Search

Supplementary Table 2 from β3 Integrin Subunit Mediates the Bone-Resorbing Function Exerted by Cultured Myeloma Plasma Cells

Supplementary Table 1 from β3 Integrin Subunit Mediates the Bone-Resorbing Function Exerted by Cultured Myeloma Plasma Cells

αvβ3 integrin was investigated in multiple myeloma in relation to the in vitro osteoclast-like activity of malignant plasma cells. Myeloma cells from patients with skeleton involvement overexpressed αvβ3 and produced erosion pits on bone substrates, whereas this effect was not observed by cells from patients with no evidence of bone disease. We therefore explored the αvβ3 transcriptional pathway in the bone-resorbing cells. Silencing of β3 chain abrogated the ability to produce erosion pits and extracellular signal-regulated kinase 1/2 phosphorylation resulting in the defective function of cFos and nuclear factor activator T cell 1, the terminal effectors of osteoclast activation. A similar defect occurred in constitutively β3-deficient cells from patients with no skeleton disease. Microarray gene analysis of β3+ myeloma cells showed that several osteoclast-related genes were up-regulated. Their functions include the activation of receptor pathways β3 and c-fms that regulate several osteoclast functions. These data emphasize the postulated role of myeloma cells in multiple myeloma bone disease and suggest that their osteoclast-like activity is regulated, at least in vitro, by the β3 subunit of the integrin. [Cancer Res 2009;69(16):6738–46]

Background:Obesity is associated with lower serum vitamin D (25(OH)D) levels through several mechanisms. The aim of the study was to examine the possibility of a negative association between fat mass and 25(OH)D levels in a cohort of otherwise healthy overweight and obese subjects, independently of age, sex, blood pressure levels and anthropometric and metabolic parameters.Materials and Methods:147 overweight and obese subjects (106 women and 41 men), aged between 18 and 69 years, were enrolled into the study. All of them did not show any clinically evident metabolic or chronic diseases (i.e. hypertension, diabetes mellitus, renal failure, etc.) and did not use any kind of drug. Serum fasting levels of 25(OH)D, insulin, glucose, uric acid and lipids (triglycerides, total, HDL and LDL cholesterol) were measured. The season in which the blood samples were collected was autumn. Insulin resistance was assessed by using the Homeostasis Model Assessment (HOMA-IR). Body composition parameters (Fat Mass [FM], Fat Free Mass [FFM], body cell mass [BCM], Total Body Water [TBW]) were measured by electrical Bioimpedance Analysis (BIA). Lastly, demographic, anthropometric and clinical parameters (age, Body Mass Index [BMI], Waist Circumference [WC], Systolic (SBP) and Diastolic (DBP) blood pressure) were also assessed.Results:25(OH)D levels were significantly and negatively correlated with BMI (P Conclusion:This study clearly shows that 25(OH)D circulating levels are progressively lower with the increase of fat mass, independently of sex, body fat distribution, blood pressure and insulin and metabolic parameters. These data strongly show that adipose tissue accumulation per se is absolutely the main factor responsible factor for lower 25(OH)D levels in obese subjects, possibly through sequestration of fat soluble 25(OH)D in fat mass.

A significant increase in platelet count may be a risk factor for atherosclerotic cardiovascular disease. This study investigates the association between platelet number and glucose metabolism, evaluated by glycated hemoglobin (HbA1c) levels, in a apparently healthy population represented by overweight and obese subjects with normal glucose and HbA1c levels.As many as 240 subjects, 177 women and 63 men, aged 18-70 years, were enrolled. Body mass index (BMI), waist circumference (WC), systolic and diastolic blood pressure levels, platelet count and fasting blood glucose, insulin, insulin resistance, HbA1c, uric acid, triglyceride, total cholesterol, high and low density lipoprotein cholesterol concentrations were evaluated. Concerning the univariate correlation analyses between platelets number and all other variables, platelet count was significantly (and positively) correlated only with HbA1c (P 0.05) and female sex (P 0.01). HbA1c (P 0.05), female sex (P 0.001), and diastolic blood pressure (P 0.01), positively, and age (P 0.05) and systolic blood pressure (P 0.05), negatively, were significantly and independently associated to platelet count in a final multiple regression analysis.This is the first study showing a strong positive and independent relationship between HbA1c and platelet number in non-diabetic overweight and obese subjects.

A significant change of platelet number may be a risk factor for atherosclerotic cardiovascular disease. The aim of this study was to investigate the association between platelet number and early signs of atherosclerosis, evaluated by carotid intima-media thickness (c-IMT), in a apparently healthy population mainly represented by obese subjects.As many as 961 subjects, 686 women and 275 men, aged between 18 and 74 years, were enrolled in the study. Of these, 54 individuals (5.6% of all subjects) were normal weight, 259 individuals (27.0% of all subjects) were overweight, and 648 individuals (67.4% of all subjects) were obese. Waist circumference (WC) and blood glucose, insulin, total cholesterol, high and low density lipoprotein cholesterol, triglycerides and platelet count were also detected in all subjects, who underwent carotid echo color doppler ultrasound to measure c-IMT. c-IMT was significantly and positively associated to age (r = 0.204, P 0.0001), fasting glucose (r = 0.073, P 0.0240), total cholesterol (r = 0.096, P = 0.0031), and systolic and diastolic blood pressure (r = 0.140, P 0.0001 and r = 0.119, P 0.0003 respectively); c-IMT was significantly and negatively correlated with platelet count (r = -0.165, P 0.0001). Only age (P 0.0001) and systolic blood pressure (P = 0.0393), positively, and platelet number (P 0.0001), negatively, were significantly and independently associated to c-IMT in a final multiple regression analysis.Lower platelet number represented an independent determinant of c-IMT in a population, mainly represented by obese patients. These results suggest that a decrease of platelet number may well be an early defensive mechanism in subjects developing the thickening of carotid artery.

Immune markers in the peripheral blood of melanoma patients provide useful information for clinical management although there is poor consensus on circulating cells which could putatively reflect the disease activity and play a prognostic role. Here, we investigated both dendritic cells (DCs) and T-regulatory cells (Tregs). The number of DC subsets as myeloid (m) and plasmacytoid was measured by flowcytometry in 113 melanoma patients in different clinical stages and correlated with the disease activity to evaluate the recurrence free survival (RFS) calculated as difference between baseline and post-surgical values in relation to the criteria for the melanoma staging, as primary tumor removal, sentinel lymph node biopsy and completion of lymph node dissection. Circulating mDC levels were significantly lower in metastatic melanoma than in other stages and inversely correlated to Treg values while both populations were similarly expressed in inactive disease at stage I-III. Furthermore, the levels of these cells after melanoma removal were apparently related to the disease activity since their persistent defect reflected high risk of recurrence and reduced the RFS. This work highlighted the role of immune cell measurement for the management of melanoma activity and the identification of patients at potential risk of recurrence based on the mDC ratio.

Background Bone is the most frequent site of metastases from breast cancer (BC), but no biomarkers are yet available to predict skeletal dissemination. Methods We attempted to identify a gene signature correlated with bone metastasis (BM) onset in circulating tumour cells (CTCs), isolated by a DEPArray-based protocol from 40 metastatic BC patients and grouped according to metastasis sites, namely "BM" (bone-only), "ES" (extra-skeletal) or BM + ES (bone + extra-skeletal). Results A 134-gene panel was first validated through targeted RNA sequencing (RNAseq) on sub-clones of the MDA-MB-231 BC cell line with variable organotropism, which successfully shaped their clustering. The panel was then applied to CTC groups and, in particular, the "BM" vs "ES" CTC comparison revealed 31 differentially expressed genes, including MAF, CAPG, GIPC1 and IL1B, playing key prognostic roles in BC. Conclusion Such evidence confirms that CTCs are suitable biological sources for organotropism investigation through targeted RNAseq and might deserve future applications in wide-scale prospective studies.

Skeletal colonization is often regarded as a rare event in patients with neuroendocrine tumors (NETs) although both national registries and retrospective series report an incidence of bone metastases as high as 20% in subjects with advanced disease. While the biological mechanisms leading to bone metastatic colonization in NETs have been poorly investigated so far, key steps of osteotropic mechanisms, including the epithelial-to-mesenchymal transition, preparation of the premetastatic niche, migration of circulating tumor cells towards the bone marrow as well as the resulting alterations of the skeletal metabolism, are likely to operate also during the development of NET bone metastases. The skeleton involvement by NETs has a detrimental impact on both quality of life and patients’ prognosis, leading to pain in the majority of symptomatic subjects. While it is currently unclear whether or not the earlier recognition of bone involvement by PET/CT imaging techniques employing 68Ga-DOTA-conjugated peptides might improve outcomes through the exploitation of timely treatments, the management of bone-colonizing NETs is today based only on clinical experience from other osteotropic tumors. Here, we summarize the fundamental molecular mechanisms driving bone colonization and revisit both established and novel treatments for patients with bone metastatic NETs.

The clinical diagnosis of cancer emerges from the recognition of specific signs and symptoms, is corroborated by laboratory and imaging tests, and is eventually confirmed by pathology examination. While a comprehensive panel of laboratory tests may provide useful information in the diagnostic work-up of cancer patients, tumor markers should not be routinely investigated for diagnostic purposes, but their use should be confined to surveillance. CT and MRI scans represent the main radiologic techniques used for both anatomic diagnosis and staging of cancer, while 18F-FDG PET/CT allows the evaluation of the metabolic activity of tumor lesions. Innovative radiotracers including 11C-choline and 68Ga-DOTA compounds have been recently developed and show high specificity for detecting prostate cancer and neuroendocrine tumors, respectively. Novel biomarkers such as circulating tumor cells or circulating tumor DNA are on the horizon, and their clinical implementation is steadily undergoing. For the foreseeable future, the integration of next-generation sequencing technologies and high-resolution imaging modalities carries the promise of achieving earlier cancer diagnosis in the precision medicine era, as well as more accurate tumor staging and personalized tumor follow-up.

DEAD-Box Helicase 4 (Ddx4)+ ovarian stem cells are able to differentiate into several cell types under appropriate stimuli. Ddx4 expression has been correlated with poor prognosis of serous ovarian cancer (OC), while the potential role of Ddx4+ cells in non-serous epithelial OC (NS-EOC) is almost unexplored. The aim of this study was to demonstrate the presence of Ddx4+ cells in NS-EOC and investigate the effect of follicle-stimulating hormone (FSH) on this population. Increased Ddx4 expression was demonstrated in samples from patients with advanced NS-EOC, compared to those with early-stage disease. Under FSH stimulation, OC-derived Ddx4+ cells differentiated into mesenchymal-like (ML) cells, able to deregulate genes involved in cell migration, invasiveness, stemness and chemoresistance in A2780 OC cells. This effect was primarily induced by ML-cells deriving from advanced NS-EOC, suggesting that a tumor-conditioned germ cell niche inhabits its microenvironment and is able to modulate, in a paracrine manner, tumor cell behavior through transcriptome modulation.

Background: Circulating tumor cells (CTCs) have recently emerged as a new dynamic soluble marker for several malignancies including cutaneous melanoma (CM) and are suitable for prognostic evaluations and treatment monitoring. However, to date many limitations still hamper the wide-scale application of CTCs in CM setting, including the lack of standardized methods as well as both low levels and heterogeneity of these cells. Methods: We developed a protocol for CTC detection in CM based on immune-magnetic sorting to deplete CD45-, CD31- or CD34-positive cells, followed by dielectrophoretic DEPArray separation according to cell morphology and immunophenotype. To this end, we explored the expression of melanoma stem cell antigens (CD271, ABCB5, and RANK) and the epithelial-to-mesenchymal transition markers (N-Cad, -CD44, and -MCAM/CD146) on CTCs from 17 stage IV CM patients, and investigated their BRAF mutational status by droplet digital PCR. Results: The number of CTCs isolated from CM patients ranged from 2 to 91 cells (38 ± 6.4) with respect to healthy donors ( p V600 mutational status and matched primary tumors. The characterization of the immune phenotype of isolated cells revealed high interindividual and intraindividual heterogeneity that was found to correlate with the clinical outcome. Conclusions: The dual-step protocol of immune-magnetic sorting and subsequent dielectrophoretic DEPArray separation, turned out to be a suitable method to isolate viable CTCs from stage IV melanoma patients and enabled quantitative and qualitative analyses on these cells, which may deserve prospective evaluation for potential use in the clinical practice.

Aims Ischaemic heart disease is classically associated with coronary artery disease. Recent evidences showed the correlation between coronary microvascular dysfunction and ischaemic heart disease, even independently of coronary artery disease. Ion channels represent the final effectors of blood flow regulation mechanisms and their genetic variants, in particular of Kir6.2 subunit of the ATP-sensitive potassium channel (KATP), are reported to be involved in ischaemic heart disease susceptibility. The aim of the present study is to evaluate the role of KATP channel and its genetic variants in patients with ischaemic heart disease and evaluate whether differences exist between coronary artery disease and coronary microvascular dysfunction. Methods A total of 603 consecutive patients with indication for coronary angiography due to suspected myocardial ischaemia were enrolled. Patients were divided into three groups: coronary artery disease (G1), coronary microvascular dysfunction (G2) and normal coronary arteries (G3). Analysis of four single nucleotide polymorphisms (rs5215, rs5216, rs5218 and rs5219) of the KCNJ11 gene encoding for Kir6.2 subunit of the KATP channel was performed. Results rs5215 A/A and G/A were significantly more represented in G1, while rs5215 G/G was significantly more represented in G3, rs5216 G/G and C/C were both more represented in G3, rs5218 C/C was more represented in G1 and rs5219 G/A was more represented in G1, while rs5219 G/G was significantly more represented in G2. At multivariate analysis, single nucleotide polymorphism rs5215_G/G seems to represent an ischaemic heart disease independent protective factor. Conclusions These results suggest the potential role of KATP genetic variants in ischaemic heart disease susceptibility, as an independent protective factor. They may lead to a future perspective for gene therapy against ischaemic heart disease.

Most commonly described as sporadic, pulmonary adenocarcinoma with enteric differentiation (PAED) is a rare variant of invasive lung cancer recently established and recognised by the World Health Organization. This tumour is highly heterogeneous and shares several morphological features with pulmonary and colorectal adenocarcinomas. Our objective is to summarise current research on PAED, focusing on its immunohistochemical and molecular features as potential tools for differential diagnosis from colorectal cancer, as well as prognosis definition and therapeutic choice. PAED exhibits an ‘entero-like’ pathological morphology in more than half cases, expressing at least one of the typical immunohistochemical markers of enteric differentiation, namely CDX2, CK20 or MUC2. For this reason, this malignancy appears often indistinguishable from a colorectal cancer metastasis, making the differential diagnosis laborious. Although standard diagnostic criteria have not been established yet, in the past few years, a number of approaches have been addressed, aimed at defining specific immunohistochemical and molecular signatures. Based on previously published literature, we have collected and analysed molecular and immunohistochemical data on this rare neoplasm, and have described the state of the art on diagnostic criteria as well as major clinical and therapeutic implications.The analysis of data from 295 patients from 58 published articles allowed us to identify the most represented immunohistochemical and molecular markers, as well as major differences between Asian PAEDs and those diagnosed in European/North American countries. The innovative molecular approaches, exploring driver mutations or new gene alterations, could help to identify rare prognostic factors and guide future tailored therapeutic approaches to this rare neoplasm.

Enumeration of circulating tumor cells (CTCs) may reflect the metastatic potential of breast cancer (BC). By using the DEPArray, we investigated CTCs with respect to their epithelial-to-mesenchymal transition phenotype and compared their genomic heterogeneity with tissue biopsies. Seventeen stage IV BC patients were enrolled. Pre-enriched CTC suspensions were stained with fluorescent-labeled antibodies to epithelial (E) and mesenchymal (M) markers. CTC samples were processed by DEPArray system and clustered in relation to their markers. DNA from CTCs, as well as from primary tumor samples, was sequenced by next generation sequencing to assess the mutational state of 50 major cancer-related genes. We identified four different CTC subsets that harbored different gene variants. The most heterogenous CTC subsets included the M+/E− phenotype, which, however, expressed only 7 repeatedly mutated genes, while in the M−/E+ subset multiple mutations affected only 2 out of 50 genes. When matching all gene variants among CTC subsets, a small number of mutations was shared by only 4 genes, namely ATM, FGFR3, PIK3CA, and TP53 that, however, were absent in primary tumors. Our results postulate that the detected mutations in all CTC subsets may be considered as genomic markers of metastatic dissemination to be investigated during early stages of BC.

The accumulation of visceral body fat, has been shown to be associated with higher risk of metabolic and cardiovascular disease. This study was addressed to examine whether para- and perirenal fat thickness and epicardial fat thickness were correlated with anthropometric- and cardiometabolic risk factors. A cohort of 102 uncomplicated overweight and obese patients was examined. BMI, waist circumference, blood pressure, fasting insulin, glucose, triglycerides, total cholesterol, HDL-cholesterol, LDL-cholesterol serum levels, and insulin resistance (assessed by HOMAIR) were measured. Para- and perirenal fat thickness (PUFT) and epicardial fat thickness (EUFT) were measured by ultrasounds. PUFT was positively correlated with BMI (p

In the “precision medicine” era, chemotherapy still remains the backbone for the treatment of many cancers, but no affordable predictors of response to the chemodrugs are available in clinical practice. Single nucleotide polymorphisms (SNPs) are gene sequence variations occurring in more than 1% of the full population, and account for approximately 80% of inter-individual genomic heterogeneity. A number of studies have investigated the predictive role of SNPs of genes enrolled in both pharmacodynamics and pharmacokinetics of chemotherapeutics, but the clinical implementation of related results has been modest so far. Among the examined germline polymorphic variants, several SNPs of dihydropyrimidine dehydrogenase (DPYD) and uridine diphosphate glucuronosyltransferases (UGT) have shown a robust role as predictors of toxicity following fluoropyrimidine- and/or irinotecan-based treatments respectively, and a few guidelines are mandatory in their detection before therapy initiation. Contrasting results, however, have been reported on the capability of variants of other genes as MTHFR, TYMS, ERCC1, XRCC1, GSTP1, CYP3A4/3A5 and ABCB1, in predicting either therapy efficacy or toxicity in patients undergoing treatment with pyrimidine antimetabolites, platinum derivatives, irinotecan and taxanes. While formal recommendations for routine testing of these SNPs cannot be drawn at this moment, therapeutic decisions may indeed benefit of germline genomic information, when available. Here, we summarize the clinical impact of germline genomic variants on the efficacy and toxicity of major chemodrugs, with the aim to facilitate the therapeutic expectance of clinicians in the odiern quicksand field of complex molecular biology concepts and controversial trial data interpretation.

Exosomes (Exo) are small vesicles produced by melanoma cells and the accessory cells of the tumor microenvironment. They emerge via both classical and direct pathways and actively participate in tumor colonisation of distant tissues. The proteins, nucleic acids, cytokines and growth factors engulfed by Exo are transferred to recipient cells, where they drive numerous functions required for the tumor escape from immune system control and tumor progression. By positively or negatively modulating immune cell properties, Exo provoke immune suppression and, in turn, defective dendritic cell (DC) functions. Together, these effects limit the cytotoxicity of T-cells and expand both T-regulatory and myeloid-derived suppressor populations. They also hinder perforin and granzyme production by natural killer cells. Finally, Exo also control the organotropism of melanoma cells. The distinct phenotypic properties of Exo can be exploited both for diagnostic purposes and in the early identification of melanoma patients likely to respond to immunotherapy. The potential therapeutic application of Exo derived from DCs has been demonstrated in vaccination trials, which showed an increase in anti-melanoma activity with respect to circulating tumor cells. However, additional studies are required before Exo can be effectively used in diagnostic and therapeutic applications in melanoma.

Uterine carcinosarcoma (UCS), also known as malignant mixed Müllerian tumor, is a rare gynecological malignancy characterized by poor prognosis. This "biphasic" neoplasm presents an admixture of epithelial and mesenchymal/sarcomatoid tumor cells which partially share their molecular signature and exhibit a typical epithelial-to-mesenchymal transition gene expression profile. Due to the rarity of this cancer, at present there is a scarcity of specific treatment guidelines. Surgical resection remains the best curative option for localized disease, whereas the addition of peri-operative radiotherapy, chemotherapy and chemoradiation has been shown to further improve disease outcomes. In the metastatic setting, palliative chemotherapy is currently the treatment of choice, although no consensus exists about the best regimen to be delivered. Besides standard treatment options for the advanced disease, mechanistic insights into UCS pathogenesis and identification of its histopathological and molecular features boosted the development of novel, and potentially more effective, therapeutic agents, that will be here discussed.

Immunotherapy is one of the most recent systemic treatments to emerge for use in oncology, and is based on the blocking of inhibitory immune checkpoints to potentiate the immune response to cancer. The anti-cytotoxic T lymphocyte-associated antigen-4 antibody ipilimumab and anti-programmed cell death protein 1 antibodies, including nivolumab and pembrolizumab, are currently available and widely used, and other immune-inhibiting antibodies are now under intensive investigation. These antibodies have shown efficacy in a growing number of tumor types, following initial observations of their notable effects in melanoma treatment. Despite the efficacy of these antibodies, their novel mechanisms of action are also associated with a new class of side effects called immune-related adverse events (IRAEs). These side effects do not share a common pathophysiology with other anticancer treatments and, therefore, they often require specific therapies. When detected early and correctly treated, IRAEs are reversible; however, they can become severe and life-threatening if underestimated or inappropriately treated. This review aims to revisit the pathogenesis of IRAEs, with attention to gastrointestinal manifestations, since these are common and potentially dangerous complications of immunotherapy and represent a major cause of treatment discontinuation. Recommendations and guidelines for the management of IRAEs are also presented, in order to provide a clear and applicable algorithm for use by clinicians.

Background In the era of precision medicine, the suitability of fluoropyrimidine therapies in clinical oncology can be checked by pharmacogenetic investigations of single patients, thus optimizing resources and indicating the appropriate drugs to personalize their chemotherapy. For example, the presence of dihydropyrimidine dehydrogenase gene ( DPYD) polymorphisms in cancer patients may lead to adverse effects when adopting fluoropyrimidine-based therapies. Methods We detected in a cancer patient a rare germline synonymous heterozygous variant of DPYD (c.1905C>T) in proximity to the exon 14 splice donor site. Because in silico analyses hypothesized potential deleterious effects of the splice site, we performed both quantitative and qualitative mRNA analyses to investigate the possible pathogenic nature of the variant. Results We did not detect any alterations in mRNA expression or in the cDNA sequence of DPYD gene transcript. Conclusions Our observations suggest that the c.1905C>T variant of DPYD does not have a pathogenic effect. Therefore, assessment of the clinical significance of rare sequence variants could emphasize the predictive value of DPYD gene alterations in identifying patients at potential risk for fluoropyrimidine-related toxicity.

The in vitro ability of polyphenols, extracted from red grape, to modulate peripheral blood mononuclear cell responses has been evaluated in 20 obese (Ob) people. With regard to cytokine release in response to phorbol myristate acetate (PMA), levels of interleukin-2 (IL-2), interferon-γ(IFN-γ), IL-4, IL-10, and IL-17 were higher in the Ob than in healthy (H) subjects.Vice versa, IL-21 concentrations were detected only in H people but they were undetectable in the Ob counterpart. In general terms, levels of IL-1β, IL-6, IL-8, and tumor necrosis factor-αwere higher in Ob people when compared to H controls. On the other hand, polyphenols did not modify IFN-γ, IL-4, and IL-17 levels. However, an increase in IL-2 was observed in H individuals, whereas its levels were decreased in the Ob counterpart. Polyphenols significantly increased IL-10 release from H donors, whereas a trend to increase was observed in Ob people. In addition, polyphenols were able to significantly increase levels of H IL-21, while this was not the case in Ob people. Since IL-21 is an inducer of Th17 cells, it is likely that polyphenols may suppress the sources of this cytokineviaproduction of IL-10. Accordingly, polyphenols decreased IL-1βand IL-6 release in comparison to H controls.

Background: Primary pulmonary enteric adenocarcinoma (PEAC) is a rare non-small cell lung cancer subtype sharing morphologic and immunohistochemical features with colorectal adenocarcinoma. Given the frequency of lung metastases in colorectal cancer, the differential diagnosis of PEAC according to routine morphological and immunohistochemical findings may be difficult. Genome sequence by next-generation sequencing has recently introduced new perspectives to better define the diagnosis and tumor sensitivity to treatments, while the rarity of this subtype of cancer still limits the current knowledge of its molecular features and provides no information to address patients to tailored therapies. Methods: We diagnosed a rare case of subcutaneous metastasis as a first symptom of a PEAC. Formalin-fixed paraffin-embedded samples of the primary tumor and subcutaneous metastases were examined by immunohistochemistry, and subsequently by targeted next-generation sequencing analysis. Results: Morphological and immunohistochemical findings suggested a rare case of metastatic pulmonary adenocarcinoma with enteric aspects. Next-generation sequencing analysis performed on both the primary tumor sample and the cutaneous lesion identified two pathogenic variants on CDKN2A and KRAS in both of them. However, the metastasis showed two additional pathogenic mutations located in SMAD4 and FLT3 genes. Conclusions: We describe for the first time an extensive molecular analysis on a rare case of PEAC with an unusual cutaneous metastasis. Our observation suggests that a specific pattern of mutations is harbored in this neoplasm, and that additional molecular studies may provide further information to identify prognostic and hopefully predictive genes of response to treatment.

Ovarian cancer is a heterogeneous tumor with highly variable clinical features. However, it is diagnosed based on rigid, traditional criteria, and the prognosis is defined according to the major histomorphological pattern and degree of differentiation of the tumor. Recently, the gene expression profile, mutational status, epigenetic features, and gene drivers of ovarian cancer have been explored in molecular analyses, which has led to the identification of novel disease subtypes based on molecular signatures. In the clinical setting, these signatures not only allow an accurate prognostic prediction but also contribute to targeted therapy that includes angiogenesis inhibitors and small molecules in combination, or not, with conventional chemotherapy. Molecular signatures of ovarian cancer have also been detected in several studies investigating the genomics of high-grade serous tumors. The pro-angiogenic and mesenchymal nature of these signals suggests that major antitumor activity can be achieved with bevacizumab, as the prototypic antiangiogenic drug, and other angiogenesis inhibitors. This chapter summarizes current knowledge of ovarian cancer from a genomic perspective, focusing on the molecular subtypes of these tumors, as revealed using extended high-throughput analyses, as well as the latest personalized treatment options.

With the completion of the Human Genome Project in June 2000 and in step with the development of next generation -omics, a new era of medicine has started. In particular, the application and expansion of the molecular profiling of tumors have ushered in the development of personalized treatments, collectively referred to as precision cancer medicine (PCM), a field that is steadily growing. An increasing number of targetable molecules are being produced by the pharmaceutical industry, and their efficacy is strictly related to their specificity for reliable and well-characterized oncogenic pathways. The US Food and Drug Administration has therefore recommended that the development of these agents should be accompanied by companion diagnostics. The genomic revolution is also changing the way clinical trials of targeted drugs for cancer patients are being planned and conducted. The most common master protocols include basket trials, umbrella trials, and platform trials. Due to their flexibility and characteristics across molecularly defined tumor subtypes, novel drugs are included or excluded depending on their efficacy. Two examples on the use of precision diagnosis and treatment are described. The first one concerns patients with advanced non-small cell lung carcinoma, characterized by actionable or “druggable” oncogenic alterations and the sometimes remarkable improvement in objective response rates, progression-free survival, and overall survival compared with platinum-based chemotherapy. The second successful example of PCM is malignant melanoma. In parallel with a better understanding of the molecular mechanisms driving melanoma progression and drug resistance, the standard of care for this tumor has shifted from poorly effective chemotherapy to more optimistic targeted approaches. However, despite these encouraging results, a wider clinical application of PCM is still hampered by several drawbacks that will require major efforts before they can be overcome. It is hoped that, through the continuous and progressive refinement of its applications, PCM will eventually result in significant progress in the prevention, diagnosis, and treatment of tumors.

Cancer stem cells (CSCs) in patients with multiple myeloma (MM) are a subpopulation of tumor cells characterized by self-renewal capacity, clonogenic growth, and remarkable drug resistance and are thus considered the principal cause of tumor relapse. Although the exact phenotype of these cells has not been established, several markers, including CD138−, aldehyde dehydrogenase+, and side population cells, have been used to identify them. MM CSCs utilize common self-renewal pathways, such as Wingless, Hedgehog, Notch, and PI3K/Akt/mTOR, as well as bone marrow microenvironment-dependent pathways that can be targeted with novel drugs. Combination therapies with (1) agents that would target MM CSCs plus (2) conventional anti-MM drugs that affect the bulk of tumor plasma cells are expected to provide a promising strategy for the possible transformation of MM into a chronic disease and eventually for the cure of MM.

Management of localized well-differentiated pancreatic neuroendocrine tumors (panNETs) is controversial and primarily dependent on tumor size. Upfront surgery is usually recommended for tumors larger than 2 cm in diameter since they frequently show metastatic potential, whereas smaller panNETs are generally characterized by an indolent clinical course, with a rate of relapse or metastasis below 15%. To explore whether increased tumor size is paralleled by genomic variations, we compared the rate and the mutational patterns of putative driver genes that are recurrently altered in these tumors by investigating differential cohorts of panNET surgical specimens smaller (n = 27) or larger than 2 cm (n = 29). We found that the cumulative number of mutations detected in panNETs >2 cm was significantly higher (p = 0.03) relative to smaller tumors, while mutations of DAXX were significantly more frequent in the cohort of larger tumors (p = 0.05). Moreover, mutations of DAXX were associated with features of malignancy including increased grade, nodal involvement and lymphovascular invasion, and independently predicted both relapse after surgery (p = 0.05) and reduced DFS in multivariable analysis (p = 0.02). Our data suggest that alterations of the DAXX/ATRX molecular machinery increase the malignant potential of panNETs, and that identification of mutations of DAXX/ATRX in small, nonfunctioning tumors can predict the malignant progression observed in a minority of them.

Bone is the most common metastatic site for solid malignancies, especially those of the breast, lung, and prostate. These tumors produce severe skeletal complications, referred to as skeletal-related events, although their ability to colonize the skeleton is heterogeneous and many patients will not have bone metastasis (BM) of their tumors. Thus, extensive studies are in progress to define the osteotropism of bone-homing malignancies, a tumor property that can be evaluated by the identification of osteogenic and osteolytic soluble factors in biological fluids. However, these biomarkers reflect only the biology of BM and are useful in monitoring BM-induced disease, whereas novel surrogates allowing the early identification of osteotropic tumors in patients with otherwise identical tumor histotypes are needed to predict tumor progression and guide appropriate follow-up. Application of the omic sciences, either to tumor tissues or to circulating tumor cells (CTCs), and in several instances to peripheral blood, has led to the detection of a number of proteins and soluble factors involved in the epithelial-to-mesenchymal transition (EMT). This process underlies cancer cell migration to bone and is mediated by cytokines, chemokines, exosomes, miRNA, and components of bone morphogenetic proteins, among others. Preclinical studies of the EMT have yielded favorable preliminary results with potential translation to clinical investigations. In addition to the proteomic approach to osteotropism, “liquid biopsy” provides the basis for a novel application, as the subsequent isolation of CTCs or circulating tumor DNA followed by gene amplification suggests a route for the early prediction of the bone-metastasizing fate of solid cancers. By using RNAseq methodologies in a next-generation sequencing platform, the osteotropic behavior of these tumors can be recognized before the clinical detection of BMs, and patients at risk of skeletal tumor spreading can be appropriately treated. In the current era of precision medicine, in addition to the direct advantages of the early discovery of patients with tumors at risk of BM, the detection of a genomic “osteotropism signature,” whether in liquid or solid biopsy, will reduce imaging-based diagnostic costs for patients whose cancers are not genomically osteotropic.

Lung cancer is the leading cause of death from cancer. Some 80% of all lung cancers are lung adenocarcinomas, squamous cell carcinomas, and large cell carcinomas, which are grouped as non-small cell lung cancers (NSCLCs). The recent application of oncogenomics to NSCLCs has led to a radical change in the treatment of patients with advanced disease. Starting from the recognition of recurrent mutations in the tumor, new molecularly targeted therapies have been developed that have improved the outcomes of therapy for tailored subsets of patients with genetically homogeneous tumors. Roughly 45% of the mutations in NSCLC are point mutations involving the KRAS, EGFR, and BRAF genes; minor mutations giving rise to NSCLC consist of gene fusions that act as driver mutations in about 8% of cancers and involve the ALK, ROS1, and RET genes. While standard platinum-based chemotherapy remains the gold standard of treatment for NSCLCs without any targetable mutation, targeted tyrosine kinase inhibitors have yielded positive therapeutic outcomes in many patients, including those whose cancers involve gene translocations. This chapter reviews the major gene translocations associated with NSLCs, the main principles of therapy, important mechanisms of resistance, and the options available to overcome them.

The management of neuroendocrine tumors (NETs) has been rapidly evolving since the advent of the so-called “precision medicine” era. The availability of high-throughput sequencing technologies has substantially improved our ability to stratify NET patients based on their genomic, epigenomic, transcriptomic, or metabolomic profile. Although different molecular subgroups of NETs have been identified, prospective evaluation of their ability to predict prognosis, tumor clinical aggressiveness, and/or response to treatment is still lacking, thus hampering wide clinical application of such “-omic” profiles. Analysis of circulating NET transcripts as well as detection of changes in circulating tumor cells have been reported to predict response to treatment. If clinically validated in well-designed trials, liquid biopsy methodologies may be increasingly used in the future for both follow-up and treatment personalization. Molecular predictors of response to both somatostatin analogs, biologic agents, and chemotherapy have been intensely investigated, but no reliable biomarkers have been identified so far. An improved understanding of the complexity of pathway interactions through system biology approaches is required to advance further the NET field.

Neuroendocrine tumors (NETs) include a heterogeneous group of malignancies arising in the diffuse neuroendocrine system and characterized by indolent growth. Complex interactions take place among the cellular components of the microenvironment of these tumors, and the recognition of the molecular mediators of their interplay and cross talk is crucial to discover novel therapeutic targets. NET cells overexpress a plethora of proangiogenic molecules including vascular endothelial growth factor, platelet-derived growth factor, fibroblast growth factor, semaphorins, and angiopoietins that promote both recruitment and proliferation of endothelial cell precursors, thus resulting among the most vascularized cancers with a microvessel density 10-fold higher than epithelial tumors. Also, NETs operate multifaceted interactions with stromal cells, both at local and distant sites, and whether their paracrine secretion of serotonin, connective tissue growth factor, and transforming growth factor β primarily drives the fibroblast activation to enhance the tumor proliferation, on the other side NET-derived profibrotic factors accelerate the extracellular matrix remodeling and contribute to heart valves and/or mesenteric fibrosis development, namely, major complications of functioning NETs. However, at present, little is known on the immune landscape of NETs, but accumulating evidence shows that tumor-infiltrating neutrophils, mast cells, and/or macrophages concur to promote the neoangiogenic switch of these tumors by either direct or indirect mechanisms. On the other hand, immune checkpoint molecules are heterogeneously expressed in NETs’ surrounding cells, and it is unclear whether or not tumor-infiltrating lymphocytes are antitumor armed within the microenvironment, given their low mutational load. Here, we review the current knowledge on both gastroenteropancreatic and pulmonary NETs’ microenvironment as well as both established and innovative treatments aimed at targeting the tumor-host interplay.