Introduction Bone metastases (BMs) in patients with metastatic renal cell carcinoma (mRCC) negatively affect survival, quality of life, and increase the risk of skeletal-related events (SREs). Evidence remains limited in the era of first-line immune-based combinations. Patients and methods Meet-URO 33 is an Italian multicenter observational retrospective–prospective study enrolling mRCC patients receiving first-line therapy. The primary endpoint was overall survival (OS). Secondary endpoints included progression-free survival (PFS), clinical characterization, incidence of SREs, and impact of bone-targeting agents (BTAs). Survival was analyzed using the Kaplan–Meier method, log-rank test and Cox proportional hazards model. Results A total of 1696 patients enrolled between 2021 and 2025 were included; 526 (31%) had BMs at metastatic diagnosis. Patients with BMs more frequently had poorer performance status and unfavorable IMDC risk. The presence of BMs was associated with significantly worse OS (median 26.9 vs 102.1 months; HR 0.53, p<0.001) and worse PFS (median 14.2 vs 19.4 months; HR 0.71, p<0.001), with OS and PFS varying according to first-line regimen. Worse OS persisted across IMDC risk classes and treatment types; PFS differences were not significant in favorable/intermediate IMDC risk groups and with TKI monotherapy. Neither anatomical site nor number of BMs significantly affected OS. SRE incidence in patients with BMs was 26.8%, more frequent with spinal, rib, or other-site involvement, and more common in BTA-treated patients (22.2% vs 13.7%, p = 0.03), likely reflecting selection bias. Conclusions BMs are confirmed a negative prognostic factor in mRCC involving persistent unmet clinical needs.

ABSTRACT Aim This case series aimed to explore the occurrence of synchronous hepatocellular carcinoma (HCC) and gastrointestinal adenocarcinoma in cirrhotic patients and to propose a potential common pathogenic mechanism. Cases We reviewed the available literature and retrospectively analyzed seven cases of cirrhotic patients with synchronous HCC and gastrointestinal adenocarcinoma (colon or gastric) identified in our center between March 2020 and June 2023. All patients underwent upper gastrointestinal endoscopy, abdominal ultrasound, computed tomography (CT) scan, and histological confirmation through biopsy or surgery. The mean age of the patients was 77.3 years (range 76–83), with five males and two females. Five patients had liver cirrhosis, and two had chronic hepatitis (one with HCV, one with MASLD). HCC was confirmed in all patients, with elevated alpha‐fetoprotein levels (mean: 737.6 ng/mL). Colon adenocarcinoma was found in five patients, and gastric adenocarcinoma in one patient. Genetic and microsatellite instability analyses were performed in selected cases, revealing high microsatellite instability in one patient. We suggest that the Wnt/APC/β‐catenin pathway might play a key role in the pathogenesis of both HCC and gastrointestinal malignancies. Conclusions Synchronous HCC and gastrointestinal adenocarcinoma may be increasingly identified due to prolonged survival in cirrhotic patients. Alterations in the Wnt/APC/β‐catenin pathway could represent a shared pathogenic mechanism. Regular surveillance through ultrasound and endoscopy is essential for early diagnosis in this high‐risk population. Future research is needed to confirm these findings and explore targeted treatments.

e16539 Background: Cabozantinib is approved as first-line therapy for patients with intermediate-poor risk metastatic renal cell carcinoma (mRCC); however translating results from pivotal trials to an unselected real-world population remains a concern. Methods: CabFRONT is a retrospective observational multicentre Italian Network for Research in Urologic-Oncology (Meet-URO) study of a real-life population of treatment-naïve patients with intermediate-poor mRCC treated with cabozantinib as per clinical practice between September 2019 and September 2022. Results: 209 patients were enrolled. Median age was 65 years (36 - 85). 158 patients (76%) started with a dose of 60 mg, while 51 (24%) started with a dose of 40 mg. 140 patients (67%) were classified as intermediate and 69 (33%) as poor risk according to IMDC risk model. Clear-cell represented the most common histotype (172/209), followed by papillary (19/209), chromophobe (6/209) and NOS (4/209). Sarcomatoid features were present in 29 patients (14%). Lungs were the most frequent metastatic site, [137 (65%) patients], followed by lymph nodes [115 (55%)], and bones [72 (35%)]. 33 (16%) patients presented brain metastases. At a follow-up of 23 months (mo), the median progression-free survival (mPFS) was 9.6 mo (95% CI, 7.63-10.9 mo) and the median overall-survival was 21 mo (95% CI, 14-30 mo). As best overall response, 7 patients achieved a complete response, 73 a partial response and 77 stable disease for an objective response rate (ORR) of 38% and a disease control rate (DCR) of 75%. ORR for intermediate and poor population was respectively 29% and 10%, DCR was 56% for intermediate risk and 20% for poor risk population. mPFS for intermediate and poor population was respectively 12.73 and 5.27 mo (p < 0.0001). mPFS for intermediate with 1 risk factor was 17.2 and for intermediate with 2 risk factors was 9.7 mo (p 0.0033). Subsequent antineoplastic treatments were received by 41% (86/209) of patients. Subsequent nivolumab was received by 87% of patients. 191 patients (91%) reported at least 1 grade (G) 1 to 2 adverse event (AE). The most common G1 to G2 AEs were fatigue (113 [60%]), mucositis (100 [52%]), hand-foot syndrome (HFS) (91 [47%]), diarrhoea (79 [41%]), hypertension (67 [35%]), and hypothyroidism (60 [31%]). 78 G3 AEs ere reported: the most common were fatigue (19 [24%]) and diarrhoea (18 [23%]). Permanent discontinuations from the study owing to AEs was observed in 7 patients: three for G3 diarrhea, one of which manifested also G3 renal injury, two for G3 asthenia, one for G3 HFS and one for G3 mucositis. At least 131 patients (63%) required a dose reduction and 86 (41%) required a transitory interruption to manage toxic effects. Conclusions: Our data confirmed that front-line Cabozantinib is effective and safe in an unselected real-life population of patients with intermediate-poor mRCC, including non-clear cell histologies.

Background Evidence on homologous recombination repair (HRR) mutation prevalence in prostate cancer (PC) patients and the diagnostic testing path to guide treatment remains limited outside of clinical trials. The objective of this study was to investigate the DNA source, type of tumor tissue, timing for testing in the patient’s disease course, and rate of conclusive results in a real-world population. Patients and Methods This was an observational, cohort study, involving 20 Italian cancer centers. The study population included consecutive PC patients undergoing germline, tumor, and/or plasma circulating tumor DNA (ctDNA) to profile HRR genes between January 1, 2020 and January 31, 2025. Results Among 1400 PC patients included, 248 (17.7%) showed (likely)pathogenic variants (PVs) in the HRR genes. Most HRR testing was conducted during the metastatic castration-resistant PC (mCRPC)(779, 62.8%). The rate of conclusive results was 89.7% and varied widely according to the type of tumor tissue. The prevalence of HRR alterations was 18.1% in the mCRPC and 13.8% in the hormone-sensitive PC (P = .06). The concordance between tumor testing and ctDNA was 83.9%. Interestingly, 4.6% reported ctDNA testing positive but tumor testing negative, leading to important therapeutic implications. The prevalence of positive ctDNA testing was 47.4% vs 10.5%, for testing within 1 month or over 3 months, respectively, from the initiation of a new line of therapy. Conclusion This large real-world study, through the workflow adopted by clinicians for HRR genomic testing, provides novel insights into the variables influencing the success rate of genomic testing.

The aim of this study was to present our preliminary experience with transarterial radioembolization (TARE) using Yttrium-90 (90Y), compare the cancer-specific survival (CSS) of patients with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) liver metastases undergoing TARE, and investigate the influence of additional treatments on CSS. Our database was interrogated to retrieve patients who had undergone TARE using Yttrium-90 (90Y) glass or resin microspheres. Kaplan–Meier curves and the log-rank test were employed to conduct survival analysis for the different groups (p < 0.05). Thirty-nine patients were retrieved (sex: 27 M, 12 F; mean age: 63.59 ± 15.66 years): twenty-three with hepatocellular carcinoma (HCC) and sixteen with CRC liver metastasis. Globally, the patients with HCC demonstrated a significantly longer CSS than those with CRC liver metastasis (22.64 ± 2.7 vs. 7.21 ± 1.65 months; p = 0.014). Among the patients with CRC liver metastasis, those receiving TARE and additional concomitant treatments (n = 10) demonstrated a longer CSS than the CRC patients receiving only TARE (9.97 ± 2.21 vs. 2.59 ± 0.24 months; p = 0.06). In the HCC group, there was a trend of a longer CSS in patients (n = 8) receiving TARE and additional treatments (27.89 ± 3.1 vs. 17.69 ± 3.14 months; p = 0.15). Patients with HCC seem to achieve a longer survival after TARE compared to patients with CRC liver metastases. In patients with CRC liver metastases, the combination of TARE and additional concomitant treatments may improve survival.

We present our preliminary experience with transarterial radioembolization (TARE) with Yt-trium-90, and investigate the influence of additional treatments, injected activity, mean dose to the tumor on overall survival (OS). Our database was interrogated to retrieve patients who had undergone TARE with Yttrium-90 (90Y) glass or resin microspheres. The following information were searched: 1) type of pathology; 2) sex; 3) age; 4) administered activity; 5) mean dose to the tumor; 6) additional treatments; 7) OS. The OS of the different groups of patients were compared (p<0.05). A bivariate correlation (p<0.05) was used to investigate the association between injected activity and OS, and between mean dose to the tumor on OS. Thirty-nine patients were retrieved (Sex: 27 M, 12 F; mean age: 61.26 ± 14.95 years): 23 with hepatocellular carcinoma (HCC) and 16 with colorectal cancer (CRC) liver metastasis. Globally, patients with HCC demonstrated a sig-nificantly longer OS than those with CRC liver metastasis (22.66±19.11 vs. 10.41±8.75 months; p=0.022). Among patients with CRC liver metastasis, those receiving TARE and additional treatment demonstrated a longer OS than patients receiving only TARE (23.50±19.76 months vs. 8.54±5.31; p=0.018), there was a direct correlation between injected activity and OS (R=0.55, p=0.034) and between mean activity reaching the tumor and OS (R=0.812; p=0.008). Patients with HCC receiving TARE achieved a longer OS than those with CRC liver metastasis. Additional treatments, increasing injected activity and higher mean dose to the tumor seem beneficial for outcome, especially in patients with CRC liver metastases.

Abstract Background in the recent years, survival of cancer patients increased enormously through the use of new anticancer drugs such as molecular target drugs (tyrosine kinase inhibitors- TKIs, immunotherapy). Immunotherapy includes anti-BRAF and anti-MEK drugs (although evidences are scarce, these drugs seem to be capable of causing cardiovascular toxicity too); TKIs includes inhibitors of VEGF, VEGFR and other kinases. Purpose to assess cardiovascular toxic effects of TKIs and immunotherapy, to identify early signs of cardiac and vascular toxicity using speckle tracking echocardiography and arterial stiffness measurement. Methods a prospective study was carried out evaluating 45 patients treated with immunotherapy or TKIs. Population was divided into 2 groups: Group A (17 patients with melanoma treated with anti- BRAF and anti-MEK) and Group B (28 patients with solid cancer treated with TKIs-anti VEGF). Cardiological evaluation including electrocardiogram, conventional echocardiogram with tissue Doppler imaging (TDI) and left ventricular global longitudinal strain (GLS) measurement and carotid ultrasound scan was carried out before starting therapy and at a follow-up time of 6 and 12 months. Cardiovascular events such as heart failure, arterial hypertension, arrhythmias, Qtc interval prolongation, stroke, arterial and venous thrombosis were assessed during follow-up. Results mean follow-up was 7 months for anti-BRAF and 13 months for TKI. Neither cardiovascular adverse events nor significant reductions in LVEF or other echocardiographic parameters during follow-up were observed in Group A: we did not observe significant changes in GLS (-18,4 [RI -19,9 a -15,8] vs. -18,3 [RI -18,9 a -16,1] or in vascular parameters (PWV 7,3 ± 1,4 vs. 6,7 ± 1,5, β mean wave 8,5 [RI 6,3-12] vs. 7,4 [RI 4,9-9,4]; α mean wave 6,8 [RI 3,1-6,2] vs. 9,6 [RI 2,4-4,6]). As regards Group B, new onset of arterial hypertension in 35% of population but no significant changes in LVEF or other echocardiographic parameters including GLS ( -18,6 ± 2,3 vs. -18,3 ± 3,3) were observed. Significant changes were furthermore found in vascular parameters with increased arterial stiffness during follow-up (PWV 7,82 ± 1,23 vs. 10,02 ± 2,73; p = 0,04, β mean wave 5,33 ± 1,98 vs. 8,43 ± 3,03; p = 0,02, α mean wave 10,79 ± 4,03 vs.16,82 ± 6,38; p = 0,03). Conclusions according to the literature data, immunotherapy seems to have a safe cardiovascular profile. Anti-VEGF drugs seem to predominantly cause vascular damage, early identifiable through arterial stiffness measurement.

BACKGROUND:Target therapy can cause various cardiovascular complications. The aim of this study was to evaluate the burden of cardiovascular complications related to treatment with anti-BCR-ABL tyrosine kinase inhibitors (TKIs) and to determine if there are differences between the latest- and first-generation TKIs.METHODS:A retrospective observational study was carried out on 55 patients (39 men, 16 women; mean age ± SD: 58 ± 11 years) treated with TKIs targeting Bcr-Abl for a median period of 3.5 years. Patients were divided in two groups according to the type of treatment. Group A included patients treated with latest-generation TKI (nilotinib, dasatinib, and ponatinib), while group B included patients treated with first-generation TKI (imatinib). Cardiological evaluation included electrocardiogram, echocardiogram with global longitudinal strain of left ventricle (GLS), and carotid ultrasound scan with arterial stiffness measurement (pulse wave velocity, PWV). Adverse cardiovascular events were recorded in both groups.RESULTS:Statistical analysis showed that cardiovascular adverse events (myocardial ischemia, peripheral artery disease, deep vein thrombosis, and pleural effusion) were significantly more frequent in group A than group B (p value = 0.044). Moreover, there was a significant reduction in GLS and PWV in group A when compared to group B (respectively, p = 0.03 and p = 0.004).CONCLUSIONS:Our study confirms that imatinib is a relatively safe drug, while it reveals that the latest-generation TKIs may cause a burden of cardiovascular complications. GLS and PWV allow detection of early signs of cardiac and vascular toxicity in oncohematologic patients treated with TKI, and their use is advisable.

The identification of tumor "oncogenic drivers" and the subsequent development of targeted therapy represented a milestone in the treatment of lung cancer over the last years. Tumor genotyping has been incorporated into therapeutic decision making of advanced non-small cell lung cancer (NSCLC) since has become clear that individuals with actionable molecular alterations receiving a matched targeted agent certainly live longer and better. The recent understanding of biological mechanisms underlying cancer immune evasion has allowed the development of a new class of immunomodulatory agents which are able to reactivate host immune-response, offering the potential for long-term disease control and survival in a significant subgroup of lung cancer patients. The complementary therapeutic effects of these two different approaches suggested intriguing potential for therapeutic synergy with combination strategies. Indeed, immunotherapy could consolidate the dramatic but transient tumor responses achieved with targeted therapy into long-term survival benefit, due to the induction of specific anti-tumor memory. However, the great emphasis and expectations linked to immune-targeted combinations have been mostly disappointed by the initial controversial results of early-phase trials, raising relevant concerns about the use of these combinations for lung cancer treatment. This review briefly summarizes the basis of immunogenicity and immune escape in oncogene addicted NSCLC, providing an updated overview of clinical trials, with the final aim of defining the current unmet needs of immuno-targeted combinations in clinical practice.

The Cardio-oncology field has grown considerably in the last two decades. The remarkable increase in the number of molecules used in oncology has brought with it a huge set of cardiovascular adverse events. For this reason, it is necessary to intervene on the early stages of drug development. This is what the Food and Drug Administration aims to do. This purpose can be achieved through a more careful analysis of the adverse event, development of guidelines, and identification of objective parameters that could guide the researcher in defining precisely the adverse event. It is also necessary to use additional methods not yet used in clinical trials that can allow an early detection of adverse events and to highlight subclinical damage. These measures will allow the researcher to intervene and treat them even before it can expose the patient to more serious complications or the drug can determine deaths in the post-marketing phase for unacknowledged cardiovascular adverse events.

Vascular endothelial growth factor (VEGF) plays an important role in maintaining the regular homeostasis of vascular walls. VEGF binds its receptor (VEGFR) promoting the regular survival and function of endothelial cells. Anti-VEGF and anti-VEGFR drugs inhibit the action of VEGF and VEGFR. These drugs can cause cardiovascular toxic effects such as arterial hypertension, thromboembolism, myocardial ischemia and heart failure. The monoclonal antibody bevacizumab and tyrosine kinase inhibitors (sorafenib, sunitinib, pazopanib, regorafenib, axitinib, cabozantinib, ponatinib) are the main inhibitors of VEGF, VEGFR and other tyrosine kinases. In this chapter we will illustrate the cardiovascular toxic effects of these drugs, their mechanism of action, strategy to early diagnose and treat these complications. We will also illustrate strategy to prevent cardiovascular toxicity. It is important to know cardiovascular toxic effect of these drugs widely used in oncological field, to avoid the development of severe future complications.

Many new drugs have appeared in last years in the oncological treatment scenario. Each drug carries an important set of adverse events, not less, cardiovascular adverse events. This aspect is even more important considering the increasing use of combination therapies with two drugs, or three drugs as in some ongoing clinical trials. Besides it represents a growing problem for Cardiologists, that face it in every day clinical practice and that will face it probably more and more in the coming years. This work reviews the mechanism of action of BRAF-inhibitors and MEK-inhibitors used together, the pathophysiological mechanisms that lead to cardiovascular toxicity. Particularly, it focuses on hypertension and ejection fraction reduction development. Then, it follows the examination of published data for each combination therapy. A Literature research was carried out using Pubmed selecting review articles, original studies and clinical trials, but mainly focusing on phase 3 studies. This work aims to summarize the knowledge about BRAF-inhibitor and MEK-inhibitor treatment and its cardiovascular toxicity to make it usable and give the basic tools to Cardiologists and Oncologists for a better management of cancer patient undergoing this treatment. Besides a deeper knowledge of the cardiovascular adverse events linked to this treatment and the magnitude of their expression and frequency can lead to a targeted cardiological treatment.

The identification of oncogenic driver mutations in non-small-cell lung cancer (NSCLC) has led to the development of targeted drugs. Tyrosine kinase inhibitors (TKIs) directed against the epidermal growth factor receptor (EGFR) target lung tumours bearing EGFR-activating mutations. This new therapeutic strategy has greatly improved tumour response rates. However, drug resistance invariably occurs during TKI-based treatment. Epithelial-to-mesenchymal transition (EMT) is one of the resistance mechanisms identified in EGFR-mutated NSCLC treated with TKIs. In this review we gather together the most important findings on this phenomenon in relation to cancer stem cells and cancer epigenetics. We also outline the correlation between the effects of stromal factors from the microenvironment, the transcription factors activated, the epigenetic changes in chromatin, and the evolution of cellular behaviour. Notably, EMT has already been shown to be the link between benign lung diseases such as chronic obstructive pulmonary disease and lung carcinogenesis. The various mechanisms of acquired resistance to EGFR-TKIs are also briefly described to provide background information on EMT. Our extensive review of the scientific literature serves to highlight the cellular and molecular events that lead to the onset of EMT in NSCLC cells treated with EGFR-TKIs. Finally, we put forward a hypothesis to explain why, in some cases, EMT rather than other known mechanisms is involved in resistance to TKIs.

Angiogenesis is a key moment in tumor development and proliferation. Until recently oncologists did not know the mechanisms that were behind this phenomenon, but following the discoveries of Folkman and coworkers, they have gradually created and developed a series of drugs that act against angiogenesis by interacting with molecules belonging to the vascular endothelial growth factor (VEGFs) class and its receptors (VEGFRs) giving rise to anticancer effects. Tyrosine kinase inhibitors (TKIs) are a major class of these new anticancer agents, demonstrating high antitumor activity in a variety of “orphan” neoplasms (such as hepatocellular carcinoma, kidney cancer, sarcomas, etc.). The mechanism of action of these drugs also explains their toxicity profile with respect to the cardiovascular system. The aim of this chapter is to offer to the reader an updated overview of the new treatments available, with special attention to the assessment and management of cardiovascular side effects.

Background: Long non-coding RNAs (lncRNAs) are emerging as essential regulators of genetic and epigenetic networks. Their deregulation may underlie carcinogenesis suggesting their potential involvement in tumorigenic and metastatic processes, as well as their role as prognostic/predictive biomarkers for clinical use in patients with several solid tumors. Few studies evaluated lncRNAs expression in rare tumors such as Gastrointestinal Stromal Tumors (GISTs). Indeed, the upregulation of HOTAIR has been associated with tumor aggressiveness and metastasis, and poor survival of GIST patients. In order to gain more detailed insight on the molecular role of lncRNAs, we analyzed the expression levels of lncRNAs H19, MALAT1 and HOTAIR in tissue specimens of surgically resected GIST patients to evaluate the potential role of lncRNAs as prognostic biomarkers. Methods: The expression of the lncRNAs H19, MALAT1 and HOTAIR was evaluated in a total of 40 pairs of disease formalin-fixed paraffin-embedded tissue and adjacent normal tissue from 40 GIST patients with localized and locally advanced disease using quantitative real-time reverse transcriptase. Results: H19 was overexpressed in 50% GIST patients (p-value: 0.0496). MALAT1 was overexpressed in 45% GIST patients (p-value: 0.032). None of them had the related date with HOTAIR. Furthermore, the up-regulation of H19 has been found in 74% patients harboring cKIT mutations compared to 57% wild type patients (p-value: 0.042). Conversely the up-regulation of MALAT1 has been found in 76% patients harboring cKIT mutations compared to 100% wild type patients (p-value: 0.027). Finally, the up- regulation of H19 has been found in 100% patients with TTP < 3 months compared to 25% patients with TTP >3 months, while the up-regulation of MALAT1 has been found in 25% patients with TTP < 3 months compared to 75% patients with TTP >3 months. Conclusions: H19 and MALAT1 appear upregulated in GIST patients according to the KIT- mutation status. These data would suggest a potential opposite prognostic value of both H19 and MALAT1 lncRNAs in these patients. The results of HOTAIR expression levels were indeterminated in all analyzed tumor samples, probably because HOTAIR has been degraded during its isolation. Further analyses are needed to confirm these data. Legal entity responsible for the study: University of Palermo Funding: None Disclosure: All authors have declared no conflicts of interest.

BACKGROUND Recent studies evaluated the diagnostic accuracy of circulating tumor DNA (ctDNA) analysis in the detection of epidermal growth factor receptor (EGFR) mutations from plasma of NSCLC patients, overall showing a high concordance as compared to standard tissue genotyping. However it is less clear if the location of metastatic site may influence the ability to identify EGFR mutations. OBJECTIVE This pooled analysis aims to evaluate the association between the metastatic site location and the sensitivity of ctDNA analysis in detecting EGFR mutations in NSCLC patients. METHODS Data from all published studies, evaluating the sensitivity of plasma-based EGFRmutation testing, stratified by metastatic site location (extrathoracic (M1b) vs intrathoracic (M1a)) were collected by searching in PubMed, Cochrane Library, American Society of Clinical Oncology, and World Conference of Lung Cancer, meeting proceedings. Pooled Odds ratio (OR) and 95% confidence intervals (95% CIs) were calculated for the ctDNA analysis sensitivity, according to metastatic site location. RESULTS A total of ten studies, with 1425 patients, were eligible. Pooled analysis showed that the sensitivity of ctDNA-based EGFR-mutation testing is significantly higher in patients with M1b vs M1a disease (OR: 5.09; 95% CIs: 2.93 - 8.84). A significant association was observed for both EGFR-activating (OR: 4.30, 95% CI: 2.35-7.88) and resistant T790M mutations (OR: 11.89, 95% CI: 1.45-97.22), regardless of the use of digital-PCR (OR: 5.85, 95% CI: 3.56-9.60) or non-digital PCR technologies (OR: 2.96, 95% CI: 2.24-3.91). CONCLUSIONS These data suggest that the location of metastatic sites significantly influences the diagnostic accuracy of ctDNA analysis in detecting EGFR mutations in NSCLC patients.

Background: Oral squamous cell carcinoma (OSCC) is the most frequent malignant tumor of the oral cavity with low survival rate, accounting for more than 95% of all head and neck cancers. Generally, fewer than 50% of patients survives more than 5 years, because this tumor is often detected at a late stage. Since saliva has been shown to be a non-invasive, accessible, and highly efficient diagnostic medium, the analysis of salivary biomarkers may provide an efficient tool for oral cancer early detection. The main aim of our study was to analyze the salivary microRNA expression profile in OSCC patients, in order to investigate the molecular mechanisms and signaling pathways responsible for the development and progression of this tumor. Materials and methods: Total RNA and miRNAs were isolated using the miRNeasy Mini Kit, and their quality and quantity were assessed using the 2100 Bioanalyzer and spectrophotometer NanoDrop ND-1000. Using a TaqMan Low Density Array A human microRNA microarray analysis, the expression profile of 377 miRNAs was analyzed in saliva of ten OSCC patients and ten healthy individuals. In addition, the expression of specific salivary miRNAs was analyzed in other independent samples from fifteen OSCC patients at different stages of disease, using Real-time PCR analyses. Results: Microarray analysis showed that a subset of twelve miRNAs, such as let-7g, miR-27a, miR-30b, miR-133a, miR-135b, miR-148b, miR-183, miR-199a-3p, miR-328, miR-361-5p, miR-451 and miR-486-5p, involved in several cancer-related pathways, including TGF-b, PI3K/Akt, Wnt, MAPK signaling, was differentially expressed in saliva of OSCC patients. Among these deregulated salivary miRNAs, eight were found to be up-regulated in their expression and four down-regulated. Finally, expression analysis of hypothetical miRNA gene targets involved in the same cancer-related pathways confirmed the coherence of our results. Conclusions: Recent findings reported in literature regarding variations in expression of some miRNAs involved in proliferation, metastasis development, and therapy response in OSCC have confirmed the coherence of our results.

Background: Low Intensity Pulsed Ultrasound (LIPUS) application has been shown to have an encouraging effect in inducing a transient pore formation through cellular membranes. This permeability condition has been demonstrated to be useful in enhancing gene and drug delivery. Nowadays, in the management of NSCLC patients, the use of liquid biopsy has entered the clinical practice. One of the main limits in the analysis of circulating tumor DNA is the low concentration rate of nucleic acids in body fluids. Ultrasound stimulation (US) has been recently demonstrated to be effective for the release of specific circulating tumor biomarkers in many mouse models. We demonstrated the role of US in inducing the release of tumor DNA fragments (rtDNA) in NSCLC without inducing any apoptotic or necrotic event. Material and methods: EGFR wt and del19 NSCLC cells (A549, HCC827) were cultured in RPMI1640 with 10% FBS, 1% pen/strep at 37 °C and 5% CO2. The day before the sonication cells were seeded in a 24-well plates (20.000 cells/well HCC827; 10.000 cells/well A549). Each well was exposed at the following sonication protocol: frequencies (650 kHz, 1 MHz); acoustic pressure (250 kPa, 25 kPa); 25% duty cycle with three different exposure time points (1, 3, 7.5 min). After US treatment, the cells were incubated for 24h and then cell viability was performed by Cell Titer-Glo® Luminescent Cell Viability Assay. Each experiment was performed in triplicate. Results: NSCLC cells have been subjected to sonoporation at different exposure time points as well as ultrasonic acoustic pressures and frequencies. We evaluated the viability cells to exclude the possibility that rtDNA analysis could be affected by apoptotic or necrotic DNA fractions. In fact, after ultrasound exposure no significant reduction of cell viability, in terms of ATP content, has been shown. Moreover, the analysis of DNA fragments content, released in the medium, showed a different behaviour on the basis of EGFR mutational status. Indeed, in EGFR mutated cells the concentration of rtDNA was significantly higher than control cells after performing sonoporation at 250 kPa and 1 MHz. For the EGFR wild type cell line, no variation of ctDNA at different exposure time points and pression has been showed. Conclusions: the assessment of ctDNA is strongly influenced by its amount. Therefore, US application to enhance the release, is of great interest not only in NSCLC but also for all the "oncogene addicted" cancers.

3565 Background: Clinical trials investigated the potential role of both KRAS and BRAF mutations, as prognostic biomarkers, in colorectal cancer (CRC) patients who underwent surgical treatment of liver metastasis (CLM), showing conflicting results. This meta-analysis aims to review all the studies reporting survival outcomes (recurrence free survival (RFS), and/or overall survival (OS)) of patients undergoing resection of CLM, stratified according to KRAS and/or BRAF mutation status. Methods: Data from all published studies reporting survival outcomes (RFS and/or OS) of CRC patients who received resection of CLM, stratified by KRAS and/or BRAF mutation status were collected by searching in PubMed, Cochrane Library, American Society of Clinical Oncology and European Society of Medical Oncology meeting proceedings. Pooled hazard ratios (HRs) and 95% confidence intervals (95% CIs) were calculated for both the OS and/or RFS. Results: Seven eligible trials (1403 patients) were included. Pooled analysis showed that KRAS mutations predicted a significant worse both RFS (HR: 1.65; 95% CI: 1.23 – 2.21) and OS (HR: 1.86; 95% CI: 1.51 – 2.30) in patients who underwent surgical resection of CLM. BRAF mutations were also associated with a significant worse OS (HR: 3.90; 95% CI: 1.96 – 7.73) in this subgroup of patients. Conclusions: This meta-analysis suggests both KRAS and BRAF mutations as negative prognostic biomarkers associated with worse survival outcomes in patients undergoing hepatic resection of CLM. Such evidences support the introduction of new treatment decision models, taking into account the tumor molecular profile in order to individualize both systemic and loco-regional treatment strategies.