Search

PURPOSE Limited prospective data are available on sequential immunotherapy and BRAF/MEK inhibition for BRAFV600-mutant metastatic melanoma. METHODS SECOMBIT is a randomized, three-arm, noncomparative phase II trial (ClinicalTrials.gov identifier: NCT02631447 ). Patients with untreated, metastatic BRAFV600-mutant melanoma from 37 sites in nine countries were randomly assigned to arm A (encorafenib [450 mg orally once daily] plus binimetinib [45 mg orally twice daily] until progressive disease [PD] -> ipilimumab plus nivolumab [ipilimumab 3 mg/kg once every 3 weeks and nivolumab 1 mg/kg once every 3 weeks × four cycles -> nivolumab 3 mg/kg every 2 weeks]), arm B [ipilimumab plus nivolumab until PD -> encorafenib plus binimetinib], or arm C (encorafenib plus binimetinib for 8 weeks -> ipilimumab plus nivolumab until PD -> encorafenib plus binimetinib). The primary end point was overall survival (OS) at 2 years. Secondary end points included total progression-free survival, 3-year OS, best overall response rate, duration of response, and biomarkers in the intent-to-treat population. Safety was analyzed throughout sequential treatment in all participants who received at least one dose of study medication. RESULTS A total of 209 patients were randomly assigned (69 in arm A, 71 in arm B, and 69 in arm C). At a median follow-up of 32.2 (interquartile range, 27.9-41.6) months, median OS was not reached in any arm and more than 30 patients were alive in all arms. Assuming a null hypothesis of median OS of ≤ 15 months, the OS end point was met for all arms. The 2-year and 3-year OS rates were 65% (95% CI, 54 to 76) and 54% (95% CI, 41 to 67) in arm A, 73% (95% CI, 62 to 84) and 62% (95% CI, 48 to 76) in arm B, and 69% (95% CI, 59 to 80) and 60% (95% CI, 58 to 72) in arm C. No new safety signals emerged. CONCLUSION Sequential immunotherapy and targeted therapy provide clinically meaningful survival benefits for patients with BRAFV600-mutant melanoma.

Background: Brain metastases (BMs) is one of the most frequent metastatic sites for non-small-cell lung cancer (NSCLC). It is a matter of debate whether EGFR mutation in the primary tumor may be a marker for the disease course, prognosis, and diagnostic imaging of BMs, comparable to that described for primary brain tumors, such as glioblastoma (GB). This issue was investigated in the present research manuscript. Methods: We performed a retrospective study to identify the relevance of EGFR mutations and prognostic factors for diagnostic imaging, survival, and disease course within a cohort of patients affected by NSCLC-BMs. Imaging was carried out using MRI at various time intervals. The disease course was assessed using a neurological exam carried out at three-month intervals. The survival was expressed from surgical intervention. Results: The patient cohort consisted of 81 patients. The overall survival of the cohort was 15 ± 1.7 months. EGFR mutation and ALK expression did not differ significantly for age, gender, and gross morphology of the BM. Contrariwise, the EGFR mutation was significantly associated with MRI concerning the occurrence of greater tumor (22.38 ± 21.35 cm3 versus 7.68 ± 6.44 cm3, p = 0.046) and edema volume (72.44 ± 60.71 cm3 versus 31.92 cm3, p = 0.028). In turn, the occurrence of MRI abnormalities was related to neurological symptoms assessed using the Karnofsky performance status and mostly depended on tumor-related edema (p = 0.048). However, the highest significant correlation was observed between EGFR mutation and the occurrence of seizures as the clinical onset of the neoplasm (p = 0.004). Conclusions: The presence of EGFR mutations significantly correlates with greater edema and mostly a higher seizure incidence of BMs from NSCLC. In contrast, EGFR mutations do not affect the patient’s survival, the disease course, and focal neurological symptoms but seizures. This contrasts with the significance of EGFR in the course and prognosis of the primary tumor (NSCLC).

http://www.breakthroughcentre.org.uk/jones_et_al/ (Table S1, Figure S2, Figure S3, Table S4)

4-year survival data of clinical trial SECOMBIT

SummaryIt has been estimated that 15%-20% of human cancers are attributable to infections, mostly by carcinogenic viruses. The incidence varies worldwide, with a majority affecting developing countries. Here, we present a comparative analysis of virus-positive and virus-negative tumors in nine cancers linked to five viruses. We find that virus-positive tumors occur more frequently in males and show geographical disparities in incidence. Genomic analysis of 1,658 tumors reveals virus-positive tumors exhibit distinct mutation signatures and driver gene mutations and possess a lower somatic mutation burden compared to virus-negative tumors of the same cancer type. For example, compared to the respective virus-negative counterparts, virus-positive cases across different cancer histologies had less often mutations ofTP53and deletions of 9p21.3/CDKN2A-CDKN1A; Epstein-Barr virus-positive (EBV+) gastric cancer had more frequent mutations ofEIF4A1andARID1Aand less marked mismatch repair deficiency signatures; and EBV-positive cHL had fewer somatic genetic lesions of JAK-STAT, NF-κB, PI3K-AKT and HLA-I genes and a less pronounced activity of the aberrant somatic hypermutation signature. In cHL, we also identify germline homozygosity in HLA class I as a potential risk factor for the development of EBV-positive Hodgkin lymphoma. Finally, an analysis of clinical trials of PD-(L)1 inhibitors in four virus-associated cancers suggested an association of viral infection with higher response rate in patients receiving such treatments, which was particularly evident in gastric cancer and head and neck squamous cell carcinoma. These results illustrate the epidemiological, genetic, prognostic, and therapeutic trends across virus-associated malignancies.

Anterior cranial fossa (ACF) dural arteriovenous fistulas (ACF DAVFs) are an infrequent subtype of cranial DAVFs that are usually fed by the anterior ethmoidal artery (AEA) and ophthalmic artery (OA) branches. Due to the lack of dural venous sinus in the ACF, they directly drain into cortical veins, resulting in high bleeding tendency. For this reason, ACF DAVFs have to be treated regardless of whether they are symptomatic or not. A 74-year-old man, with hypertension history, came to our attention because of ear pain, dizziness, and impaired hearing for 2 months. No other pathological conditions have been found in his medical history. The patient underwent brain magnetic resonance imaging (MRI) scan and subsequently second level diagnostic investigations with digital subtraction angiography (DSA), showing a foramen caecum (FC) patency and a persistent, enlarged, arterialized FC vein (FCV) involved in an incidental ACF DAVF (Cognard IV), fed mainly by sphenopalatine arteries (SPAs) branches bilaterally. The patient underwent open surgery performed by small high frontal craniotomy with DAVFs complete closure and without any complications. To the best of our knowledge, this is the first case ever described of FCV persistence with involvement in an intracranial vascular malformation. It has been managed by surgical intervention that can be considered, despite the large skin incision, a minimally invasive approach with an excellent cosmetic result and minimal risk of complications.

Background: Elevated human epididymis secretory protein 4 (HE4) serum levels have been widely investigated in patients with ovarian cancer. However, high levels of HE4 can be also found in other tumors and in renal fibrosis. To date, the HE4 assay manufacturer features the reference value only for the female pre- and post-menopausal population. The aim of this study was to determine the upper reference limit (URL) of HE4 in a well-defined and large cohort of healthy male individuals and investigate potential factors influencing HE4 levels in healthy subjects. Methods: The study included 307 Italian healthy male individuals. HE4 was measured using a chemiluminescent assay (Abbott Laboratories, Wiesbaden, Germany). The URL was calculated using the non-parametric percentile method. Differences in HE4 concentrations according to age, estimated glomerular filtration rate (eGFR), free and bioavailable testosterone were also evaluated. Results: The 97.5th percentile URL of serum HE4 in our study population was 57 pmol/L (90% CI). After stratifying subjects according to age, we found that the URL of HE4 was higher in older (> 50 years) than in younger subjects (18-30 years old), and overlapping with the URL in males from 31 to 50 years old (P=4.769e-16, r=0.44). A strong negative correlation between HE4 and eGFR was observed (P=8.412e-12, r=- 0.38). Moreover, a statistically significant negative correlation was also found between HE4 and free and bioavailable testosterone. Conclusion: In this study was determined the URL of HE4 in a large cohort of healthy male subjects. Our findings indicate that the HE4 age-dependent differences in males need to be taken into account. The definition of the HE4 URL in males and the correlation observed with eGFR and testosterone should foster the clinical use of HE4 beyond gynecologic cancer.

Background: Brain metastases (BMs) are the most common intracranial tumors in adults and the brain is one of the most frequent metastatic sites for non-small-cell lung cancer (NSCLC). Some studies have hypothesized that EGFR expression in the primary tumor may result in clinically radiological and prognostic changes in related brain metastases, comparable to EGFR's correlation with prognosis in primary brain tumors. There are no current studies that clinically and radiologically demonstrate a difference between BMs from EGFR-mutated and wild-type NSCLC.Methods: We performed a retrospective study to identify prognostic factors for the survival of patients with NSCLC-BMs by exploring the role of driving mutations in NSCLC, focusing on EGFR mutated status, and comparing all morphological, radiological, and clinical features of NSCLC-BMs with their outcome parameters in a cohort of surgically treated patients. Results: The final cohort consisted of 81 patients. The overall survival of the cohort was 15±17 months. The tumor-related edema was associated with neurological symptoms at the clinical onset of the disease (p=.048). We found that EGFR and ALK mutation status did not reach significant associations with age, sex, and the morphology of the lesions. EGFR mutation in the primary tumor is positively associated with higher edema and tumor volume (respectively 22.38±21.35cm3 versus 7.68±8.44cm3 and 72.44±60.71cm3 versus 31.92cm3 p=.046 and p=.028). Moreover, EGFR mutation is associated with a clinical debut of seizures (p=.004).Conclusions: The role of EGFR and ALK mutations of NSCLC on prognostic characteristics of BMs is still to be fully clarified; however the results retrieved from the present study suggest that the presence of EGFR mutations significantly correlates with bigger edema volume and higher incidence of seizures. In recent studies, while EGFR mutation status did not affect the immune pathway scores of primary lung tumors, the overall immune pathway scores in related BMs indicate a peculiar immunogenic phenotype that could explain a large amount of edema volume. This study could be considered the first clinical and radiological demonstration of this immunological phenotype.

Supplemental Digital Content is available in the text., Gastrointestinal stromal tumor (GIST) is the most common mesenchymal malignancy of the gastrointestinal tract. We provide in the present article the molecular characterization of a series of primary GISTs in a cohort of Sardinian patients (Italy), with the aim to describe the patterns of KIT and PDGFRa mutations and the corresponding clinical features. Ninety-nine Sardinian patients with histologically-proven diagnosis of GIST were included in the study. Medical records and pathology reports were used to assess the demographic and clinical features of the patients and the disease at the time of the diagnosis. Formalin-fixed, paraffin-embedded tissue samples were retrieved for each case, and mutation analysis of the KIT and PDGFRa genes was performed. KIT and PDGFRa mutations were detected in 81.8% and 5% of the cases, respectively. The most common KIT mutation was W557_K558del in exon 11, while D842V in exon 18 was the most common PDGFRa genetic alteration; V561D was the only PDGFRa mutation found in exon 12. The global “wild-type” cases, with no mutations in either the KIT or PDGFRa genes, were 13 (13.1%). The mean survival of those patients was approximately 46.9 (±43.9) months. Globally, 86.9% of Sardinian patients with GIST had a KIT or PDGFRa mutation; the former were more frequent in comparison with other Italian cohorts, while PDGFRa mutations were rare. No statistical differences in survival between mutated and wild-type cases, and between KIT and PDGFRa mutated cases were detected in our study.

Primary mucosal melanomas (MMs) are uncommon tumors originating from melanocytes located in the mucous membranes at various anatomic sites within the body. MM significantly differs from cutaneous melanoma (CM) regarding epidemiology, genetic profile, clinical presentation, and response to therapies. Despite these differences, that have important implications for both disease diagnosis and prognosis, MMs are usually treated in the same way as CM but exhibit a lower response rate to immunotherapy leading to a poorer survival rate. Furthermore, a high inter-patient variability can be observed in relation to therapeutic response. Recently, novel “omics” techniques have evidenced that MM lesions have different genomic, molecular, and metabolic landscapes as compared with CM lesions, thus explaining the heterogeneity of the response. Such specific molecular aspects might be useful to identify new biomarkers aimed at improving the diagnosis and selection of MM patients who could benefit from immunotherapy or targeted therapy. In this review, we have focused on relevant molecular and clinical advancements for the different MM subtypes in order to describe the updated knowledge relating to main diagnostic, clinical, and therapeutic implications as well as to provide hints on likely future directions.

Background: Gliomas are among the most challenging pathologies for neurosurgeons due to their infiltrative and recurrent nature in functionally relevant regions. Current knowledge confirms that gross total resection highly influence survival in patient with glioma. However, surgery performed in eloquent brain area, could seriously compromise the quality of life in patient with reduced life expectancy even more if it concerns the language function. Methods: 18 right-handed patients with perisylvian gliomas on the left hemisphere were prospectively analyzed over a period of 12 months. Standardized preoperative Diffusion-Tensor-Imaging based tractography of the five main language Tracts (Arcuate Fasciculus, Frontal Aslant Tract, Inferior Fronto-Occipital Fasciculus, Inferior Longitudinal Fasciculus, Uncinate Fasciculus) was navigated during the surgical procedure. Using a validated method, correlations were made between the pre-operative fascicles and their possible infiltration and surgical damage. The language status was assessed using the Aachen Aphasia Test. Results: In all nine patients who developed a permanent disorder there was pre-operative involvement of at least one fascicle and resection of at least one of these. In this way, areas of high risk of permanent language damage have emerged as a result of surgical injury: the temporoparietal junction, the middle portion of the FAT and the temporal stem. Conclusions: Navigated tractography has proven to be a user-friendly tool that can assess perioperative risk, guide surgical resection, and help the neurosurgeon to find that balance between tumor resection and function preservation.

Lung cancer is one of the most common and lethal cancers worldwide. Numerous medications targeting specific molecular alterations in non-small cell lung cancer have been introduced in the last decade and have revolutionized the clinical management of the disease. Their use has brought to a parallel evolution of molecular testing techniques to identify alterations in druggable molecular targets within the genetic material of the tumors. To perform molecular testing, biopsy or surgery tissue specimens are needed, which in addition allow the histological characterization of the tumors. Unfortunately, in real-life practice not all the patients are suitable for biopsy or surgery procedures. The use of liquid biopsy for blood extracted tumoral DNA analysis is a promising approach in unbiopsied cases, but it is also weighted by several methodological and technical limitations. We report here a case of histologically undiagnosed lung cancer managed with a liquid biopsy and subsequently with anti-EGFR treatment. Our report highlights that the use of liquid biopsy molecular testing in specific clinical situations can offer treatment opportunities for fragile patients affected by lung cancer.

The cure rate of germ cell tumours (GCTs) has significantly increased from the late 1970s since the introduction of cisplatin-based therapy, which to date remains the milestone for GCTs treatment. The exquisite cisplatin sensitivity has been mainly explained by the over-expression in GCTs of wild-type TP53 protein and the lack of TP53 somatic mutations; however, several other mechanisms seem to be involved, many of which remain still elusive. The findings about the role of TP53 in platinum-sensitivity and resistance, as well as the reported evidence of second cancers (SCs) in GCT patients treated only with surgery, suggesting a spectrum of cancer predisposing syndromes, highlight the need for a deepened understanding of the role of TP53 in GCTs. In the following report we explore the complex role of TP53 in GCTs cisplatin-sensitivity and resistance mechanisms, passing through several recent genomic studies, as well as its role in GCT patients with SCs, going through our experience of Center of reference for both GCTs and cancer predisposing syndromes.

9535 Background: To investigate the best sequential strategy, we started the SECOMBIT study, a randomized three parallel arms phase 2 study (NCT02631447). We used the combination of encorafenib/binimetinib (E+B) as targeted therapy (T-T) and the combination of ipilimumab/nivolumab (I+N) as immunotherapy (I-O). We explored the two different sequences and the “sandwich” strategy with a short course of target therapy, switched by combo immunotherapy at the best response, and not at progression of disease. In our previous report we have observed a trend in favor of the arms where I-O was given as first, confirmed by the first report of the DreamSeq study, a phase III study which compared the two different sequences with T-T and I-O. Here we updated the study data with a subgroup analysis. Methods: From Nov 2016 to May 2019, 37 centers in 9 countries enrolled 251 patients with untreated, metastatic BRAFV600 melanoma. Patients were randomized to Arm A [E+B until PD, followed by I+N], or Arm B (I+N until PD, followed by E+B) or Arm C (E+B for 8 weeks, followed by I+N until PD, followed by E+B). The overall survival (OS) is the primary endpoint of the study. Secondary endpoints included total progression-free survival (tPFS), 2- and 3-years survival rate, best overall response rate, duration of response, biomarkers evaluation. Results: The study primary endpoint was met in each arm with at least 30 patients alive at 24 months. The median follow-up estimated with the reverse Kaplan-Meier method was 37.1 months (IQR: 32.8-46.4). The OS at 2 and 3 years was calculated in the three arms for all patients, and in the subgroups normal or elevate LDH level and < 3 or ≥ 3 metastatic sites. OS and tPFS rates at 2 and 3 years are shown in the table. Conclusions: With a 37.1 months median follow-up, 2 and 3-years OS as well as tPFS rates are higher in Arm B and C. In line with recent findings, the SECOMBIT results confirm a better trend in favor of Arm B and C treatment sequence. The analysis of the secondary endpoints is ongoing. Clinical trial information: NCT02631447. [Table: see text]

Background Open articular fractures are an important issue due to their increasing incidence, along with the rise in motorcycle accidents. Oncology experience with allograft in orthopedics may be extended to cases of traumatic bone loss. Clinical case A 37-year-old man with an open fracture of the distal femur and proximal tibia presented a massive bone loss of the lateral femoral condyle after a motorcycle accident. Following Damage Control Surgery, a considerable lateral femoral condyle allograft was used to restore the joint anatomy. Results After an intensive rehabilitation program and two subsequent arthroscopic arthrolyses, the patient resumed walking without aids and recovered from stiffness (R.O.M. 0-130°). An MRI performed at median follow-up, revealed a favorable integration of the allograft. Discussion Allografts are a reasonable reconstructive option for young individuals with massive bone loss. An appropriately sized allograft is essential to restore an acceptable function as seen in the herein case.

The improvement of the immunotherapeutic potential in most human cancers, including melanoma, requires the identification of increasingly detailed molecular features underlying the tumor immune responsiveness and acting as disease-associated biomarkers. In recent past years, the complexity of the immune landscape in cancer tissues is being steadily unveiled with a progressive better understanding of the plethora of actors playing in such a scenario, resulting in histopathology diversification, distinct molecular subtypes, and biological heterogeneity. Actually, it is widely recognized that the intracellular patterns of alterations in driver genes and loci may also concur to interfere with the homeostasis of the tumor microenvironment components, deeply affecting the immune response against the tumor. Among others, the different events linked to genetic instability—aneuploidy/somatic copy number alteration (SCNA) or microsatellite instability (MSI)—may exhibit opposite behaviors in terms of immune exclusion or responsiveness. In this review, we focused on both prevalence and impact of such different types of genetic instability in melanoma in order to evaluate whether their use as biomarkers in an integrated analysis of the molecular profile of such a malignancy may allow defining any potential predictive value for response/resistance to immunotherapy.

The posterior arch of the atlas is usually not considered one of the main stabilizers of the cranio-cervical junction, allowing surgeons to its removal when needed with a relative certainty to preserve the stability of the atlo-axial segment. However, these considerations do not reflect the importance to examine the integrity of the posterior arch in the whole biomechanics of the atlas. Authors like Gebauer and Panjabi revealed, respectively in experimental and clinical conditions, how the atlas responds to an axial loading force, proving that the whole atlas is involved into horizontal conversion of axial forces and providing evidence supporting the preservation of the posterior arch. Other authors evaluated the risk for anterior arch fracture following C1 laminectomy. In this technical note three different techniques of posterior atlas arch reconstruction after surgical iatrogenic disruption are presented, considering both neoplastic and degenerative disease.

The pathological, clinical, and therapeutic features of colorectal cancer (CRC) depend on its anatomical localization. We investigated possible associations between the red blood cell distribution width (RDW) and CRC localization. Two-hundred eighty-eight consecutive patients with CRC were retrospectively studied. Demographic, clinical, pathological and laboratory data were retrieved from clinical records and reports. Median RDW values were significantly higher in patients with right-sided CRC when compared to those with CRC in other localizations (16.2, IQR: 14.5–20.0 vs 13.8, IQR: 13.0–16.1, p

The development and progression of melanoma, like that of all other malignancies, are based on the progressive acquisition of alterations affecting specific genes and pathways regulating melanocytic cell functions. Such molecular changes are determining the biological and clinical behavior of the disease, in terms of pathogenesis, prognosis, and propensity or resistance to therapeutic responses. As consequence, a detailed classification of the intrinsic intratumor heterogeneity of melanoma may identify patients with different molecular subtypes. The aim of the present review is to provide an update about all current advancements toward the definition of the main molecular profiles associated with tumor cell proliferation, invasion, survival, and migration underlying the different clinical phenotypes in patients with melanoma. In this sense, next-generation sequencing (NGS) approaches are extremely useful in unveiling the mutation landscapes in driver genes and pathways. Putting together all recent data, the melanoma pathogenesis scenario appears to be highly more complicated than hypothesized before. A molecular classification strategy based on such in-depth knowledge is becoming mandatory in order to discriminate melanoma patients for their clinical behavior and outcome. In other words, a more extensive molecular classification will increasingly be an integral part of the management of patients with melanoma.

Simple Summary The recent WHO classification of melanocytic tumors requires the implementation of combined phenotypic–genotypic diagnostics. For rare tumors, such as atypical deep penetrating nevi, there is insufficient information regarding genetic status, and it is not yet clear whether the observed unusual morphological cyto-architectures reflect a distinct genomic profile or are associated with an increased metastatic potential and aggressive clinical behavior. We report herein a comprehensive next-generation sequencing (NGS) analysis of a series of atypical DPNs, showing their mutational profile with some specific signatures for these rare and diagnostically challenging tumors. Abstract Deep penetrating nevi (DPNs) are rare melanocytic neoplasms consisting of pigmented spindled or epithelioid melanocytes with a distinctive wedge-shaped configuration showing activation of the WNT pathway, with unusual cyto-architectural features. It is unclear whether they show a distinct genomic profile associated with a diverse metastatic potential. We describe herein a cohort of 21 atypical DPNs analyzed by next-generation sequencing using the Ion AmpliSeq™ Comprehensive Cancer Panel. We found that β-catenin exon 3 was mutated in 95% and MAP kinase pathway genes in 71% of the cases. Less frequent mutations were observed in HRAS (19%) and MAP2K1 (24%). Isocitrate dehydrogenases 1 (IDH1) mutations, including R132C, V178I, and S278L, were identified in 38% of cases and co-existed with BRAF/HRAS mutations. The only case with progressive nodal disease carried alterations in the β-catenin pathway and mutations in IDH1 and NRAS (codon 61). By a comprehensive mutation analysis, we found low genetic heterogeneity and a lack of significant associations between specific gene mutations and histopathological features, despite atypical features. Whether the acquisition of an NRAS or IDH1 mutation in an atypical DPN may represent a molecular evolution implying a pathway to melanoma progression should be confirmed in a larger series.

Lung cancer is the leading cause of death for malignancy worldwide. Its molecular profiling has enriched our understanding of cancer initiation and progression and has become fundamental to provide guidance on treatment with targeted therapies. Testing the presence of driver mutations in specific genes in lung tumors has thus radically changed the clinical management and outcomes of the disease. Numerous studies performed with traditional sequencing methods have investigated the occurrence of such mutations in lung cancer, and new insights regarding their frequency and clinical significance are continuously provided with the use of last generation sequencing technologies. In this review, we discuss the molecular epidemiology of the main druggable genetic alterations in non-small cell lung cancer, namely EGFR, KRAS, BRAF, MET, and HER2 mutations or amplification, as well as ALK and ROS1 fusions. Furthermore, we investigated the predictive impact of these alterations on the outcomes of modern targeted therapies, their global prognostic significance, and their mutual interaction in cases of co-occurrence.

Summary Background The NIBIT-MESO-1 study demonstrated the efficacy and safety of tremelimumab combined with durvalumab in patient with unresectable mesothelioma followed up for a median of 52 months [IQR 49–53]. Here, we report 4-year survival and outcomes after retreatment, and the role of tumour mutational burden (TMB) in identifying patients who might have a better outcome in response to combined therapy. Methods NIBIT-MESO-1 was an open-label, non-randomised, phase 2 trial of patients with unresectable pleural or peritoneal mesothelioma who received intravenous tremelimumab (1 mg/kg bodyweight) and durvalumab (20 mg/kg bodyweight) every 4 weeks for four doses, followed by maintenance intravenous durvalumab at the same dose and schedule for nine doses. In this follow-up study, patients with disease progression following initial clinical benefit—ie, a partial repsonse or stable disease—were eligible for retreatment and with the same doses and schedules for tremelimumab and durvalumab as used in the NIBIT-MESO-1 trial. The primary endpoint, immune-related objective response rate, was evaluated per immune-related modified Response Evaluation Criteria in Solid Tumors (RECIST) or immune-related RECIST 1.1 criteria for patients with pleural or peritoneal malignant mesothelioma, respectively. Key secondary endpoints were overall survival and safety, and TMB was also evaluated post hoc in patients who had tumour tissue available before treatment. The intention-to-treat population was used for analysis of all efficacy endpoints. This study is registered with ClinicalTrials.gov , number NCT02588131 . Findings 40 patients were enrolled in the NIBIT-MESO-1 study between Oct 30, 2015, and Oct 12, 2016. At data cut-off, April 30, 2020, five (13%) of 40 patients were alive, and 35 (88%) patients had died of progressive disease. At a median follow-up of 52 months (IQR 49–53), median overall survival was 16·5 months (95% CI 13·7–19·2). Survival was 20% (eight of 40 patients) at 36 months and 15% (six of 40 patients) and 48 months. 17 (43%) of 40 patients met the criteria for enrolment in the retreatment study and were retreated with at least one dose of tremelimumab and durvalumab. No immune-related objective responses were observed in the 17 retreated patients. Seven (41%) of 17 patients achieved immune-related stable disease. From the start of retreatment to a median follow-up of 24 months (22·0–25·0), median overall survival was 12·5 months (95% CI 0·0–25·8), and survival at 12 months was 52·9%, at 18 months was 35·3%, and at 24 months was 23·5%. There were no grade 3–4 immune-related adverse events in the retreatment cohort. In a post-hoc analysis of 28 patients for whom tumour tissue before treatment was available, patients with a TMB higher than the median value of 8·3 mutations per Mb had a higher median overall survival compared with patients with TMB below the median value, but this difference was non-significant. Moreover, when patients were additionally stratified for ICI retreatment (n=13), there was a significant difference in survival between those with a TMB higher than the median of 8·3 mutations per Mb and those with TMB lower than the median in the retreated cohort (41·3 months vs 17·4 months; p=0·02). Interpretation Tremelimumab combined with durvalumab was associated with long-term survival in patients with mesothelioma. Retreatment was safe and resulted in clinically meaningful outcomes, thus suggesting its potential application in the clinical practice of mesothalioma patients. Funding NIBIT Foundation, Fondazione AIRC, AstraZeneca.

Background: Cutaneous malignant melanoma (CMM) is one of the most aggressive types of skin cancer. Currently, innovative approaches such as target therapies and immunotherapies have been introduced in clinical practice for the treatment of metastatic CMM. Data of clinical trials and real life studies that evaluate the outcomes of these therapeutic associations are necessary to establish their clinical utility. The aim of this study is to investigate the types of oncological treatments employed in the real-life clinical management of patients with advanced CMM in several Italian centers which are part of the Clinical National Melanoma Registry (CNMR), and the oncological outcomes obtained. Methods: CNMR collects data of patients with a histologically confirmed diagnosis of primary CMM treated in one of the 38 Italian institutions (hospitals, research institutes, etc.) participating in the network. Melanoma-specific survival and Overall survival were calculated. Kaplan-Meier curves and medians of OS and 95% CI are presented overall and by immunotherapy and target treatments. The Log-rank test compared curves by treatments. Multivariate Cox regression models were used to estimate the hazard ratios adjusting for confounders and other prognostic factors. Results: The median follow-up time was 36 months (range 1.2-185.1). 787 CMM were included in the analysis with completed information about therapies.Global immunotherapy showed a significant improved survival compared with all other therapies (p=0.001). 75% was the highest reduction of death reached by nivolumab/pembrolizumab immunotherapy (anti-PD1 HR=0.25 95% CI 0.14-0.42), globally immunotherapy was significantly associated with improved survival, either for anti-CTL A4 monotherapy or combined with anti-PD1 (HR=0.47;95% CI 0.33-0.66 and HR=0.26; 95% CI 0.15-0.46, respectively). Conclusions: The nivolumab/pembrolizumab and the combination of ipilimumab can be considered the best therapy to improve survival in a real-world-population. The CNMR can complement clinical registries with the intent of improving cancer management and standardizing cancer treatment.

Background and Objectives: The respiratory apparatus, generally affected by highly aggressive tumors like lung cancer and mesothelioma, is rarely affected by primary malignant melanoma. The aim of this review was to identify cases of primary malignant melanoma of the lung (PMML) published in the modern scientific literature, and to describe their main clinical, pathological and therapeutic features. Materials and Methods: A systematic search of publications in the electronic database PubMed has been performed using keywords, and the references of the selected articles were checked to identify additional missing studies. Results: Globally 52 papers reporting on 76 cases were identified. Among them there were 47 reports of a single case, three papers reporting on two cases each, and two larger case series published in 1997 and 2005 including eight and 15 cases, respectively. Conclusions: PMML was generally diagnosed in middle-aged males, without any apparent correlation with cigarette smoking. It was more frequently found in the lower lobes and the left lung. The tumors were generally pigmented, composed by epithelial and/or spindle cells with large nuclei and prominent nucleoli, nuclear atypia, and numerous mitotic figures; they commonly showed immunostaining for S-100, HMB 45 and Melan-A. Early detection and surgical resection were the main determinants of survival from this rare malignancy.

The rapid recovery of smell and taste functions in COVID-19 patients could be attributed to a decrease in interleukin-6 levels rather than central nervous system ischemic injury or viral damage to neuronal cells. To correlate interleukin-6 levels in COVID-19 patients with olfactory or gustatory dysfunctions and to investigate the role of IL-6 in the onset of these disorders, this observational study investigated 67 COVID-19 patients with taste or smell disorders or both, who did not require intensive care admission, admitted at COVID Hospital of Policlinico of Bari from March to May 2020. Interleukin-6 was assayed in COVID-19 patients with taste or smell disturbances at the time of admission and at the time of swab negativization. At the same time, patients have been given a specific survey to evaluate the severity of taste and smell disturbances. Of 125 patients with smell or taste dysfunctions at onset of disease, 67 fulfilled the inclusion criteria, while 58 were excluded because 35 of them required intensive care admission, 5 were unable to answer, 5 died, 7 had finished chemotherapy recently, and 5 refused to participate. The evaluation of taste and smell disorders was carried out using a survey performed at the time of admission and at the time of swab negativization. Sinonasal outcome test 22 (SNOT-22) was used as a reference for olfactory function assessment, and Taste and Smell Questionnaire Section of the US NHANES 2011-2014 protocol (CDC 2013b) was used as reference for gustatory function assessment. A venous blood sample was taken for each patient to measure IL-6 levels upon entry and at swab negativization. Interleukin-6 levels in COVID-19 patients in relation to olfactory or gustatory disorders were correlated from the time of their admission to the time of swab negativization. Statistically significant correlations were obtained between the decrease of interleukin-6 levels and the improvement of smell (p value < 0.05) and taste (p = 0.047) functions at swab negativization. The acquired results demonstrate the key role of interleukin-6 in the pathogenesis of chemosensitive disorders in COVID-19 patients.

Highlights • Separation surgery is a new concept for metastatic spinal cord compression treatment. • Stereotactic radiosurgery increased local control, overcoming radio-resistance’s idea. • The surgery goal shifted towards creating targets for radiations avoiding cord damages. • Minimal invasive strategies could allow quick return to systemic therapies., Introduction The new concept of separation surgery has changed the surgical paradigms for the treatment of metastatic epidural spinal cord compression (MESCC), shifting from aggressive cytoreductive surgery towards less invasive surgery with the aim to achieve circumferential separation of the spinal cord and create a safe target for high dose Stereotactic Body Radiation Therapy (SBRT), which turned out to be the real game-changer for disease’s local control. Discussion In this review a qualitative analysis of the English literature has been performed according to the rating of evidence, with the aim to underline the increasingly role of the concept of separation surgery in MESCC treatment. A review of the main steps in the evolution of both radiotherapy and surgery fields have been described, highlighting the important results deriving from their integration. Conclusion Compared with more aggressive surgical approaches, the concept of separation surgery together with the advancements of radiotherapy and the use of SBRT for the treatment of MESCC showed promising results in order to achieve a valuable local control while reducing surgical related morbidities and complications.

Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation status plays a predominant role in the management of MM patients. The aim of this study was to compare BRAF mutational testing performed by conventional nucleotide sequencing approaches with either real-time polymerase chain reaction (rtPCR) or next-generation sequencing (NGS) assays in a real-life, hospital-based series of advanced MM patients. Consecutive patients with AJCC (American Joint Committee on Cancer) stage IIIC and IV MM from Sardinia, Italy, who were referred for molecular testing, were enrolled into the study. Initial screening was performed to assess the mutational status of the BRAF and NRAS genes, using the conventional methodologies recognized by the nationwide guidelines, at the time of the molecular classification, required by clinicians: at the beginning, Sanger-based sequencing (SS) and, after, pyrosequencing. The present study was then focused on BRAF mutation detecting approaches only. BRAF wild-type cases with available tissue and adequate DNA were further tested with rtPCR (Idylla&trade, ) and NGS assays. Globally, 319 patients were included in the study, pathogenic BRAF mutations were found in 144 (45.1%) cases examined with initial screening. The rtPCR detected 11 (16.2%) and 3 (4.8%) additional BRAF mutations after SS and pyrosequencing, respectively. NGS detected one additional BRAF-mutated case (2.1%) among 48 wild-type cases previously tested with pyrosequencing and rtPCR. Our study evidenced that rtPCR and NGS were able to detect additional BRAF mutant cases in comparison with conventional sequencing methods, therefore, we argue for the preferential utilization of the aforementioned assays (NGS and rtPCR) in clinical practice, to eradicate false-negative cases and improve the accuracy of BRAF detection.

Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation status plays a predominant role in the management of MM patients. The aim of this study was to compare BRAF mutational testing performed by conventional nucleotide sequencing approaches with either real-time polymerase chain reaction (rtPCR) or next-generation sequencing (NGS) assays in a real-life, hospital-based series of advanced MM patients. Consecutive patients with AJCC (American Joint Committee on Cancer) stage IIIC and IV MM from Sardinia, Italy, who were referred for molecular testing, were enrolled into the study. Initial screening was performed to assess the mutational status of the BRAF and NRAS genes, using the conventional methodologies recognized by the nationwide guidelines, at the time of the molecular classification, required by clinicians: at the beginning, Sanger-based sequencing (SS) and, after, pyrosequencing. The present study was then focused on BRAF mutation detecting approaches only. BRAF wild-type cases with available tissue and adequate DNA were further tested with rtPCR (Idylla™) and NGS assays. Globally, 319 patients were included in the study; pathogenic BRAF mutations were found in 144 (45.1%) cases examined with initial screening. The rtPCR detected 11 (16.2%) and 3 (4.8%) additional BRAF mutations after SS and pyrosequencing, respectively. NGS detected one additional BRAF-mutated case (2.1%) among 48 wild-type cases previously tested with pyrosequencing and rtPCR. Our study evidenced that rtPCR and NGS were able to detect additional BRAF mutant cases in comparison with conventional sequencing methods; therefore, we argue for the preferential utilization of the aforementioned assays (NGS and rtPCR) in clinical practice, to eradicate false-negative cases and improve the accuracy of BRAF detection.

Idiopathic pulmonary fibrosis (IPF) is an aggressive pulmonary disease which shares several molecular, pathophysiological and clinical aspects with lung cancer, including high mortality rates. The antifibrotic drugs Nintedanib and Pirfenidone have recently been introduced in clinical practice for the treatment of IPF. Nintedanib is also used for the treatment of several malignancies, including non-small cell lung cancer (NSCLC) in combination with Docetaxel, while Pirfenidone showed some anti-neoplastic effects in preclinical studies. On the other hand, novel targeted agents and immunotherapies have been introduced in the last decade for the treatment of NSCLC, and some of them showed anti-fibrotic properties in recent studies. These evidences, based on the common pathophysiological backgrounds of IPF and lung cancer, make possible the mutual or combined use of anti-fibrotic and anti-neoplastic drugs to treat these highly lethal diseases. The aim of the present review is to depict the current scientific landscape regarding the repurposing of anti-neoplastic drugs in IPF and anti-fibrotic drugs in lung cancer, and to identify future research perspectives on the topic.

The objective of this population-based study is to describe epidemiological and genetic features of breast and ovarian cancer in North Sardinia, Italy. Patients who carry a high-risk mutation in one or both of the BRCA genes (BRCA1 or BRCA2) have a significantly increased risk of developing breast/ovarian cancer (BOC) and other cancers (e.g., prostate cancer in male). Epidemiological data on incidence distribution of breast/ovarian cancer from 2016 to 2019 in North Sardinia are obtained from the local tumor registry and from the cumulative results of 209 genetic testing for BRCA gene mutations performed in all young breast cancer patients and all women (over 50 years) with family history of BOC (total of 164 cases); further, 45 genetic testing is performed, on ovarian cancer patients, at any age. The results provide a different distribution of fraction mutations carried by women and a higher prevalence of the BRCA2 mutation in the north of Sardinia than the entire population and highlight the presence of specific germline mutation associated with the "founder effect" in distinct genetic subgroups reflecting genetic drift. Advances in next-generation sequencing technology, data analysis, and clinical investigation have revolutionized efforts to identify potential targets for BRCA molecular-based therapeutic agents.

The presence of ulceration has been recognized as an adverse prognostic factor in primary cutaneous melanoma (PCM).To investigate whether the extent of ulceration (EoU) predicts relapse-free survival (RFS) and overall survival (OS) in PCM.We retrieved data for 477 patients with ulcerated PCM from databases of the Italian Melanoma Intergroup. Univariate and multivariable Cox proportional hazard models were used to assess the independent prognostic impact of EoU.A significant interaction emerged between Breslow thickness (BT) and EoU, considering both RFS (P0·0001) and OS (P = 0·0006). At multivariable analysis, a significant negative impact of EoU on RFS [hazard ratio (HR) (1-mm increase) 1·26, 95% confidence interval (CI) 1·08-1·48, P = 0·0047] and OS [HR (1-mm increase) 1·25, 95% CI 1·05-1·48, P = 0·0120] was found in patients with BT ≤ 2 mm, after adjusting for BT, age, tumour-infiltrating lymphocytes, sentinel lymph node status and mitotic rate. No impact of EoU was found in patients with 2·01-4 mm and4 mm BT.This study demonstrates that EoU has an independent prognostic impact in PCM and should be recorded as a required element in pathology reports.

Background: Next-generation sequencing (NGS) needs to be validated and standardized to ensure that cancer patients are reliably selected for target treatments. In Italy, NGS is performed in several institutions and harmonization of wet and dry procedures is needed. To this end, a consortium of five different laboratories, covering the most part of the Italian peninsula, was constituted. A narrow gene panel (SiRe®) covering 568 clinically relevant mutations in six different genes (EGFR, KRAS, NRAS, BRAF, cKIT, and PDGFRα) with a predictive role for therapy selection in non-small cell lung cancer (NSCLC), gastrointestinal stromal tumor, colorectal carcinoma (CRC), and melanoma was evaluated in each participating laboratory. Methods: To assess the NGS inter-laboratory concordance, the SiRe® panel, with a related kit and protocol for library preparation, was used in each center to analyze a common set of 20 NSCLC and CRC routine samples. Concordance rate, in terms of mutation detected and relative allelic frequencies, was assessed. Then, each institution prospectively analyzed an additional set of 40 routine samples (for a total of 160 specimens) to assess the reproducibility of the NGS run parameters in each institution. Results: An inter-laboratory agreement of 100% was reached in analyzing the data obtained from the 20 common sample sets; the concordance rate of allelic frequencies distribution was 0.989. The prospective analysis of the run metric parameters obtained by each center locally showed that the analytical performance of the SiRe® panel in the different institutions was highly reproducible. Conclusions: The SiRe® panel represents a robust diagnostic tool to harmonize the NGS procedure in different Italian laboratories.

Il carcinoma cutaneo a cellule squamose è il secondo tumore della pelle per incidenza, ma ancora poco conosciuto. L'A.I.Ma.Me (Associazione Italiana Malati di Melanoma e Tumori della Pelle), a partire dal 2019, ha dato vita a un Gruppo di lavoro specifico sul Carcinoma Cutaneo a Cellule Squamose (CSCC) e, per sensibilizzare tutti i cittadini sulle regole di prevenzione e dar voce ai pazienti, ha realizzato il Libro Bianco su questa patologia. Il presente capitolo introduttivo affronta le caratteristiche di questo tumore partendo dalla descrizione delle caratteristiche dei tumori cutanei: melanoma e tumori non-melanoma. Vengono poi descritte le caratteristiche del CSCC, le modalità di sviluppo, i fattori di rischio, i sottotipi istologici del CSCC, i fattori prognostici per recidiva/avanzamento di malattia. I meccanismi molecolari alla base del CSCC sono scarsamente chiariti e i driver della progressione della malattia non sono stati completamente definiti e vengono prese inconsiderazione le attuali conoscenze in questo campo.

Spontaneous regressions of calcified thoracic disc herniations (TDH) have been rarely described. Since now, no previous papers hypothesized that radiological factors could be able to predict the evolution of the herniation. This study shows that the radiodensity of the herniated material in herniations with spontaneous resolution might differ from that of operated herniations. This is a retrospective study. A cohort of patients that underwent surgical removal of a calcified thoracic disc herniation has been compared to two cases conservatively treated and a case with spontaneous regression. CT scans were evaluated to compute the average radiodensity [in Hounsfield units (HU)] of the calcified components of the herniations. All the slices of the CT scans involving the calcification were evaluated in their axial images and in their coronal and sagittal reformations. Comparison of the slices radiodensity (spontaneously resolved versus surgically treated) was performed with the unpaired Student’s t test. The mean density of calcification in the case with spontaneous regression was found to be 454 HU. In the control group of surgical cases observed, and the conservatively treated cases, the mean HU density showed higher values (respectively, 827 and 853 HU) (p

BackgroundThe coronavirus disease 2019 (COVID-19) pandemic has overwhelmed the health systems worldwide. Data regarding the impact of COVID-19 on cancer patients (CPs) undergoing or candidate for immune checkpoint inhibitors (ICIs) are lacking. We depicted the practice and adaptations in the management of patients with solid tumors eligible or receiving ICIs during the COVID-19 pandemic, with a special focus on Campania region.MethodsThis survey (25 questions), promoted by the young section of SCITO (Società Campana di ImmunoTerapia Oncologica) Group, was circulated among Italian young oncologists practicing in regions variously affected by the pandemic: high (group 1), medium (group 2) and low (group 3) prevalence of SARS-CoV-2–positive patients. For Campania region, the physician responders were split into those working in cancer centers (CC), university hospitals (UH) and general hospitals (GH). Percentages of agreement, among High (H) versus Medium (M) and versus Low (L) group for Italy and among CC, UH and GH for Campania region, were compared by using Fisher’s exact tests for dichotomous answers and χ2 test for trends relative to the questions with 3 or more options.ResultsThis is the first Italian study to investigate the COVID-19 impact on cancer immunotherapy, unique in its type and very clear in the results. The COVID-19 pandemic seemed not to affect the standard practice in the prescription and delivery of ICIs in Italy. Telemedicine was widely used. There was high consensus to interrupt immunotherapy in SARS-CoV-2–positive patients and to adopt ICIs with longer schedule interval. The majority of the responders tended not to delay the start of ICIs; there were no changes in supportive treatments, but some of the physicians opted for delaying surgeries (if part of patients’ planned treatment approach). The results from responders in Campania did not differ significantly from the national ones.ConclusionOur study highlights the efforts of Italian oncologists to maintain high standards of care for CPs treated with ICIs, regardless the regional prevalence of COVID-19, suggesting the adoption of similar solutions. Research on patients treated with ICIs and experiencing COVID-19 will clarify the safety profile to continue the treatments, thus informing on the most appropriate clinical conducts.

Our group is long evaluating epidemiological and genetic features of breast-ovarian cancers and other BRCA-related tumor in North Sardinia, Italy. Individuals carrying a predisposing mutation in one or both of the BRCA (BRCA1 or BRCA2) genes have a significantly increased risk of developing breast/ovarian cancer (BOC) and other BRCA-related tumors such as pancreatic and prostate cancer. Epidemiological data on incidence and distribution of such cancers are obtained from the local tumor registry; 411 genetic testing for BRCA gene mutations have been performed. Germline NGS-based analysis were performed in all young breast cancer patients or all women (over 50 years) with a strong family history of BOC (total of 384 cases), males with sporadic breast cancer (6 cases) and one with prostate cancer, and lately patients with pancreatic cancer (20 cases). Genetic testing for breast and ovarian cancer has clinical importance for identification of potentially affected families and for cancer prevention. Approximately, 5-10% of breast and ovarian cancer are familiar while about 90% are sporadic tumor cases. BRCA germline mutations account for approximately 10-15% of BOC cancers. Confirming previous results from our group, a higher prevalence of the BRCA2 mutation was observed in North Sardinia as compared to the general population and highlight the presence of specific germline mutation associated with the "founder effect" in distinct genetic subgroups reflecting genetic drift. Conversely, BRCA1 mutations are much more strongly associated at higher rates of BOC cancer in South Sardinia. Heterozygous BRCA mutations determine loss of function of the remaining wild-type allele, resulting in deficient homologous recombination DNA repair, which causes genetic aberrations that drive carcinogenesis. PARP inhibition in mutated tumor cells with deficient homologous-recombination repair generates unrepaired DNA single-strand breaks that are likely to cause the accumulation of DNA double-strand breaks and collapsed replication forks. Currently, analysis performed on somatic tissue allows to identify a greater number of mutations in BRCA-related tumor (additional 5-7% of cases as compared to germline testing). As consequence, the current mutational analysis is based on the use of multigenic NGS customized "Oncomine Cancer panels" that allow to analyze additional genes involved in the onset of cancer (SMAD4, ATM, CHEK2, APC, NF, KRAS, CDKN2A) as well as in maintaining the genome integrity (POLE, PALB2, PTEN, TP53, PIK3CA). Notably, detection of three PIK3CA hot spot mutations - E545K, E542K, H1047R (detected in about 40% cases) - is a new targeted therapy with PIK3CA inhibitors and represent a new strategy to overcome resistance in mutated advanced breast cancer. Analogously, combination of PARP and AR (androgen receptor) inhibitors was demonstrated to exert a benefit into the treatment of prostate cancer. The androgen receptor promotes DNA damage repair and upregulates PARP-mediated repair pathways, thus increasing the effectiveness of the PARP inhibitor. Finally, in pancreatic adenocarcinoma, the presence of mutations in cancer susceptibility genes and the involvement of external factors (such as those underlying local inflammation) play a role in pathogenesis along with the occurrence of somatic mutation in KRAS and other "driver" genes (SMAD4 and CDKN2A), leading to the use of new targeted therapies.

Circulating tumor DNA (ctDNA), commonly known as liquid biopsy, is shed from tumor cells into the bloodstream of the patient. ctDNA containing somatic mutations which occur in tumor tissues may represent valid biomarkers to follow disease progression. Despite the major challenge remains assay sensitivity and specificity for ctDNA analysis, advancements represented by the use of new technical approaches such as NGS and droplet digital PCR assays - able to detect and quantify small amounts of mutated DNA molecules - have enabled the use of mutations as predictive/prognostic tool in the clinical practice. Several studies in melanoma, breast, lung and colorectal cancers demonstrated the potential clinical application of ctDNA analysis at each stage of cancer management: early diagnosis, molecular profiling for prognosis, detection of residual disease, monitoring response or resistance to treatment. In malignant melanoma and breast cancer, recent studies reveal that ctDNA could potentially be an excellent marker of residual disease, guiding therapeutic strategies. Moreover, occurrence of circulating KRAS mutations may predict failure of anti-EGFR therapies or, on the contrary, their disappearance from blood may address the rechallenge with EGFR inhibitors in patients with colorectal cancer (CRC). In our preliminary experience within the ongoing clinical trial DISTINCTIVE (Second-D line folfiri/aflIbercept in proSpecTIvely stratified, anti-EGFR resistaNt, metastatic coloreCTal cancer patients wIth RAS Validated wild typE status), Sardinian patients (N=44) with RAS wild-type advanced CRC who received first-line anti-EGFR treatment were investigated for circulating RAS or BRAF mutations at disease progression in order to determine whether the status of tumor biomarkers changes during tumor evolution. We found that about one third of patients presented a circulating oncogene mutation (29.5% in KRAS, 4.5% in NRAS, and 2.3% in BRAF), whereas in the remaining larger fraction (>60%) of the series the lack of mutated ctDNA addressed cases to the rechallenge therapy with anti-EGFR drugs. Efforts to improve the outcome of the oncological treatments for non-small cell lung cancer (NSCLC), together with the technological advances in DNA sequencing, led to the development of new therapeutic strategies. Subsets of patients with adenocarcinoma and activating mutations within the kinase domain of the epidermal growth factor receptor (EGFR) gene have been successfully treated with selective tyrosine kinase inhibitors (TKI). We previously described the molecular epidemiology of EGFR, KRAS, BRAF, ALK and MET genetic alterations and their correlations with the demographic and clinical characteristics of 1,440 Sardinian patients with lung adenocarcinoma. In NSCLC patients treated with the EGFR TKIs, the T790M mutation in EGFR gene represents the most common mechanism of acquired resistance. We started the analysis of 165 consecutive liquid biopsies from advanced NSCLC patients treated with TKIs, detecting the T790M in 11.5% of cases. EGFR MutationPositive (%)Negative (%) T790M 19 (11.5%) 146 (88.5%) Exon 19 Deletions 35 (21.2%) 130 (78.8%) L858R 18 (10.9%) 147 (89.1%) In addition to complete such clinical trials, our group will extend the analysis of ctDNAs in other neoplastic pathologies through the participation to further studies by using NGS-based strategies.

Idiopathic Pulmonary Fibrosis (IPF) is a specific, chronic and progressive form of interstitial pneumonia with sever prognosis and poor survival rates (2/3 years from diagnosis). IPF incidence is estimated in 2/30 cases per 100.000 persons/years, mainly affecting over 65 years old individuals. Although pathophysiological mechanisms are not fully understood, environmental and occupational factors, smoking, viral infections, and traction injury of the lung are recognized as causative elements leading to chronic damage to the alveolar epithelium. IPF is considered an epithelium-driven disease where recurring micro-injuries in lung epithelia promote an imbalance between profibrotic (TGF-?, PDGF, bFGF, IL-1, TNF-?) and anti-fibrotic (KGF, HGF, EPC) mediators. In this scenario, chronic fibrosis replaces normal repair mechanisms: excessive fibroblasts proliferation and aberrant extracellular matrix remodeling determine an irreversible distortion of lungs architecture and subsequent evolution in honeycombing areas. IPF shares several molecular, pathophysiological and clinical aspects with lung cancer, including high mortality rates; in particular, both diseases share genetic and epigenetic factors, enhanced proliferation and abnormal activation of specific signaling transduction pathways. The prevalence of lung cancer, particularly adenocarcinoma, in patients with IPF in clinical studies ranges from 31% in older reports to 10-17% in more recent reports. Nevertheless, these figures were consistently lower in epidemiological studies, ranging from 4% to 7%. IPF patients affected by lung cancer are commonly male, smokers or ex-smokers, at their 6th or 7th decade of life; lung cancer arises generally 2-3 years after the diagnosis of IPF, and affects mostly the peripheral aspect of the lower lobes of the lungs where fibrosis is more frequent and consistent Considering the above-mentioned similarities, we started a comparative study of the mutational landscape in IPF and lung cancer tissue biopsies. We have already collected formalin-fixed, paraffin embedded (FFPE) IPF tissues samples (28 cases over 49 recorded in our database), 15% of which has progressed to lung cancer (IPF/K). In addition, we aim to carry out further comparisons using as a reference sample a previously analyzed cohort of IPF-unrelated lung adenocarcinomas. Our study is structured in 3 steps: a) clinical and pathological data collection: comorbidity, lifestyle (e.g. smokers/non-smokers), respiratory parameters (PFT, FEV, FVC, TLC, DLCO, BPCO, chronic respiratory failure degree and pulmonary hypertension); b) histopathologic classification and centralized FFPE sections processing; c) DNA and RNA extraction from FFPE followed by NGS mutational analysis using a multi-genic panel (52 main oncogenes and tumor suppressor genes involved in tumorigenesis). Our preliminary results on 28 DNA/RNA from FFPE IPF tissue biopsies, we found 16 missense variants spreading on 11 genes. Among them, 3 are filtered out as pathogenic by prediction algorithms in FGFR3, MTOR and PIK3CA genes respectively, that play a pivotal role in cellular growth and proliferation mechanisms. Once completed the mutational analyses in all collected samples (IPF and IPF/K), we plan to evaluate correlation between different molecular profiles and clinical-pathological parameters in order to define potential patients subgroups. Furthermore, all results will be compared with the reference sample of lung adenocarcinoma previously processed using the the same panel. Defining consistent patients sub-groups is indeed crucial for better understanding the molecular mechanisms underlying the IPF pathogenesis and its progression to lung cancer.

Lung cancer is one of the most common malignancies worldwide. Despite a significant amount of basic and clinical research, mortality rates remain extremely high, especially for patients affected by advanced stage disease. Recently, new molecules playing several roles in the pathogenesis, diagnosis, and, potentially, clinical management of lung cancer are under investigation, including noncoding fragments of the human genome, also known as noncoding RNAs (ncRNAs). NcRNAs are commonly divided into two categories according to their size. The first category includes small ncRNAs, such as the recently discovered miRNAs, siRNAs, and the classical cellular RNAs (ribosomal, transfer, and other RNAs). Noncoding RNAs greater than 200 nucleotides represent a further category that includes long noncoding RNAs (lncRNAs). LncRNAs have numerous biological and pathophysiological effects. Numerous studies have recently investigated their involvement in the oncogenesis and the progression of pulmonary malignancies. In this chapter, we summarize the current knowledge regarding the role of lncRNAs in the pathogenesis, diagnosis, and clinical management of non-small cell lung cancer.

BACKGROUND: Little is known on whether melanocortin 1 receptor (MC1R) associated cutaneous melanoma (CM) risk varies depending on histological subtype and body site, and whether tumour thickness at diagnosis (the most important prognostic factor for CM patients) differs between MC1R variant carriers and wild-type individuals. OBJECTIVE: We studied the association between MC1R variants and CM risk by histological subtype, body site, and Breslow thickness, using the database of the M-SKIP project. METHODS: We pooled individual data from fifteen case-control studies conducted during 2005–2015 in Europe and the USA. Study-specific, multi-adjusted odds ratios were pooled into summary odds ratios (SOR) and 95% confidence intervals (CI) using random-effects models. RESULTS: 6891 CM cases and 5555 controls were included. CM risk was increased among MC1R variant carriers vs. wild-type individuals. The increase in risk was comparable across histological subtypes (SOR for any variant vs. wild-type ranged between 1.57 and 1.70, always statistical significant) except acral lentiginous melanoma, for which no association emerged; and slightly greater on chronically (1.74, 95% CI 1.47–2.07) than intermittently (1.55, 95% CI 1.34–1.78) sun-exposed skin. CM risk was greater for those carrying ‘R’ vs. ‘r’ variants; correlated with the number of variants; and was more evident among individuals not showing the red hair colour phenotype. Breslow thickness was not associated with MC1R status. CONCLUSION: MC1R variants were associated with an increased risk of CM of any histological subtype (except ALM) and occurring on both chronically and intermittently sun-exposed skin.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive human pancreatic cancer type accounting for about 90% of all malignant pancreatic neoplasms. It represents the fourth leading cause of cancer death in the Western World, projected to become the second cause by 2030. In our country, the PDAC incidence is about 14,000 new cases per year and represents 3% of all cancer cases (AIRTUM 2019), equally distributed between the sexes. Up to 15-20% of patients are eligible for resection at diagnosis. The 5 years overall survival for PDAC patients is around 8%; even for patients eligible for resection, the 5 years survival is no more than 15-20%. At least 25% of pancreatic cancers harbor "actionable" molecular alterations, defined as molecular alterations for which there is clinical evidence of a predictive benefit from a specific therapy (targeted therapy). We started a study by Next Generation Sequencing (NGS) analysis in primitive and metastatic PDAC tissues from the same patients, to define the molecular alterations pattern underlying neoplastic progression in pancreatic cancer. Tumor samples - 50 primitive PDAC cases and 50 related lymph node metastases - have been selected from archived formalin-fixed and paraffin-embedded (FFPE) samples, of which clinical-pathological data are available. To date have been analyzed 11 samples (4 primitive/metastasis matched cases and 3 primitive tumors) by NGS screening using "Oncomine Tumor specific panel", including main genes involved in onset and progression of PDAC (KRAS, CDKN2A, SMAD4, CHEK2, APC, NF1), in genome damage repair (POLE, PALB2, PTEN, TP53), and in mismatch repair mechanism (MLH1, MSH2.MSH3, MSH6). Preliminary data of mutational analysis are shown in the following Tables. The main endpoint for the analysis of all of cases is to identify mutational patterns to relate to clinical-pathological parameters of PDAC, as a starting point for finding novel tumor biomarkers associated with the disease.

Colorectal cancer (CRC) is the third and second most common malignancy in men and women, respectively. Approximately, one in four CRC patients is diagnosed with an advanced disease (stage IV, according to the American Joint Committee on Cancer/AJCC). Advanced CRC has a high mortality rate, consistently ranking in the top three causes of cancer-related deaths, whereas treatment success rates range from 70-90% in the localized disease. CRC is predominantly associated with developed countries and a Western lifestyle and diet. In CRC pathogenesis, somatic mutations in RAS and BRAF oncogenes play an important role and represent a key focus for the use of targeted therapies. In recent past years, the epidermal growth factor receptor (EGFR)-depending pathway has been largely exploited for personalized therapies and EGFR has become a key target of specific inhibitors to treat metastatic CRCs. Activating mutations in RAS are recognized as a strong predictor of resistance to EGFR-targeted agents, since they cause a constitutive phosphorylation of the RAS proteins - independent on activation status of the upstream EGFR protein - which in turn permanently promote cell proliferation and drastically reduce the effects of the EGFR inhibition. In particular, monoclonal antibodies targeting EGFR or the vascular endothelial growth factor (VEGF) are biologic-targeted agents, both of which have improved outcomes when utilized in combination with chemotherapy in metastatic CRC. In particular, EGFR inhibitors are included into the treatment of CRC with RAS and BRAF wild-type status, the anti-VEGF agent bevacizumab in RAS-mutated CRCs, and BRAF inhibitors in BRAF-mutant tumors. Moreover, occurrence of microsatellite instability (the MSI phenotype), which is present in

In the era of personalized medicine, BRAF mutational assessment is mandatory in advanced-stage melanoma and non-small cell lung cancer (NSCLC) patients. The identification of actionable mutations is crucial for the adequate management of these patients. To date various drugs have been implemented in clinical practice. Similarly, various methods may be adopted for the identification of BRAF mutations. Here, we briefly review the current literature on BRAF in melanoma and NSCLC, focusing attention in particular on the different methods and drugs adopted in these patients. In addition, an overview of the real-world practice in different Italian laboratories with high expertise in molecular predictive pathology testing is provided.