patients with high TROP2-expressing primary and metastatic tumors, respectively). Finally, there was no concordance in TROP2 expression pattern among CTCs and the respective tumor (high, low and negative TROP2-expressing CTCs were identified in 50%, 25% and 25% of patients with high TROP2-expressing primary tumors, respectively; high and low TROP2 -expressing CTCs were evident in all patients with high TROP2-expressing metastatic tumors). Conclusions: Herein we demonstrate for the first time a significant discrepancy in TROP2 expression among CTCs, primary and metastatic tumor tissue samples in triple negative BC. A lower TROP2 expression was observed in metastatic as compared to primary tissue, while no concordance was demonstrated among CTCs and the respective tumors. The results suggest the dynamic change in TROP2 expression status among different disease sites, thus highlighting the value of using liquid biopsy as a tool for real-time biomarker assessment in triple negative BC. Citation Format: Dimitrios Mavroudis, Eleni Lagoudaki, Sofia Gounaki, Sofia Hatziavraam, Charalampos Fotsitzoudis, Kleita Michaelidou, Sofia Agelaki, Maria A Papadaki. Comparative analysis of TROP2 expression in tumor tissues and circulating tumor cells (CTCs) in the peripheral blood of patients with triple negative breast cancer [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr P4-05-27.

Background Unless greater awareness inspires action, lung cancer will become the next global public health emergency. Despite improvements in screening, diagnosis, and treatments, more must be done because global incidence continues to increase. Concerted action must address the burden both between and within countries. Inequalities result from the uneven distribution of resources while inequities result from differences in prevention, screening, diagnosis, and treatment. Method Participants in the 9th International Lung Cancer Network conference (Athens, June 2023) received an advanced draft of the declaration with a confidentiality notice by email upon completion of online registration. They were invited to provide comments and suggestions up to seven days before the meeting. All contributions were reviewed by the authors and incorporated into the final version circulated the day before the meeting began. Although not a formal Delphi-style process, this approach prioritized inclusivity, transparency, and broad endorsement. Outreach extended beyond onsite attendees, ensuring diverse perspectives and representation. This process complements methodological consensus approaches by uniting stakeholders around shared policy goals. Results At the 9th International Lung Cancer Network conference, participants were invited to sign the final declaration, and 785 did so. Since then, 20 organizations worldwide have formally endorsed the Consensus Statement, and support continues to grow. The inclusive design has fostered broad and sustained backing, underscoring the value of the Statement as a unifying policy declaration advancing shared principles. Conclusion The Consensus Statement, Bridging the Gap in the Diagnosis and Management of Lung Cancer, reinforces five unifying principles: improve and expand prevention; focus on early detection; ensure equitable and sustainable access to treatment; build partnerships and promote investment; and, combat stigma, recognizing it as a social determinant of public health.

Breast cancer is classified, based on human epidermal growth factor receptor 2 (HER2) status, as HER2-positive or HER2-negative, guiding eligibility for HER2-targeted therapies. A HER2-low tumor is a type of breast cancer, which is defined by immunohistochemistry scores of 1+ or 2+ with negative in situ hybridization. Technological advances in HER2 testing, along with improved diagnostic precision, have increased the recognition of the HER2-low subgroup. HER2-low tumors may represent a distinct biological and clinical entity. Molecular profiling further demonstrates the unique transcriptomic and mutational signatures of HER2-low tumors, particularly within hormone receptor-positive contexts. In addition, clinical trials such as DESTINY-Breast04, demonstrate notable efficacies for novel antibody-drug conjugates, such as trastuzumab-deruxtecan, in patients with HER2-low tumors. This challenges the conventional HER2 stratification and suggests guideline revisions in order to acknowledge the HER2-low status. The present review synthesized knowledge regarding HER2 biology, pathology and molecular features, and highlighted the clinical relevance and therapeutic implications of redefining HER2 status to include the HER2-low subgroup. Recognizing HER2-low as a discrete classification may refine patient stratification, optimize therapeutic selection and improve clinical outcomes in breast cancer management.

Novel therapies have significantly improved survival for non-metastatic non-small cell lung cancer (NSCLC), however recurrence remains a challenge. Current treatment and surveillance strategies rely on imaging and clinical assessments with limited sensitivity in early detection of disease progression. Liquid biopsy-mediated detection of minimal residual disease (MRD) allows monitoring of tumor activity at the molecular level before clinical and radiologic progression. Here, we review the current evidence for MRD in the adaptive management and surveillance of non-metastatic NSCLC, focusing on the missing links that prevent its widespread clinical adoption.

e15026 Background: CDK4/6 inhibitors represent an important treatment option for early and metastatic breast cancer (BC). The detection of circulating tumor cells (CTCs) is a strong adverse prognostic factor in BC, however limited data exist on their prognostic value in patients receiving treatment with CDK4/6 inhibitors. We herein aimed to assess the prognostic value of CTCs, isolated with different assays, in patients treated with CDK4/6 inhibitors. Methods: Peripheral blood (PB) was obtained from 67 patients with BC (early stage: n = 9; metastatic stage: n = 58), prior to the start of CDK4/6 inhibitors plus endocrine treatment. CTCs were in parallel enriched using Ficoll density gradient centrifugation and the automated size-based Parsortix system (ANGLE plc). CTCs were identified via immunofluorescence staining with antibodies against cytokeratins (CKs)/CD45/dapi, followed by assessment using fluorescence microscopy. Results: CTCs (CK+/CD45- cells) were detected in 14.3% and 38.5% of patients with early and metastatic BC, respectively (mean CTC number per patient: n = 1.17 vs 3.15; p = 0.316). The enrichment of CTCs using different approaches, Ficoll and Parsortix, did not affect the overall CTC detection rate, which was 11.4% and 4.5% using the cut-off values of ≥2 CTCs and ≥5 CTCs, respectively. However, the positivity concordance rate between the two assays was only 6.8%. CTC detection was not associated with clinicopathological features or response to treatment. However, in the metastatic setting, a reduced overall survival (OS) was demonstrated among patients with detectable ≥2 CTCs and ≥5 CTCs using the Parsortix system (median OS: 15.5 versus 48.4 months; p = 0.000, and median OS: 11.2 versus 47.2 months; p = 0.000, respectively; Kaplan Meier analysis). Conclusions: CTCs are more frequently detected in patients with metastatic as compared to early stage BC. The Ficoll and Parsortix approaches provide similar CTC positivity rates, however they allow CTC detection mostly in different patients. The results also support an adverse prognostic value of CTCs identified by the Parsortix system for patients with metastatic BC treated with CDK4/6 inhibitors.

Background/Objectives: Alterations in DNA damage repair mechanisms can impair the therapeutic effectiveness of cisplatin. MicroRNAs (miRNAs), key regulators of DNA damage repair processes, have been proposed as promising biomarkers for predicting the response to platinum-based chemotherapy (CT) in non-small cell lung cancer (NSCLC). In this study, by using a bioinformatics approach, we identified six miRNAs, which were differentially expressed (DE) between NSCLC patients characterized as responders and non-responders to platinum-based CT. We further validated the differential expression of the selected miRNAs on tumor and matched normal tissues from patients with resected NSCLC. Methods: Two miRNA microarray expression datasets were retrieved from the Gene Expression Omnibus (GEO) repository, comprising a total of 69 NSCLC patients (N = 69) treated with CT and annotated data from their response to treatment. Differential expression analysis was performed using the Linear Models for Microarray Analysis (Limma) package in R to identify DE miRNAs between responders (N = 33) and non-responders (N = 36). Quantitative real-time PCR (qRT-PCR) was used to assess miRNA expression levels in clinical tissue samples (N = 20). Results: Analysis with the Limma package revealed 112 DE miRNAs between responders and non-responders. A random-effects meta-analysis further identified 24 miRNAs that were consistently up- or downregulated in at least two studies. Survival analysis using the Kaplan–Meier plotter (KM plotter) indicated that 22 of these miRNAs showed significant associations with prognosis in NSCLC. Functional and pathway enrichment analysis revealed that several of the identified miRNAs were linked to key pathways implicated in DNA damage repair, including the p53, Hippo, PI3K and TGF-β signaling pathways. We finally distinguished a six-miRNA signature consisting of miR-26a, miR-29c, miR-34a, miR-30e-5p, miR-30e-3p and miR-497, which were downregulated in non-responders and are involved in at least three DNA damage repair pathways. Comparative expression analysis on tumor and matched normal tissues from surgically treated NSCLC patients confirmed their differential expression in clinical samples. Conclusions: In summary, we identified a signature of six miRNAs that are suppressed in NSCLC and may serve as a predictor of cisplatin response in NSCLC.

e20654 Background: Non-small-cell lung cancer (NSCLC) is the most common histological subtype of lung cancer and the leading cause of cancer-related deaths worldwide. Targeted therapies and immunotherapies have significantly increased the treatment options for patients with advanced NSCLC and have also improved their progression-free survival (PFS). In this study molecular profiling in 4088 NSCLC patients, using NGS analysis, reveals actionable mutations in lung cancer and the eligibility of these patients in receiving targeted therapy. Methods: DNA and RNA extraction from embedded paraffin tissue samples was performed, using the Qiasymphony DSP DNA Mini Kit (Qiagen) and the RNeasy FFPE Kit (Qiagen) respectively. Μutation hotspot regions of 27 genes are amplified using a custom DNA panel (Thermo Fisher Scientific). Copy number variations, SNPs, and indels are analysed. Additionally, ALK, ROS1, RET, and NTRK1-3 fusions and expression and MET exon 14 skipping were tested using a custom Fusion Panel (Thermo Fisher Scientific). Sequencing was carried out using the Next Generation Sequencing platform Ion GeneStudio S5 Prime System (Thermo Fisher Scientific). Results : The NGS analysis detected actionable mutations in 92,6% of the tested samples, while 31,4% of these patients were eligible for on label therapy. The most common mutated gene was found to be KRAS (31,8%). The hotspot KRAS mutation G12C was detected in 10,3% of cases and it is associated with response to the FDA approved drugs Sotorasib and Adagrasib. The second most mutated gene was EGFR (12,1%). 11,7% of the patients harbored an EGFR mutation and were eligible for anti-EGFR therapies. Additionally, ALK fusion was detected in 2,2% of patients offering response to on label therapies like Brigatinib and Lorlatinib. Other findings that are associated with on label therapy include BRAF mutations (1,6%), MET exon 14 skipping (1,1%), ERBB2 mutations (0,8%), ROS1 fusions (0,8%) and RET fusions (0,5%). Finally, 61,6% of cases were also tested for PDL-1 expression, and 40,9% of them were predicted to have a positive response to immunotherapy. Notably, 20,8% of PDL-1 positive patients carried a mutation in STK11 gene, which is associated to resistance to immunotherapy. Conclusions: The findings described above underline the necessity of a multigene analysis for patients with NSCLC, in order to benefit from the available targeted therapies. PDL-1 testing increases this benefit as it is a positive predictive biomarker for immunotherapy, even for patients harboring no driver mutations.

The early diagnosis of lung cancer improves the probability of successful treatment. However, patients and physicians face several difficulties that can considerably delay the diagnostic process. A mixed-methods study that would follow the patient's journey throughout the diagnostic process could alleviate these difficulties. This study aimed to (a) track the patients' journey from the onset of symptoms until diagnosis and, (b) explore the patients' perspective of the journey until diagnosis, on the largest island of Greece. A convergent mixed-methods study was conducted with 94 patients with lung cancer. Patients completed a self-report questionnaire and were interviewed about their symptoms and journey through the healthcare system before their diagnosis. Our findings revealed several problems and delays in the diagnostic process. Both quantitative and qualitative data showed that patients did not recognize their symptoms and sought medical advice in time because they overlooked or attributed their symptoms to 'simpler'/'more common' causes. Furthermore, most patients were diagnosed 1-3 months after their first visit to a physician for their symptoms. Qualitative data analysis revealed three broad categories of problems that delayed diagnosis: (1) physician missteps, (2) administrative problems, and (3) the effect of the Covid-19 pandemic. This study found that major issues and delays prolong the diagnostic process for lung cancer. Therefore, optimization of diagnostic processes at each level of healthcare and interspecialty cooperation programs are needed. Furthermore, population-based interventions and patient education can help lung cancer patients be diagnosed early and improve their quality of life and disease outcomes.

MicroRNAs (miRNAs) are key regulators in immune surveillance and immune escape as well as modulators in the metastatic process of breast cancer cells. We evaluated the differential expression of plasma miR-10b, miR-19a, miR-20a, miR-126 and miR-155, which regulate immune response in breast cancer progression and we investigated their clinical relevance in the outcomes of breast cancer patients. Plasma samples were obtained from early (eBC; n = 140) and metastatic (mBC; n = 64) breast cancer patients before adjuvant or first-line chemotherapy, respectively. Plasma miRNA expression levels were assessed by qRT-PCR. We revealed a 4-miRNA panel consisted of miR-19a, miR-20a, miR-126, and miR-155 able to discriminate eBC from mBC patients with an AUC of 0.802 (p < 0.001). Survival analysis in eBC patients revealed that low miR-10b and miR-155 expression was associated with shorter disease free survival (disease free survival; p = 0.012 and p = 0.04, respectively) compared to high expression. Furthermore, miR-126 expression was associated with shorter overall survival (overall survival; p = 0.045). In multivariate analysis the number of infiltrated axillary lymph nodes and low miR-10b expression independently predicted for shorter DFS (HR: 2.538; p = 0.002 and HR: 1.943; p = 0.033, respectively) and axillary lymph nodes and low miR-126 for shorter OS (HR: 3.537; p = 0.001 and HR: 2.558; p = 0.018). In the subgroup of triple negative breast cancer (TNBC) patients, low miR-155 expression independently predicted for shorter DFS (HR: 5.056; p = 0.037). Accordingly in mBC, patients with low miR-10b expression had shorter progression free survival and OS compared to patients with high expression (p = 0.0017 and p = 0.042, respectively). In multivariate analysis, recurrent disease and low miR-10b expression independently predicted for shorter PFS (HR: 2.657; p = 0.001 and HR: 1.920; p = 0.017, respectively), whereas performance status two independently predicted for shorter OS (HR: 2.031; p = 0.03). In summary, deregulated expression of circulating miRNAs involved in tumor and immune cell interactions evaluated before adjuvant and 1st-line chemotherapy can distinguish disease status and emerge as independent predictors for outcomes of breast cancer patients.

The Fas/Fas ligand (FasL) system is a major apoptosis-regulating pathway with a key role in tumor immune surveillance and metastasis. The expression of Fas/FasL on mammary tumor tissues holds prognostic value for breast cancer (BC) patients. We herein assessed Fas/FasL expression on circulating tumor cells (CTCs) and matched peripheral blood mononuclear cells (PBMCs) from 98 patients with metastatic BC receiving first-line treatment. Fas+, FasL+, and Fas+/FasL+ CTCs were identified in 88.5%, 92.3%, and 84.6% of CTC-positive patients, respectively. In addition, Fas+/FasL+, Fas-/FasL+, and Fas-/FasL- PBMCs were identified in 70.3%, 24.2%, and 5.5% of patients, respectively. A reduced progression-free survival (PFS) was revealed among CTC-positive patients (median PFS: 9.5 versus 13.4 months; p = 0.004), and specifically among those harboring Fas+/FasL+ CTCs (median PFS: 9.5 vs. 13.4 months; p = 0.009). On the other hand, an increased overall survival (OS) was demonstrated among patients with Fas+/FasL+ PBMCs rather than those with Fas-/FasL+ and Fas-/FasL- PBMCs (median OS: 35.7 vs. 25.9 vs. 14.4 months, respectively; p = 0.008). These data provide for the first time evidence on Fas/FasL expression on CTCs and PBMCs with significant prognostic value for patients with metastatic BC, thus highlighting the role of the Fas/FasL system in the peripheral immune response and metastatic progression of BC.

BACKGROUND:Limited information exists on epidermal growth factor receptor (EGFR) molecular epidemiology in Greece. Next-generation sequencing (NGS) is the recommended method for EGFR genotyping in NSCLC. The Idylla Biocartis platform is a fully automated system for actionable EGFR mutation detection. RESEARCH DESIGN AND METHODS:We describe the prevalence of EGFR mutations in NSCLC patients in two high-volume clinical centers in Greece and compare key methods used for their determination. Eight hundred and fifty-seven FFPE samples from NSCLC patients were tested for EGFR mutations at University of Crete (UoC; n = 324) and at Evangelismos Hospital, Athens (Evangelismos; n = 503). RESULTS:The prevalence of EGFR mutations was 11.1% in the whole cohort (11.5% in non-squamous). The detection rate was 11.0% by NGS, 9.8% by Sanger and 11.3% by Idylla for the whole cohort (12.0% in non-squamous). The agreement between Idylla and Sanger was 93.2%. A targetable EGFR mutation was detected in 10.0% using tissue NGS alone, and in 16.0% using concurrent Idylla ctEGFR testing. CONCLUSION:The frequency of EGFR mutations was as expected for a Caucasian population. The Idylla EGFR test performance is comparable to reference methods and with a shorter TAT. Adding a concurrent plasma Idylla test to tissue NGS testing increases the detection rate of EGFR mutations in NSCLC.