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Between January 2003 and January 2015, women at genetic risk of developing breast and/or ovarian cancer were selected for a surveillance program. The monitoring strategy consisted of the association of breast ultrasound (US), every six months, and Rx-mammography (XM) and breast MRI (MRI) to be performed annually. To date, 29 women have been included in the surveillance program: BRCA1 mutation carriers, 18; BRCA2 mutation carriers, 11. Eight women (28%) had already developed breast and/or ovarian cancer, while 21 (72%) were unaffected carriers. At a median surveillance time of 33 months (range 0-144), 4 incidental breast cancers were diagnosed in 4 BRCA1/2 mutation carriers (4/29, 14%), 2 BRCA1+ and 2 BRCA2+; 3/21 (14%) unaffected carriers and 1/8 (13%) previously affected carriers. Two cancers (50%) were detected by all three diagnostic tools; 2 (50%) were identified only by US and MRI in patients aging 33 and 43, respectively. Characteristics of patients with breast cancer diagnosed during surveillanceagemutated geneprevious cancerUSXMMRIhistologysurveillance time (months)33BRCA2-+-+ductal74+43BRCA2-+-+ductal40+45BRCA1-+++ductal60+40BRCA1yes+++ductal74+ One of these four patients, affected by breast cancer, died at 31 months of overall survival; three underwent surgery, chemotherapy and radiotherapy and are still disease free, in follow-up at 59, 28 and 26 months, respectively. In conclusion, 29 BRCA1/2 carriers have been included in the surveillance program, the median age was 42.5 months (range 27-68) and the median surveillance time was 33 months (range 0-144). Our preliminary data confirm the 3% expected rate of diagnosed cancers and the effectiveness of performing the triple diagnostic surveillance with US, XM and MRI. Citation Format: Sidoni T, Cocciolone V, Cannita K, Di Giacomo D, Ciccozzi A, Bafile A, Pizzorno L, Resta V, Marsecano C, Ferrari F, Di Cesare E, Ficorella C, Ricevuto E. Surveillance program for women carrying BRCA1/BRCA2 genetic predisposition. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-09-14.

Breast cancer (BC) is a common and heterogeneous disease, of which six molecular subtypes, characterized by different biological features and clinical outcomes, were described. The identification of additional biomarkers able to further connote and distinguish the different BC subtypes is essential to improve the diagnostic, prognostic and therapeutic strategies in BC patients. MicroRNAs (miRNAs) are short non-coding RNA involved in several physiological and pathological processes, including cancer development and progression. In particular, circulating miRNAs, which can be found in an adequately stable structure in serum/plasma of cancer patients, are emerging as very promising non-invasive biomarkers. Several studies have analyzed the potential role of circulating miRNAs as prognostic and therapeutic markers in BC. In the present review we describe circulating miRNAs, identified as putative biomarker in BC, with special reference to different BC molecular subtypes.

Lung cancer represents the major cause of cancer-related mortality in countries with a high degree of socioeconomic development, with non-small cell lung cancer (NSCLC) accounting for approximately 85% of all primary lung cancers (1,2).

The aim of the present study was to assess the expression of select DNA repair and synthesis genes in non-small-cell lung cancer (NSCLC) according to KRAS mutation status. ERCC1, TS, RRM1, and BRCA1 mRNA expression levels were assessed from either primary or metastatic tumor specimens of patients diagnosed with epidermal growth factor receptor (EGFR) wild-type (WT) advanced NSCLC. Total RNA was isolated from paraffin-embedded tumor specimens using the RNeasy FFPE kit and automatically purified using a QiaCube instrument. Quantification levels were analyzed by real-time one-step RT-PCR using QuantiFast technology, and the results were compared considering β-actin as the internal reference gene. One hundred and eighty-four patients with advanced NSCLC were evaluated for the analysis, of which 92 were KRAS-mutants. Nearly all patients had adenocarcinoma histology (96.7%). Among KRAS-mutants, the majority had a KRAS codon 12 mutation (88%), the most common being G12C (44.4% of cases). Mean ERCC1 levels were indicated to be significantly higher in KRAS-mutants when compared with KRAS WT patients (3,234±6.63 vs. 184±1.24; P=0.05). However, mean TS levels were significantly lower in the KRAS-mutant subgroup compared with the KRAS WT subgroup (4,481±3.756 vs. 5,941±6.4; P=0.039). KRAS-mutant NSCLCs are more likely to express high ERCC1 and low TS levels. This finding may suggest different sensitivity to cytotoxic chemotherapy according to KRAS mutation status.

Epidermal growth factor receptor (EGFR)-mutated (exons 18-21) advanced non-small cell lung cancers (NSCLCs) are generally characterized by exquisite sensitivity to treatment with an EGFR-tyrosine kinase inhibitor (-TKI). First-generation or reversible EGFR-TKIs include gefitinib and erlotinib, while, more recently, second-generation or irreversible EGFR-TKIs have been developed, namely afatinib and dacomitinib, with the aim of overcoming/delaying acquired resistance to treatment. Nevertheless, clinical trials have shown that resistance eventually emerges after a median time of slightly less than one year, regardless of whether first- or second-generation EGFR-TKIs are used. In this context, a secondary EGFR mutation in exon 20, namely T790M, has been found to be responsible for approximately 60% of cases of acquired resistance. Alternatively, T790M resistance mutation can be found de novo, in which case it limits the antitumor activity of both first- or second-generation EGFR-TKIs. Osimertinb is an orally bioavailable, third-generation EGFR-TKI that acts by irreversibly binding both EGFR activating mutations and T790M, while sparing wild-type EGFR. On this basis, osimertinib has proven more efficacious than platinum-based chemotherapy in the setting of EGFR T790M-positive NSCLCs pretreated with a first- or second-generation EGFR-TKI. More recently, in another phase 3 trial, osimertinib outperformed gefitinib or erlotinib as first-line treatment of EGFR-mutated (ex19del or L858R) advanced NSCLCs, thus emerging as a new standard of care in this setting. In the present review, we will discuss the preclinical and clinical development of osimertinib, briefly touching upon its activity in special populations and biomarkers of sensitivity to treatment.

Several cancers, especially non-small cell lung cancer (NSCLC), are able to escape the immunosurveillance of tumor-infiltrating lymphocytes (TILs); among the molecules involved, the indoleamine 2,3-dioxygenase 1 (IDO-1) and the programmed cell death ligand-1 (PD-L1) play a crucial role. These aspects are of great interest in the current immunotherapeutic era, therefore the current study analyses the TILs, IDO-1, and PD-L1 interactions and their correlations with clinicopathological parameters and prognosis in NSCLC. One hundred ninety-three NSCLC surgical specimens, formalin-fixed, and paraffin-embedded were assessed for TILs density, TILs localization, IDO-1 (clone 4.16H1), and PD-L1 (clone E1L3N) immunohistochemical expressions. This data was correlated with clinicopathological parameters, disease free, and overall survivals. IDO-1 and PD-L1 high expressions were related to the solid pattern of adenocarcinomas (respectively p = 0.036 and p = 0.026); high PD-L1 expression was correlated with squamous histotype (p = 0.048). IDO-1 overexpression correlated with former smokers (p = 0.041), higher adenocarcinoma stages (p = 0.039), and with both higher TILs density and PD-L1 expression (respectively p = 0.025 and p = 0.0003). A better prognosis was associated with TILs intratumoral or mixed localizations (p = 0.029). TILs localization affects NSCLC prognosis; the higher expression of IDO-1 and PD-L1 in poorly differentiated and more aggressive lung adenocarcinomas, as well as the correlation between high PD-L1 expression and squamous cell histotype, confirm the more efficient immunoescaping of these NSCLC subgroups.

The Feline Immunodeficiency Virus (FIV) is a lentivirus that affects cats worldwide. The detection ofthe virus’ 24 Kda capsid protein (p24) antibody is the most common way to diagnose FIV infections.In Brazil, the most used commercial assay for FIV diagnosis is a rapid ELISA test based on the lateralflow principle, with high sensitivity and specificity (SNAP®test). However, the high cost of this importedassay undermines not only the diagnosis of this infection but also its epidemiology. The objective of thisstudy was to optimize and validate an indirect ELISA using the protein p24 recombinant antigen (r-p24)with national technology and maintaining sensitivity and specificity comparable to those of availablecommercial rapid ELISA tests. Twenty-six reference cats’ sera samples (13 negative and 13 positives for FIV),previously analyzed by PCR and the SNAP® test, were used as a reference to optimize the ELISA. Serumsamples of 226 cats from private owners and from shelters of the metropolitan area of Rio de Janeirowere used to validate the assay. The sensitivity and specificity of the r-p24 ELISA were 95.4% and 99.4%,respectively. When compared to the SNAP® Combo Test, the accuracy of r-p24 ELISA was 99%, with aKappa index of 0.96. Our results indicate that the national technology-based ELISA r-p24 has sensibilityand specificity values that are comparable to rapid ELISA kits. This test is therefore recommendable, usingnational technology for FIV routine diagnosis, research, and epidemiological studies. O Vírus da Imunodeficiência Felina (FIV) é um lentivírus que afeta gatos em todo o mundo. A detecçãodo anticorpo para proteína do capsídeo viral de 24 kDa (p24) é a forma mais comum para diagnosticarinfecções pelo FIV. No Brasil, o ensaio comercial mais utilizado no diagnóstico FIV é um teste rápidode ELISA (SNAP®test), baseado no princípio de fluxo lateral, com alta sensibilidade e especificidade.No entanto, o alto custo deste ensaio importado inviabiliza não só o diagnóstico da infecção, como tambémsua epidemiologia. O objetivo deste trabalho foi aperfeiçoar e validar um teste de ELISA indireto, comtecnologia nacional, utilizando o antígeno recombinante da proteína p24 (r-p24), com sensibilidade eespecificidade comparáveis às dos testes rápidos, comercialmente disponíveis. Vinte e seis amostras desoro gatos (13 negativos e 13 positivos para FIV), previamente analisados por PCR e Snap®, foram utilizadascomo amostras de referência para aperfeiçoar o ELISA e 226 amostras de soro de gatos, de proprietáriosprivados ou de abrigos da área metropolitana Rio de Janeiro foram utilizadas para validar o ensaio. O ELISAr-p24 apresentou uma sensibilidade de 95,4% e 99,4% de especificidade quando comparadas às do testeSNAP Combo, uma precisão de 99% e um índice Kappa de 0.96. Nossos resultados indicam que o ELISAr-p24 apresentou sensibilidade e especificidade comparáveis às dos testes rápidos, utilizando tecnologianacional para diagnóstico, pesquisa e estudos epidemiológicos do FIV.

G2 ductal infiltrating carcinomas are a heterogeneous group of tumours with ambiguous clinical significance. This is because G2 carcinomas are almost always the largest category and poorly reproducible. Mitotic count (MC) is one of the causes of poor histological grading reproducibility. The phosphoistone H3 (PPH3) antibody improves identification of mitotic figures. The aim of our study is to demonstrate whether using a new histological grading system based on PPH3 immunostaining to assess MC can re-stratify G2 category. We selected 100 cases of G2 invasive carcinoma. The mitotic score was accurately reevaluated performing MC on PPH3 immunostained sections. 21/100 G2 cases (21%) showed the same mitotic score both with hematoxilin and eosin (H&E) and PPH3 while 79 cases (79%) with PPH3 shifted to a higher mitotic score. After re-grading the 100 G2 cases based on the assessment of mitotic score with PPH3 only 53 cases (53%) were confirmed as G2, while 47 cases (47%) had shifted to G3. Finally we reclassified early tumours in the surrogate molecular subtype according to the 2013 St. Gallen Conference criteria and found that 13/40 cases (33%) classified as luminal A were G3 with the PPH3 mitotic score and could benefit from chemotherapy. In conclusion, PPH3 improving MC gives a better categorization by halving the G2 group. In particular, applied to the surrogate subtype luminal A breast cancer it identified cases that could benefit from adjuvant cytotoxic chemotherapy.

The most frequent site of recurrence in breast cancer (BC) is the bone, particularly in patients with ‘luminal-like’ disease. Denosumab has been shown to prevent aromatase inhibitors (AIs) induced bone resorption in postmenopausal early BC patients and reduce skeletal-related events (SREs) in bone metastatic breast cancer (BMBC). A ‘real life’ analysis of 90 BMBC patients treated with denosumab was performed. Eighty-six patients (95.6%) had ‘luminal-like’ disease, 72 (80%) had bone metastases at the time of first recurrence of disease. Among 50 patients with metachronous ‘luminal-like’ disease, 40 (80%) had first recurrence to the bone. Among these patients median time to skeletal recurrence (TSkR) was shorter for patients who were previously exposed to AIs compared to those who were not (53.0 vs. 102.0 months, respectively; P=0.0300) and longer for patients previously treated with tamoxifen compared to those who were not (102.0 vs. 59.0 months, respectively; P=0.0466). Both of them were not confirmed at multivariate analysis. In the overall population, 17 first SREs were observed (16 radiation therapy) and median time to first SRE was not reached. A statistically significant difference in the incidence of SREs was detected only between patients with exclusively osteolytic bone metastases vs. those without (P=0.013). The presence of exclusively-osteolytic bone metastases was the only factor significantly associated with a shorter time to first SRE (P=0.011). The only G3 toxicity reported was hypocalcemia in one patient. No osteonecrosis of the jaw events (ONJ) occurred. This study demonstrated that a pro-active attitude enables the treatment of the majority of patients with denosumab without significant class-related toxicities. The majority of SREs were from radiation therapy, so pain still remains the clinical hallmark of bone metastases, particularly for osteolytic ones. The suggestion that estrogen deprivation with AIs can favor a ‘bone-related’ risk conditions for developing bone metastases must be considered with caution and surely needs further validations.

Candida albicans is implicated in intestinal diseases. Identifying host signatures that discriminate between the pathogenic vs commensal nature of this human commensal is clinically relevant. In the present study we have identified IL-9 and mast cells (MC) as key players of Candida commensalism and pathogenicity. By inducing TGF-β in stromal MC, IL-9 pivotally contributed to mucosal immune tolerance via the indoleamine 2,3-dioxygenase enzyme. However, Candida-driven IL-9 and mucosal MC also contributed to barrier function loss, dissemination and inflammation in experimental leaky gut models and were upregulated in patients with celiac disease. Inflammatory dysbiosis occurred in IL-9 and MC deficiency, a finding indicating that the activity of IL-9 and MC may go beyond host immunity to include regulation of the microbiota. Thus, the output of the IL9/MC axis is highly contextual during Candida colonization and reveals how host immunity and the microbiota finely tune Candida behavior in the gut.

Anaplastic lymphoma kinase (ALK) rearrangements represent the molecular driver of a subset of non-small cell lung cancers (NSCLCs). Despite the initial response, virtually all ALK-positive patients develop an acquired resistance to the ALK inhibitor crizotinib, usually within 12 months. Several next-generation ALK inhibitors have been developed in order to overcome crizotinib limitation, providing an unprecedented survival for this subset of patients. The aim of this review to summarize the current knowledge on ALK tyrosine kinase inhibitors (TKIs) in the treatment of advanced ALK-positive NSCLC, focusing on the role of novel ALK inhibitors in this setting. In addition, we will discuss their role in the pharmacological management of ALK-positive brain metastasis. Next-generation ALK inhibitors showed an impressive clinical activity in ALK-positive NSCLC, also against the sanctuary site of CNS. Sequential therapy with ALK TKIs appears to be effective in patients who fail a first ALK TKI and translates in clinically meaningful benefit. However, these agents display different activity profiles against crizotinib resistance mutation; therefore re-genotyping the disease at progression in order to administer the right TKI to the right patient is going to be necessary to correctly tailor the treatment. To avoid repeated invasive procedure, noninvasive methods to detect and monitor ALK rearrangement are under clinical investigation.

Distinguishing between alveolar rhabdomyosarcoma (ARMS) and embryonal rhabdomyosarcoma (ERMS) is crucial because treatment and prognosis are different. We describe a case of paratesticular rhabdomyosarcoma (RMS), which was classified as mixed ERMS/ARMS. Fluorescence in situ hybridization (FISH) detected losses of 3'PAX3 and 5'FOXO1, suggesting they had undergone an unbalanced rearrangement that probably produced the PAX3-FOXO1 fusion. Double-color FISH and reverse transcription-polymerase chain reaction (RT-PCR) revealed PAX3-FOXO1, which is characteristic of high-risk RMS. This finding highlights the importance of supplementing histology with genetics so that atypical RMS is appropriately classified and patients are correctly stratified and treated.

Amplification of genes of the MYC family (MYC, MYCN, and MYCL1) has been described in a variety of cancer cell lines and tumor specimens including lung cancer.1–4 The MYC gene localized at 8q24.1 is a well-characterized oncogene involved in cell growth, differentiation, metabolism, and apoptosis.5 The prognostic role of MYC amplification has been explored both in small cell lung cancer (SCLC) and non–small cell lung cancer (NSCLC). Lockwood et al6 have investigated 104 cancer cell lines from different tumor tissue origins by array comparative genomic hybridization to identify amplified chromosomal segments. Among 53 lung cell lines, of which 36 were originated from NSCLC, 16 from SCLC, and 1 from Mesothelioma, MYC was the most frequently amplified gene. Some studies on resected NSCLC reported that MYC was associated with tumor progression,7,8 a worse prognosis, and its overexpression was related to metastasis of lung cancer.9 Kubokura et al10 showed that MYC amplification correlated with lymph node metastasis, suggesting a possible negative effect on survival. Iwakawa et al11 showed that MYC amplification was associated with poor prognosis in patients both with small-sized (≤2 cm in greatest dimension) and early-stage I lung adenocarcinoma (ADC). Furthermore, MYC was expressed in large numbers of NSCLCs12 and was amplified or overexpressed in SCC and ADC of the lung.13–15 The MYC increased gene copy number (GCN) leads to overexpression of the MYC protein through Max heterodimer transcription factors that alter gene expression in large part by recruiting histone-modifying enzymes.15 The gain of sequences on the long arm of chromosome 3 (3q) is also a frequent event in many human malignant diseases, including lung cancer.2 Human telomerase gene (TERC), localized on the chromosome 3q26, encodes the RNA component of human telomerase, a ribonucleoprotein enzyme, which acts as a template for the addition of telomeric repeat sequences 5′-TTAGGG-3′16 required for the stability and complete replication of chromosome ends, and it has been associated with cell immortality and the development of cancers. Recently, Fan et al17 reported significantly higher percentages of cells with amplification of TERC in NSCLC than in SCLC, and the phenomenon occurred more frequently in SCC than in ADC.18 Using FISH methodology, Pelosi et al19 studied the 3q26 amplification in preneoplastic/preinvasive squamous cell lesions of the bronchial mucosa and in 2 subsets of lung SCC, the early hilar (EHSCC), and the parenchyma-infiltrating SCC (PISCC). The authors concluded that 3q26 amplification was likely a late event in the development of SCC of the lung and it is more prevalent in EHSCC than in PISCC, suggesting different pathogenesis for these tumor subtypes. Foster et al20 showed that the 3q26 amplification was a common feature of pulmonary SCC, confirming its key role in the transition from high-grade preinvasive neoplasia to invasive carcinoma, as also documented in uterine cervix21 and head-and-neck SCC.22 Yan et al23 reported that 3q and 8q amplifications were significantly higher in smoker than those in nonsmoker SCC patients and was associated with tumorigenesis and/or progression of the disease. In summary, only a few studies are available on MYC and TERC GCN in NSCLC, and no consensus criteria exist on how to assess the status of these genes as prognostic value. This study aimed to evaluate the MYC and TERC gene copy status in NSCLC by FISH using 2 criteria for interpretation, namely, the University of Colorado Cancer Center (UCCC) scoring system proposed for EGFR in lung cancer24 and the receiver operating characteristics (ROC) scoring system. We also evaluated the correlation of these genomic imbalances with clinicopathologic parameters and outcome in resected NSCLC patients.

Recent advances in the understanding of non-small cell lung cancer (NSCLC) biology have revealed a number of 'targetable' genetic alterations that underlie cancer growth and survival in specific patients subgroups. The anaplastic lymphoma kinase (ALK) gene rearrangement identifies a population of NSCLCs in whom dysregulation of ALK-tyrosine kinase (-TK) leads to uncontrolled proliferation of cancer cells, thus providing the basis for the therapeutic use of ALK-TK inhibitors (-TKIs) in ALK-rearranged (-positive) disease. Crizotinib was the first ALK-TKI to undergo clinical development in ALK-positive advanced NSCLC, in which it has been shown to greatly outperform the best available chemotherapy regimen in either second- or first-line setting. More recently, the novel second-generation ALK-TKI ceritinib has been shown to be highly active in either crizotinib-pretreated or -naïve population. Nevertheless, as mechanisms of resistance to crizotinib and ALK-TKIs in general are being progressively elucidated, the treatment landscape of ALK-positive NSCLC is expected to evolve rapidly. In the present review we will briefly discuss the current knowledge of ALK-positive advanced non-small cell lung cancer. Also, we will touch upon new developments on drugs/combination regimens aimed at inhibiting the ALK-TK, in an attempt to delineate how treatment of ALK-positive disease may change in the next future.

Background: Tumor-positive sentinel lymph node (SLN) biopsy results in a risk of non sentinel node metastases in micro-and macro-metastases ranging from 20 to 50%, respectively. Therefore, most patients underwent unnecessary axillary lymph node dissections. We have previously developed a mathematical model for predicting patient-specific risk of non sentinel node (NSN) metastases based on 2460 patients. The study reports the results of the validation phase where a total of 1945 patients were enrolled, aimed at identifying a tool that gives the possibility to the surgeon to choose intraoperatively whether to perform or not axillary lymph node dissection (ALND). Methods: The following parameters were recorded: Clinical: hospital, age, medical record number; Bio pathological: Tumor (T) size stratified in quartiles, grading (G), histologic type, lymphatic/vascular invasion (LVI), ER-PR status, Ki 67, molecular classification (Luminal A, Luminal B, HER-2 Like, Triple negative); Sentinel and non-sentinel node related: Number of NSNs removed, number of positive NSNs, cytokeratin 19 (CK19) mRNA copy number of positive sentinel nodes stratified in quartiles. A total of 1945 patients were included in the database. All patient data were provided by the authors of this paper. Results: The discrimination of the model quantified with the area under the receiver operating characteristics (ROC) curve (AUC), was 0.65 and 0.71 in the validation and retrospective phase, respectively. The calibration determines the distance between predicted outcome and actual outcome. The mean difference between predicted/observed was 2.3 and 6.3% in the retrospective and in the validation phase, respectively. The two values are quite similar and as a result we can conclude that the nomogram effectiveness was validated. Moreover, the ROC curve identified in the risk category of 31% of positive NSNs, the best compromise between false negative and positive rates i.e. when ALND is unnecessary (< 31%) or recommended (> 31%). Conclusions: The results of the study confirm that OSNA nomogram may help surgeons make an intraoperative decision on whether to perform ALND or not in case of positive sentinel nodes, and the patient to accept this decision based on a reliable estimation on the true percentage of NSN involvement. The use of this nomogram achieves two main gools: 1) the choice of the right treatment during the operation, 2) to avoid for the patient a second surgery procedure.

A large number of cancer predisposing BRCA1/BRCA2 mutations have been reported, with a wide variety among populations. In some restricted groups, specific germline mutations in these tumor suppressor genes have been found with high predominance, due to a founder effect. We focused our review on the Italian founder mutations. The first Italian BRCA1 founder mutation, 5083del19, was found in Calabria: the presence of common allele in all carriers of this mutation (also in families with Calabrian origin living in other parts of Italy) confirmed its founder effect. The same BRCA1 mutation was identified in the Sicilian population, but only the haplotype analysis can reveal the common ancestor of these groups. Another BRCA1 founder mutation, 4843delC, was found in Sicily. Four distinct BRCA1 mutations are attributable to families original from Tuscany: 3348delAG, 3285delA, 1499insA and 5183delTGT; the latter has been shown to be a founder mutation from North-Eastern Italy. The first BRCA2 mutation was identified in Sardinia, 8765delAG, a mutation already described as a founder mutation in Jewish-Yemenite families and also in French-Canadian population but with independent origins of carriers in these three populations. BRCA2 3951del3 and BRCA1 917delTT have been described as founder mutations in Middle Sardinia and in South and Middle Sardinia, respectively. Studies regarding prevalence and penetrance of founder mutations can allow to quantify the degree of homogeneity within a population and can surely help the geneticist and oncologist to simplify their choices in the genetic testing on high-risk families, on the basis of their ethnical origin.

Recent literature highlights the importance of pro-inflammatory cytokines in the biology of breast cancer stem cells, unravelling differences with respect to their normal counterparts. Expansion of mammospheres is a valuable tool for the in vitro study of normal and cancer mammary gland stem cells. Here, we expanded mammospheres from human breast cancer and normal mammary gland tissues, as well from tumorigenic (MCF7) and non tumorigenic (MCF10) breast cell lines. We observed that agonists for the retinoid X receptor (6-OH-hydroxidrofenantrene), retinoic acid receptor (all-trans retinoic acid) and peroxisome proliferator-activated receptor (PPAR)-γ (pioglitazone), reduce the survival of mammospheres generated from breast cancer tissues and MCF7 cells, but not from normal mammary gland or MCF10 cells. This phenomenon is paralleled by the hampering of pro-inflammatory Nuclear Factor-κB/Interleukin-6 axis that is hyperactive in breast cancer-derived mammospheres. The hindrance of such pathway associates with the down-regulation of mammospheres regulatory genes (SLUG, Notch3, Jagged1) and with the up-regulation of the differentiation markers estrogen receptor-α and keratin18. At variance, the PPARα agonist Wy14643 promotes mammospheres formation, up-regulating Nuclear Factor-κB/Interleukin-6 axis and mammosphere regulatory genes. These data reveal that nuclear receptors agonists (6-OH-hydroxidrofenantrene, all-trans retinoic acid, pioglitazone) reduce the inflammation dependent survival of breast cancer stem cells and that PPARα agonist Wy14643 exerts opposite effects on this phenotype.

1 alpha,25-dihydroxyvitamin D

Dose-dense chemotherapy is one of the treatments of choice for neoadjuvant therapy in breast cancer (BC). Activating mutations inWe conducted a phase I/II study of neoadjuvant, sequential, dose-dense anthracycline/taxane chemotherapy, plus trastuzumab in HER2-positive patients and investigated the correlation of pre-treatmentwe established a dose-dense docetaxel recommended dose of 60 mg/mThis study proposes an effective and safe neoadjuvant dose-dense anthracycline/taxane schedule and suggests that

Primary myelofibrosis (PMF) is an uncommon form of myeloproliferative neoplasm (MPN) characterized by a proliferation of predominantly megakaryocytes and granulocytes in the bone marrow that, in fully-developed disease, is associated with reactive deposition of fibrous connective tissue, extramedullary hematopoiesis (EMH), and splenomegaly. Kidney involvement is rare and clinically presents with proteinuria, nephrotic syndrome, and renal insufficiency. Renal damage can be due to EMH and glomerulopathy. Renal EMH presents three patterns: infiltration of the interstitium with possible renal failure caused by functional damage of parenchyma and vessels, infiltration of capsule and pericapsular adipose tissue, and sclerosing mass-like lesions that can cause hydronephrosis and hydroureter with obstructive uropathy and renal failure. Glomerulopathy associated with PMF is rarely described, ranging from 1 month to 18 years from diagnosis of the neoplasm to renal biopsy. It is characterized by expansion and hypercellularity mesangial, segmental sclerosis, features of chronic thrombotic microangiopathy (TMA), and intracapillary hematopoietic cells infiltrating in absence of immune-mediated glomerulonephritis. We present a nephrotic syndrome in PMF-related glomerulopathy, associated with EMH, without renal failure, in a patient under treatment for 2 years with JAK2 inhibitor ruxolitinib. Despite treatment, the patient died 7 months after renal biopsy. Nephrologists still know very little about this topic and there is no homogeneous data about incidence, pathogenesis, and optimal treatment of this poor prognostic PMF-associated nephrotic syndrome. We focus on data in the literature in the hope of stimulating hematologists, nephrologists, pathologists to future studies about the natural history of renal involvement, useful for optimal management of this rare pathology.

G2 ductal infiltrating carcinomas are a heterogeneous group of tumours with ambiguous clinical significance. This is because G2 carcinomas are almost always the largest category and poorly reproducible. Mitotic count (MC) is one of the causes of poor histological grading reproducibility. The phosphoistone H3 (PPH3) antibody improves identification of mitotic figures. The aim of our study is to demonstrate whether using a new histological grading system based on PPH3 immunostaining to assess MC can re-stratify G2 category. We selected 100 cases of G2 invasive carcinoma. The mitotic score was accurately re-evaluated performing MC on PPH3 immunostained sections. 21/100 G2 cases (21%) showed the same mitotic score both with hematoxilin and eosin (HE) and PPH3 while 79 cases (79%) with PPH3 shifted to a higher mitotic score. After re-grading the 100 G2 cases based on the assessment of mitotic score with PPH3 only 53 cases (53%) were confirmed as G2, while 47 cases (47%) had shifted to G3. Finally we reclassified early tumours in the surrogate molecular subtype according to the 2013 St. Gallen Conference criteria and found that 13/40 cases (33%) classified as luminal A were G3 with the PPH3 mitotic score and could benefit from chemotherapy. In conclusion, PPH3 improving MC gives a better categorization by halving the G2 group. In particular, applied to the surrogate subtype luminal A breast cancer it identified cases that could benefit from adjuvant cytotoxic chemotherapy.

Objectives Aquaporins (AQPs) are a group of transmembrane water-selective channel proteins thought to play a role in the regulation of water permeability for plasma membranes. Indeed, high AQP levels have been suggested to promote the progression, invasion and metastasis of tumours. Specifically, AQP1 and AQP5 overexpression in lung adenocarcinoma (AC) have been suggested to be involved in molecular mechanisms in lung cancer. The aim of this retrospective cohort single-centre study was to assess both the levels of expression and therein the prognostic significance, regarding outcome of AQP1 and AQP5 in resected AC patients. Methods Patients with histological diagnoses of lung AC submitted to pulmonary resection were included in this cohort study. Tissue microarrays containing cores from 185 ACs were prepared. AQP1 and AQP5 expressions were assessed by immunohistochemistry. Results were scored as either low (Score 0-2) or high (Score 3-9). Clinical data, pathological tumour-node-metastasis staging and follow-up were recorded. Multivariate Cox survival analysis and Fisher's t-test were performed. Results AQP1 overexpression was detected in 85 (46%) patients, while AQP5 overexpression was observed in 45 (24%) patients. AQP1 did not result being significantly correlated with clinical and pathological parameters, while AQP5 resulted more expressed in AC with mucinous and papillary predominant patterns. Patients with AQP1 overexpression had shorter disease-free survival (P = 0.001) compared with patients without AQP1 overexpression. Multivariate analysis confirmed that AQP1 overexpression was significantly associated with shorter disease-free survival (P = 0.001). Conclusions Our results evidenced that AQP1 overexpression resulted in a shorter disease-free survival in lung AC patients. Being so, AQP1 overexpression might be an important prognostic marker in lung AC.

Acquired resistance to tyrosine kinase inhibitors (TKIs) represents the Achilles' heel of targeted treatment in lung cancer. Epidermal growth factor receptor (EGFR)-TKIs are considered the standard first-line treatment for patients with EGFR mutant non–small cell lung cancer; however, after a median of 9 to 12 months, virtually all patients develop acquired resistance, which is mediated by the development of an EGFR -T790M secondary mutation in approximately 60% of cases. Different mechanisms of acquired resistance have also been described with lower incidence, including mutations in other driver oncogenes or phenotypic transformation. Herein, we report the first case of a patient with EGFR -mutant lung adenocarcinoma with a long-lasting response to first-line erlotinib treatment who acquired resistance to treatment because of acquisition of both EGFR -T790M mutation and "high-grade" large cell neuroendocrine transformation. This case also shows how resistance to third-generation EGFR-TKI osimertinib can be mediated by the development of phenotypic neuroendocrine transformation, which in the present case occurred during first-line treatment with erlotinib. In addition, our report highlights the pivotal role of rebiopsy and of molecular profiling at the time of progression to guide clinicians to choose the right therapy for the right patient.

// Valentina Cocciolone 1, 2 , Katia Cannita 1 , Maria Letizia Calandrella 1 , Enrico Ricevuto 2, 3 , Paola Lanfiuti Baldi 1 , Tina Sidoni 1 , Azzurra Irelli 1 , Stefania Paradisi 1 , Laura Pizzorno 4 , Valter Resta 4 , Alberto Bafile 4 , Edoardo Alesse 2 , Alessandra Tessitore 2 and Corrado Ficorella 1, 2 1 Medical Oncology, S. Salvatore Hospital, University of L'Aquila, Via Vetoio, 67100 L’Aquila, Italy 2 Department of Biotechnological and Applied Clinical Sciences, University of L’Aquila, Via Vetoio, 67100 L’Aquila, Italy 3 Oncology Network ASL1 Abruzzo, UOSD Oncology Territorial Care, S. Salvatore Hospital, University of L'Aquila, Via Vetoio, 67100 L’Aquila, Italy 4 Breast Unit, S. Salvatore Hospital, L’Aquila, Via Vetoio, 67100 L’Aquila, Italy Correspondence to: Valentina Cocciolone, email: vale.cocciolone@tiscali.it Keywords: clinicopathological prognostic factors, early breast cancer, clinical practice, survival, 20-year follow-up Received: November 08, 2016 Accepted: June 02, 2017 Published: June 16, 2017 ABSTRACT Background: To quantify the effect of traditional prognostic factors [nodal status, estrogen-receptor (ER), progesterone-receptor (PR), human epidermal growth factor receptor 2 (HER2)] on long-term outcome of patients with early breast cancer (EBC), treated in clinical practice over a period of about twenty years. Results: 1198 consecutive patients were identified. Median DFS (disease-free survival): ER+/PR±/HER2−, 165 months (mo) if node-negative (N0) and 114mo if node-positive (N+) ( p < 0.001); triple-negative (TN), 109mo if N0 and 65mo if N+ (p 0.144); ER+/PR±/HER2+ in patients not-treated with adjuvant trastuzumab (T−), not reached if N0 and 114mo if N+ (p 0.297); ER+/PR±/HER2+ in patients treated with trastuzumab (T+), 95mo if N0 and 85mo if N+ (p 0.615); ER−/PR−/HER2+ T−, not reached if N0 and 26mo if N+ (p 0.279); ER−/PR−/HER2+ T+, not reached if N0 and 66mo if N+ (p 0.014). Median OS (overall survival): ER+/ PR±/HER2−, 166mo if N0 and 144mo if N+ (p 0.028); TN, 158mo if N0 and 96mo if N+ (p 0.384); ER+/PR±/HER2+ T−, not reached if N0 and 157mo if N+ (p 0.475), ER+/PR±/HER2+ T+, not reached if N0 and 106mo if N+ (p 0.436); ER−/PR−/HER2+ T−, not reached if N0 and 34mo if N+ (p 0.273); ER−/PR−/HER2+ T+, not reached neither if N0 nor if N+ (p 0.094). Materials and Methods: Disease-free survival (DFS) and overall survival (OS) were evaluated according to tumor characteristics, based on information retrospectively retrieved from patients’ medical records. Conclusions: Pathological tumor characteristics and nodal status still represent useful tools in treatment selection and follow-up decision making of EBC patients in clinical practice.

Association among gene polymorphisms and clinical-pathological features. (DOC 99Â kb)

CMF/FEC treatment-related toxicity graded according to the NCI- CTC v.2.0. (DOC 55Â kb)

Background: Bladder cancer is very diffuse especially in men aged about 65 years and the diagnosis is difficult and invasive. The identification of simple urine biomarkers that will enable diagnosis at an earlier stage of disease and accurately monitor for recurrence is a big challenge. The aim of this research is to find a simple urine test that may substitute or reduce the invasive and expensive cystoscopy. Methods: Voided urines collected in the morning before operation from 25 patients affected by papillary bladder carcinoma (Ta) were used after a short conservation at −80 ℃ and long conservation for more than 1 year. The lipids were extracted and analyzed using an Agilent Technologies. A statistical analysis of the results was performed with the specific software LipoStar. Results: The results show a clear difference in lipids between patients and controls that can be separated into two groups. In order to identify the lipid species, characteristic fragments of MS/MS spectra were used. On that basis, about 50% of 150 lipids selected were identified. This subset is mainly formed by triacylglycerols, alkyl diacylglycerols, cholesteryl ester and palmitoyl-sphingomyelin that are all present in high quantity in patients’ urine. These differences become less evident in samples saved for more than 1 year. Although the dataset is small, it appears that the urine lipids may be used as markers with good specificity and sensitivity. Conclusions: The relevance of lipids in tumorigenesis as well the possible alterations of chromatin lipids are discussed including also the possibility of miRNA influence. The development of a special kit for use in screening and for monitoring the appearance or reappearance of bladder tumors could prevent many deaths due to this disease and may reduce the current invasive and expensive diagnostic system.

Glyoxalase 2 (Glo2), a metabolic enzyme, is overexpressed in some human cancers which suggests this enzyme may play a role in human tumorigenesis. In prostate cancer (PCa), the role of Glo2 has been scarcely investigated and there are no studies addressing a causative involvement of this protein in this neoplasia. Here, we examined the immunohistochemical profile of Glo2 in human PCa and benign adjacent tissues and investigated Glo2 involvement in PCa development in human prostate cell lines. PCa and matched adjacent normal tissues were obtained from paraffin sections of primary PCa from 20 patients who had undergone radical prostatectomy. Histopathological diagnosis was confirmed for each sample. Glo2 expression analysis was performed by immunohistochemistry in prostate tissues, and by qRT-PCR and immunoblotting in prostate cell lines. The causative and mechanistic role of Glo2 in prostate tumorigenesis was demonstrated by Glo2 ectopic expression/silencing and employing specific activators/inhibitors. Our results showed that Glo2 was selectively expressed in PCa but not in the luminal compartment of the adjacent benign epithelium consistently in all the examined 20 cases. Glo2 expression in PCa was dependent on androgen receptor (AR) and was aimed at stimulating cell proliferation and eluding apoptosis through a mechanism involving the p53-p21 axis. Glo2 was intensely expressed in the basal cells of benign glands but was not involved in PCa genesis. Our results demonstrate for the first time that Glo2 drives prostate tumorigenesis and suggest that it may represent a novel adjuvant marker in the pathological diagnosis of early PCa.

Association among gene polymorphisms and risk of toxicity of any grade (grade1â 2â 3-4 vs 0). (DOC 83Â kb)

Aim To assess the sensitivity of biparametric magnetic resonance imaging (bpMRI) with non-endorectal coil in the detection and localization of index (dominant) and non-index lesions in patients suspected of having prostate cancer. Patients and methods We carried-out a retrospective analysis of multiparametric MRI (mpMRI) of 41 patients who underwent radical prostatectomy. Results of MRI for detection and localization of index and non-index lesions were correlated with those of histology. Results No statistically significant difference in size was seen between tumor lesion at histology and index lesion at MRI. In 41 patients, a total of 131 tumors were identified at histology, while bpMRI (T2-weighted and diffusion-weighted MRI) approach detected 181 lesions. bpMRI gave 27.6% false-positives and 3.3% false-negatives. Sensitivity in lesion detection by bpMRI increased with lesion size assuming high values for lesions ≥10 mm. For bpMRI and mpMRI, the sensitivity for detecting index lesions was the same and equal: 100% in the peripheral zone 97.6% and 94.7% in the entire prostate and transitional zone, respectively. Conclusion bpMRI can be used alternatively to mpMRI to detect and localize index prostate cancer.