Search

Introduction Bone metastases are frequent in patients with cancer. Electrochemotherapy (ECT) is a minimally invasive treatment. Preclinical and clinical studies supported the use of ECT in patients with metastatic bone disease (MBD). The purposes of this multicentre study are to confirm the safety and efficacy of ECT, and to identify appropriate operating procedures in different MBD conditions. Materials and methods 102 patients were treated in 11 Centres and recorded in the REINBONE registry (a shared database protected by security passwords): clinical and radiological information, ECT session, adverse events, response, quality of life indicators and duration of follow-up were registered. Results 105 ECT sessions were performed (one ECT session in 99 patients, two ECT sessions in 3 patients). 24 patients (23.5%) received a programmed intramedullary nail after ECT, during the same surgical procedure. Mean follow-up was 5.9 ± 5.1 months (range 1.5–52). The response to treatment by RECIST criteria was 40.4% objective responses, 50.6% stable disease and 9% progressive disease. According to PERCIST criteria the response was: 31.4% OR; 51.7% SD, 16.9% PD with no significant differences between the 2 criteria. Diagnosis of breast cancer and ECOG values 0–1 were significantly associated to objective response. A significant decrease in pain intensity and significant better quality of life was observed after ECT session at follow-up. Conclusion The results are encouraging on pain and tumour local control. ECT proved to be an effective and safe treatment for MBD and it should be considered as an alternative treatment as well as in combination with radiation therapy.

The major histocompatibility complex (MHC) class I expression in cancer cells has a crucial impact on the outcome of T cell-mediated cancer immunotherapy. We now determined the HLA class I allelic variants and their expression in PD-L1-deficient and positive rare sarcoma tissues. Tumor tissues were HLA-I classified based on HLA-A and -B alleles, and for class II, the HLA-DR-B by Taqman genomic PCRs. The HLA-A24*:10-B73*:01 haplotype was the most common. A general down-regulation or deletion of HLA-B mRNA and HLA-A was observed, compared to HLA-DR-B. HLA-I was almost too low to be detectable by immunohistochemistry and 32% of grade III cases were positive to PD-L1. Functional cytotoxic assays co-culturing patient biopsies with autologous T cells were used to assess their ability to kill matched tumor cells. These results establish that deletion of HLA-I loci together with their down-regulation in individual patient restrict the autologous lymphocyte cytotoxic activity, even in the presence of the immune checkpoint blocking antibody, Nivolumab. Additionally, the proposed cytotoxic test suggests a strategy to assess the sensitivity of tumor cells to T cell-mediated attack at the level of the individual patient.

The tumor microenvironment modulates cancer growth. Extracellular vesicles (EVs) have been identified as key mediators of intercellular communication, but their role in tumor growth is largely unexplored. Here, we demonstrate that EVs from sarcoma patients promote neoangiogenesis via a purinergic X receptor 4 (P2XR4) -dependent mechanism in vitro and in vivo. Using a proteomic approach, we analyzed the protein content of plasma EVs and identified critical activated pathways in human umbilical vein endothelial cells (HUVECs) and human progenitor hematopoietic cells (CD34+). We then showed that vessel formation was due to rapid mitochondrial activation, intracellular Ca2+ mobilization, increased extracellular ATP, and trafficking of the lysosomal P2XR4 to the cell membrane, which is required for cell motility and formation of stable branching vascular networks. Cell membrane translocation of P2XR4 was induced by proteins and chemokines contained in EVs (e.g. Del-1 and SDF-1). Del-1 was found expressed in many EVs from sarcoma tumors and several tumor types. P2XR4 blockade reduced EVs-induced vessels in angioreactors, as well as intratumor vascularization in mouse xenografts. Together, these findings identify P2XR4 as a key mediator of EVs-induced tumor angiogenesis via a signaling mediated by mitochondria-lysosome-sensing response in endothelial cells, and indicate a novel target for therapeutic interventions.

Sarcomas are rare, ubiquitous and heterogeneous tumors usually treated with surgery, chemotherapy, target therapy, and radiotherapy. However, 25–50% of patients experience local relapses and/or distant metastases after chemotherapy with an overall survival about 12–18 months. Recently, immuno-therapy has revolutionized the cancer treatments with initial indications for non-small cell lung cancer (NSCLC) and melanoma (immune-checkpoint inhibitors).Here, we provide a narrative review on the topic as well as a critical description of the currently available trials on immunotherapy treatments in patients with sarcoma. Given the promising results obtained with anti-PD-1 monoclonal antibodies (pembrolizumab and nivolumab) and CAR-T cells, we strongly believe that these new immunotherapeutic approaches, along with an innovative characterization of tumor genetics, will provide an exciting opportunity to ameliorate the therapeutic management of sarcomas.

Chondrosarcomas (CHS) are malignant cartilaginous neoplasms with diverse morphological features, characterized by resistance to chemo- and radiation therapies. In this study, we investigated the role of tumor-associated macrophages (TAM)s in tumor tissues from CHS patients by immunohistochemistry. Three-dimensional organotypic co-cultures were set up in order to evaluate the contribution of primary human CHS cells in driving an M2-like phenotype in monocyte-derived primary macrophages, and the capability of macrophages to promote growth and/or invasiveness of CHS cells. Finally, with an in vivo model of primary CHS cells engrafted in nude mice, we tested the ability of a potent peptide inhibitor of cell migration (Ac-d-Tyr-d-Arg-Aib-d-Arg-NH2, denoted RI-3) to reduce recruitment and infiltration of monocytes into CHS neoplastic lesions. We found a significant correlation between alternatively activated M2 macrophages and intratumor microvessel density in both conventional and dedifferentiated CHS human tissues, suggesting a link between TAM abundance and vascularization in CHS. In 3D and non-contact cu-culture models, soluble factors produced by CHS induced a M2-like phenotype in macrophages that, in turn, increased motility, invasion and matrix spreading of CHS cells. Finally, we present evidence that RI-3 successfully prevent both recruitment and infiltration of monocytes into CHS tissues, in nude mice.

Simple Summary Myxoid liposarcoma (MLPS) is the second most common subtype of liposarcoma, occurs predominantly in the extremities, and tends either to recur locally and metastasize to unusual soft tissue locations. To date, the mechanisms of invasion and metastasis of MLPS remain unclear, and yet, there is a high need to identify new prognostic biomarkers to enable the development of novel targeted therapeutic strategies. This study firstly aimed to assess the role of immune cellular components that infiltrate MLPS tissues. Our data show that high grade, heavily vascularized MLPS tissues exhibit T lymphocyte-poor and M2-like macrophage-rich phenotypes, while low grade MLPS tissues are mainly infiltrated by T lymphocytes. In line with these findings, evidence is shown that a crosstalk occurring between MLPS cells and macrophages exists as MLPS cells drive an M2-like phenotype in monocytes which in turn, increase the invasive capability of MLPS cells. Abstract Myxoid liposarcoma (MLPS) is the second most common subtype of liposarcoma and has tendency to metastasize to soft tissues. To date, the mechanisms of invasion and metastasis of MLPS remain unclear, and new therapeutic strategies that improve patients’ outcomes are expected. In this study, we analyzed by immunohistochemistry the immune cellular components and microvessel density in tumor tissues from patients affected by MLPS. In order to evaluate the effects of primary human MLPS cells on macrophage polarization and, in turn, the ability of macrophages to influence invasiveness of MLPS cells, non-contact and 3D organotypic co-cultures were set up. High grade MLPS tissues were found heavily vascularized, exhibited a CD3, CD4, and CD8 positive T lymphocyte-poor phenotype and were massively infiltrated by CD163 positive M2-like macrophages. Conversely, low grade MLPS tissues were infiltrated by a discrete amount of CD3, CD4, and CD8 positive T lymphocytes and a scarce amount of CD163 positive macrophages. Kaplan–Meier analysis revealed a shorter Progression Free Survival in MLPS patients whose tumor tissues were highly vascularized and heavily infiltrated by CD163 positive macrophages, indicating a clear-cut link between M2-like macrophage abundance and poor prognosis in patients. Moreover, we documented that, in co-culture, soluble factors produced by primary human MLPS cells induce macrophage polarization toward an M2-like phenotype which, in turn, increases MLPS cell capability to spread into extracellular matrix and to cross endothelial monolayers. The identification of M2-like polarization factors secreted by MLPS cells may allow to develop novel targeted therapies counteracting MLPS progression.

// Flavio Fazioli 1, * , Gianluca Colella 2, * , Roberta Miceli 3 , Mariano Giuseppe Di Salvatore 1 , Michele Gallo 1 , Serena Boccella 4 , Annarosaria De Chiara 5 , Carlo Ruosi 2, * and Filomena de Nigris 6, * 1 Division of Musculoskeletal Oncology Surgery, National Cancer Institute G. Pascale, Naples, Italy 2 Department of Human Health, Federico II University of Naples, Naples, Italy 3 S.C. Cellular Biology and Biotherapy, National Cancer Institute G. Pascale, Naples, Italy 4 Department of Experimental Medicine, University of Campania “Luigi Vanvitelli”, Naples Italy 5 Division of Pathology, National Cancer Institute G. Pascale Foundation, Naples, Italy 6 Department of Biochemistry, Biophysics and General Pathology, University of Campania “Luigi Vanvitelli”, Naples Italy * These authors have contributed equally to this work Correspondence to: Filomena de Nigris, email: Filomena.denigris@unina2.it Keywords: recurrences, sarcoma, angiogenesis, stem cells, AKT Received: March 31, 2017 Accepted: May 29, 2017 Published: June 28, 2017 ABSTRACT Giant cell tumors of bone (GCTB) are rare sarcomas with a high rate of unpredictable local relapse. Studies suggest that surgical methods affect recurrence, supporting the idea that local disease develops from re-growth of residual cancer cells. To identify early prognostic markers of individual risk of recurrence, we evaluated the effect of post-surgery fluids from a cohort of GCTB patients on growth of primary and established sarcoma cell lines, and mice xenograph. Post-surgery fluids increased cell growth and enhanced expression of CD44 ++ , the principal receptor for the extracellular matrix component hyaluronan and the mesenchymal stem marker CD117 + . Cancer cells became highly invasive and tumorigenic, acquiring stemness properties, and activated AKT/mTOR pathway. Prolonged stimulation with post-surgery fluids down-regulated the mesenchymal gene TWIST1 and Vimentin protein, and transdifferentiated cells into tubule-like structures positive to the endothelial markers VE-Cadherin and CD31 + . In mice, post-surgery fluids gave rise to larger and more vascularized tumors than control, while in patients AKT/mTOR pathway activation was associated with recurrence by logistic regression (Kaplan-Meier; P

Selection of cancer patients for treatment with immune checkpoint inhibitors remains a challenge due to tumour heterogeneity and variable biomarker detection. PD-L1 expression in 24 surgical chordoma specimen was determined immunohistochemically with antibodies 28-8 and E1L3N. The ability of patient-derived organoids to detect treatment effects of nivolumab was explored by quantitative and qualitative immunofluorescence and FACS analysis. The more sensitive antibody, E1L3N (ROC = 0.896, p = 0.001), was associated with greater tumour diameters (p = 0.014) and detected both tumour cells and infiltrating lymphocytes in 54% of patients, but only 1–15% of their cells. Organoids generated from PD-L1-positive patients contained both tumour cells and PD-1/CD8-positive lymphocytes and responded to nivolumab treatment with marked dose-dependent diameter reductions of up to 50% and increased cell death in both PD-L1-positive and negative organoids. Patient-derived organoids may be valuable to predict individual responses to immunotherapy even in patients with low or no immunohistochemical PD-L1 expression.

We describe radiographic, contrast-enhanced MDCT and MRI findings with pathologic correlations of an unusual recurrence of tumoral calcinosis, also called Teutschlander disease. The disease was silent in the first decade of life, when it appeared with elbows recurring lesions, until the seventh decade of life, when a left hip active growth lesion developed. A review about tumoral calcinosis pathogenesis, clinical course and imaging differential diagnosis is reported. (www.actabiomedica.it).

Yin Yang 1 (YY1) belongs to the polycomb group (PcG) of proteins that modify chromatin epigeneti-cally during dynamic regulation of their target genes. The predominant feature of YY1 is the zinc finger, an ancient structural motif that mediates protein–protein interactions and is capable of interacting with both DNA and RNA. Evidence reveals that YY1 acts predominantly as an epigenetic modulator, influencing the activity and/or localization of epigenetic modifiers molecules such as DNA methylation transferases, histone deacetylases, or non-coding RNAs. Deregulation of the epigenome is observed frequently in a variety of cancer types and is often correlated directly with advanced metastatic stages and poor prognosis. In this review, we address the current understanding of YY1 as a recruiter of epi-modifier molecules in the mechanism of aberrant regulation of target genes as a part of the metastatic cascade.

Cancer treatment is rapidly evolving toward personalized medicine, which takes into account the individual molecular and genetic variability of tumors. Sophisticated new in vitro disease models, such as three-dimensional cell cultures, may provide a tool for genetic, epigenetic, biomedical, and pharmacological research, and help determine the most promising individual treatment. Sarcomas, malignant neoplasms originating from mesenchymal cells, may have a multitude of genomic aberrations that give rise to more than 70 different histopathological subtypes. Their low incidence and high level of histopathological heterogeneity have greatly limited progress in their treatment, and trials of clinical sarcoma are less frequent than trials of other carcinomas. The main advantage of 3D cultures from tumor cells or biopsy is that they provide patient-specific models of solid tumors, and they overcome some limitations of traditional 2D monolayer cultures by reflecting cell heterogeneity, native histologic architectures, and cell– extracellular matrix interactions. Recent advances promise that these models can help bridge the gap between preclinical and clinical research by providing a relevant in vitro model of human cancer useful for drug testing and studying metastatic and dormancy mechanisms. However, additional improvements of 3D models are expected in the future, specifically the inclusion of tumor vasculature and the immune system, to enhance their full ability to capture the biological features of native tumors in high-throughput screening. Here, we summarize recent advances and future perspectives of spheroid and organoid in vitro models of rare sarcomas that can be used to investigate individual molecular biology and predict clinical responses. We also highlight how spheroid and organoid culture models could facilitate the personalization of sarcoma treatment, provide specific clinical scenarios, and discuss the relative strengths and limitations of these models.

Sacral chordoma is a rare low-to-intermediate grade malignant tumour. The mainstay of treatment is still surgery with en bloc and wide resection margins, which can grant the best chances of a long-term control or cure of this disease. The first aim of this paper is to collect data about survival, time to local recurrence and metastasis among patients affected by sacral chordoma and primarily treated with surgery. The second aim is to analyze the influence of level resection, tumor volume and surgical margins on local recurrence. The study population was composed of 14 patients treated with sacral chordoma resection at the National Tumour Institute of Naples—Pascale (Italy) from January 2000 to June 2013. The median follow-up was 84 months (range 24–132 months). The follow-up was characterized by: standard radiographs, MRI, and a CT scan of the chest annually. Time to recurrence or metastasis was calculated from the date of resection to the date of diagnosis of first recurrence or metastasis. Out of all the patients, six died (42.86 %) during the follow-up; 6 (42.86 %) had local recurrence; 4 (28.57 %) had metastasis. At univariate analysis wide surgical margins (R0) were associated with increased survival up to a local recurrence (OR = 0.0286; 95 % CI = 0.0014–0.5739; P = 0.026); the level of resection (OR = 3.33; 95 % CI = 0.3619–30.7025; P = 0.592) and tumour volume (P = 1) did not show a statistically significant correlation. Based on our experience, we hope all patients to be treated by surgery, the only good standard treatment of this disease. The resection should result in margins as wide as possible. For these reasons, it is essential for this disease to be treated in highly specialized centres because only a complete surgery can offer a chance to care for these patients. Solid survival at long-term follow-up can be achieved by a surgical resection performed with wide margins.

Background New materials (NM) such as titanium plates, cryopreserved grafts, and acellular collagen matrices are being increasingly used for chest wall reconstruction as a result of improved incorporation while maintaining structural stability and reduced need for removal from infected areas. Direct comparisons between NM and conventional materials (CM) in terms of local morbidity and need for prosthesis removal are lacking. Methods Between January 2005 and July 2013, 109 procedures were performed to remove chest wall tumors in 86 patients. Of these, 32 underwent complex chest wall reconstructions owing to either recurrence, defect extension (greater than 3 ribs or >100 cm 2 ) or local conditions (ie, previous irradiation or infection). New materials and CM (ie, polytetrafluoroethylene and methyl methacrylate) were used in 17 (53%) and 15 (47%) patients, respectively. Of the 32 patients included in the high complexity group, 23 patients did not exhibit any postoperative complications (72%). However, 9 patients (28%) underwent both a first and a second reoperation after a median interval of 4 months from the first procedure (range, 7 days to 60 months). Vacuum-assisted closure (VAC) was instituted in all patients as a means to control sepsis and facilitate space obliteration with healthy tissue. Results In 7 patients the reason for reintervention was local wound complications. In 4 of 7 patients, the prosthesis had to be removed (3 CM and 1 NM, 4.6% of the whole series; 12.5% in the high complexity group, 5.9% for NM and 20% for CM). The median time to complete chest wall healing after VAC in patients with local sepsis was 14 months (range, 5 to 60 months). All patients are currently alive and well except for 1 who died 11 months after complete chest wall healing as a result of dissemination of metastatic chondrosarcoma. At univariate analysis, predictors of overall and grade 2 or less morbidity according to the Common Terminology Criteria for Adverse Events version 4.0 were first ( p = 0.038) and second ( p = 0.015) redo operations. Conversely, patients with a body mass index of less than 25 kg/m 2 ( p = 0.049) undergoing one ( p = 0.032) or two reconstructions ( p = 0.00047) with combined materials ( p = 0.00029) were more likely to experience local wound complications and require VAC. On multiple regression analysis, redo operations (first, p = 0.032; second, p = 0.00047) and the use of combined (synthetic and biologic) materials ( p = 0.0029) were confirmed to be related to an increased incidence of wound complications. Conclusions Multiple redo operations after complex chest wall reconstruction performed with a combination of NM and CM may be associated with an increased incidence of local wound complications. Nevertheless, in these cases, the use of NM and VAC yielded a low rate (5.8% versus 20% with CM) of prosthesis removal while achieving complete wound healing.

Cooperation between endothelial cells and bone in bone remodelling is well established. In contrast, bone microvasculature supporting the growth of primary tumors and metastasis is poorly understood. Several antiangiogenic agents have recently been undergoing trials, although an extensive body of clinical data and experimental research have proved that angiogenic pathways differ in each tumor type and stage. Here, for the first time, we characterize at the molecular and functional level tumor endothelial cells from human bone sarcomas at different stages of disease and with different histotypes. We selected a CD31+ subpopulation from biopsies that displayed the capability to grow as adherent cell lines without vascular endothelial growth factor (VEGF). Our findings show the existence in human primary bone sarcomas of highly proliferative endothelial cells expressing CD31, CD44, CD105, CD146 and CD90 markers. These cells are committed to develop capillary-like structures and colony formation units, and to produce nitric oxide. We believe that a better understanding of tumor vasculature could be a valid tool for the design of an efficacious antiangiogenic therapy as adjuvant treatment of sarcomas.

Background.In several prospective and retrospective studies, weekly paclitaxel showed promising activity in patients with angiosarcoma.Patients and Methods. Our study was originally designed as a prospective, phase II multicenter trial for patients younger than 75, with ECOG performance status 0–2, affected by locally advanced or metastatic angiosarcoma. Patients received paclitaxel 80 mg/m2intravenously, at days 1, 8, and 15 every 4 weeks, until disease progression or unacceptable toxicity. Primary endpoint was objective response.Results. Eight patients were enrolled but, due to very slow accrual, the trial was prematurely stopped and further 10 patients were retrospectively included in the analysis. Out of 17 evaluable patients, 6 patients obtained an objective response (5 partial, 1 complete), with an objective response rate of 35% (95% confidence interval 17%–59%). Of note, five responses were obtained in pretreated patients. In the paper, details of overall survival, progression-free survival, and tolerability are reported.Conclusions. In this small series of patients with locally advanced or metastatic angiosarcoma, weekly paclitaxel was confirmed to be well tolerated and active even in pretreated patients.

// Vincenzo Ingangi 1, 2 , Katia Bifulco 1 , Ali Munaim Yousif 3 , Concetta Ragone 1, 2 , Maria Letizia Motti 4 , Domenica Rea 5 , Michele Minopoli 1 , Giovanni Botti 1 , Giuseppe Scognamiglio 6 , Flavio Fazioli 7 , Michele Gallo 7 , Annarosaria De Chiara 6 , Claudio Arra 5 , Paolo Grieco 3 , Maria Vincenza Carriero 1 1 Neoplastic Progression Unit, Department of Experimental Oncology, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy 2 SUN Second University of Naples, Naples, Italy 3 Department of Pharmacy, University Federico II, Naples, Italy 4 University ‘Parthenope’, Naples, Italy 5 Animal Facility, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy 6 Pathology Unit, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy 7 Surgery Unit, IRCCS Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy Correspondence to: Maria Vincenza Carriero, email: mariolinacarriero@yahoo.it , m.carriero@istitutotumori.na.it Keywords: urokinase receptor, formyl peptide receptor type 1, osteosarcoma, chondrosarcoma, peptides Received: April 18, 2016 Accepted: May 20, 2016 Published: June 13, 2016 ABSTRACT The receptor for the urokinase-type plasminogen activator (uPAR) is a widely recognized master regulator of cell migration and uPAR 88–92 is the minimal sequence required to induce cell motility and angiogenesis by interacting with the formyl peptide receptor type 1 (FPR1). In this study, we present evidence that the cyclization of the uPAR 88–92 sequence generates a new potent inhibitor of migration, and extracellular matrix invasion of human osteosarcoma and chondrosarcoma cells expressing comparable levels of FPR1 on cell surface. In vitro , the cyclized peptide [SRSRY] prevents formation of capillary-like tubes by endothelial cells co-cultured with chondrosarcoma cells and trans-endothelial migration of osteosarcoma and chondrosarcoma cells. When chondrosarcoma cells were subcutaneously injected in nude mice, tumor size, intra-tumoral microvessel density and circulating tumor cells in blood samples collected before the sacrifice, were significantly reduced in animals treated daily with i.p-administration of 6 mg/Kg [SRSRY] as compared to animals treated with vehicle only. Our findings indicate that [SRSRY] prevents three key events occurring during the metastatic process of osteosarcoma and chondrosarcoma cells: the extracellular matrix invasion, the formation of a capillary network and the entry into bloodstream.

High levels of urokinase receptor (uPAR) in tissue and serum of patients with chondrosarcoma correlate with poor prognosis. First, we analyzed the uPAR levels in tissues and plasma of five patients affected by chondrosarcoma. Interestingly, very high levels of uPAR and its soluble forms (SuPAR) were found on tumor cell surfaces and plasma, respectively, of two patients with lung metastases. Therefore, to investigate the role of SuPAR in chondrosaromas, we generated a primary cell culture from a chondrosarcoma tissue overexpressing uPAR on cell surfaces. We found that chondrosarcoma-like primary culture cells release a large amount of SuPAR in the medium.In vitro, SuPAR elicits chondrosarcoma cell migration likely through its uPAR88-92sequence, since the DII88-183or DIIDIIR88-284uPAR domains retain motogen effect whereas DI1-87or DIII184-284domains, both lacking the uPAR88-92sequence, are ineffective. Chondrosarcoma cells cross matrigel in response to SuPAR, and their invasion capability is abrogated by RERF peptide which inhibits uPAR88-92signalling. These findings assign a role to uPAR in mobilizing chondrosarcoma cells and suggest that RERF peptide may be regarded as a prototype to generate new therapeutics for the chondrosarcoma treatment.

This report concerns one case of sacrococcygeal chordoma and one case of parachordoma (soft-tissue chordoma) that were primarily diagnosed on fine-needle cytology (FNC) samples. Both neoplasms consisted of medium-sized epithelioid cells with eccentric, nucleolated nuclei, displaying abundant cytoplasm filled with bubbly vacuoles with refractile borders (physaliphorous cells). The neoplastic cells were embedded in an abundant extracellular substance staining metachromatically with Diff–Quik™. Both neoplasms had a typical clinical and instrumental presentation (primary sacrococcygeal tumor in a 58-year-old man and primary soft tissue mass of the hand palm and wrist in a 31-year-old man). The cytopathological findings in both the neoplasms were so similar as to be almost indistinguishable, and this similarity included the immunocytochemical findings. The deep similarity between these neoplasms indicates that the only pathological differences in the described cases probably reside in their different anatomical location and, perhaps, in different cytogenetic changes. Diagn. Cytopathol. 2010. © 2010 Wiley-Liss, Inc.

Giant cell tumors are mostly benign but locally aggressive tumors. The excision of bone tumors can result in large defects, therefore bone reconstruction is still one the most demanding procedures in orthopedic surgery. Our study addresses the opportunity for improving surgical outcome by employing β-tricalcium phosphate (β-TCP) with platelet-rich plasma (PRP) at the surgical site.We included 16 patients with giant cell tumors. After adjuvant therapy, the cavity was reconstructed with β-TCP, bone graft material (ActifuserR Granules Baxter) and platelet gel application.Our explorative analysis suggests a positive effect of PRP on surgical outcome in patients with giant cell tumors treated with curettage.Use of platelet gel as an adjuvant significantly reduces the time required for bone healing following intralesional treatment of benign giant cell tumors, and achieves good functional results without promoting local recurrence.

BACKGROUND: We recently developed a clinical decision support tool, capable of estimating the likelihood of survival at 3 and 12 months following surgery for patients with operable skeletal metastases. After making it publicly available on www.PATHFx.org , we attempted to externally validate it using independent, international data. METHODS: We collected data from patients treated at 13 Italian orthopaedic oncology referral centers between 2010 and 2013, then applied to PATHFx, which generated a probability of survival at three and 12-months for each patient. We assessed accuracy using the area under the receiver-operating characteristic curve (AUC), clinical utility using Decision Curve Analysis (DCA), and compared the Italian patient data to the training set (United States) and first external validation set (Scandinavia). RESULTS: The Italian dataset contained 287 records with at least 12 months follow-up information. The AUCs for the three-month and 12-month estimates was 0.80 and 0.77, respectively. There were missing data, including the surgeon's estimate of survival that was missing in the majority of records. Physiologically, Italian patients were similar to patients in the training and first validation sets. However notable differences were observed in the proportion of those surviving three and 12-months, suggesting differences in referral patterns and perhaps indications for surgery. CONCLUSIONS: PATHFx was successfully validated in an Italian dataset containing missing data. This study demonstrates its broad applicability to European patients, even in centers with differing treatment philosophies from those previously studied.

Background: Poor bone regeneration is a devastating complication after large tumor resection. Different synthetic bone-grafting materials have been used to restore skeletal defects. Several studies evaluated the capability of mesenchymal stem cells to improve the efficacy of bone regenerative potential of biomaterials. However, there are no data, in human, about the combinatory use of silicate granules with autologous Bone Marrow Mononuclear cells (BMMCs) to refill bone cavity. Methods: BMMCs were prepared in accordance with the International Society for Cell Therapy guidelines. MTT assays and scanning electron microscopy evaluated the biocompatibility, and adhesion of BMMCs to the granular silicate bone substitute. In vivo study is based on twenty patients with malignant osteolytic lesions. The mean volume of the lesions, measured by pre-operative computed tomography (CT) was 18.5 cm2 (range 16-24 cm2). Sixteen patients were treated post-surgery with curettage and refilling with granular silicate alone, and four with a combination of granular silicate and BMMCs The follow-up was determined by clinical, Musculoskeletal Tumor Society functional state score system (MSTS), and radiograph examination, moreover, the callus type was classified according to the International System. Results: Scanning electron microscopy showed that 90% of BMMCs adhered to silicate granules in a few minutes and long pseudopodia contacting extra-cellular matrix. Patients treated with autologous BMMCs and granular silicate developed bone callus after two weeks. The follow-up of limbs functional state measured by MSTS was a mean of 82% compared to mean of 60% obtained with granules alone. At the end of the follow-up (minimum one year), all the patients were cancer-free with an excellent outcome. Conclusions: The encouraging results of our early study indicate that refilling at the osteotomy site with autologous BMMCs and granular silicate improves bone repair. A larger study cohort and longer follow-up times are required to identify additional predictors and indications.

Osteosarcoma is the most common primary malignant bone tumour in children and adolescents.1–3 Although osteosarcoma is primarily a disease of adolescents and young adults, a different type of osteosarcoma linked to Paget’s disease occurs in older adults. The outcome for patients with osteosarcomas is poor with a 2-year overall survival rate of 15–20% following surgical resection, chemotherapy and radiotherapy.2,4,5 The most frequent site for metastasis is the lung.6 However, metastasis can also occur in other bones and soft tissue.6–9 Death fromosteosarcoma is usually the result of progressive pulmonary metastasis with respiratory failure due to widespread disease.1,10,11 Increasing the understanding of the basic biology of osteosarcoma is a priority for tumour prognosis as well as for providing indications of possible tumour targets for selective therapy. The transcription factor Yin Yang 1 (YY1) is a multifunctional DNA binding protein and an ubiquitously expressed member of a polycomb group of proteins.12–14 The transcription factor YY1 is known to be present in some human cancer cell lines and its expression correlates to immune-mediated apoptosis.15–20 Here, we demonstrate for the first time that the transcription factor YY1 is selectively expressed in human osteosarcoma cells and biopsies. The RT-PCR analysis confirmed the data obtained with immunohistochemistry. 0959-8049/$

Doxorubicin and ifosfamide are the two most active drugs in the treatment of patients with advanced, soft tissue sarcoma (STS) of most histologic subtypes, aside from gastrointestinal stromal tumor (GIST). However, after failure of these drugs, alone or in combination, patients with advanced STS have few therapeutic options and the search for new active drugs is well worth pursuing. ET-743, a DNA minor groove binder, which blocks cell cycle progression in G2/M phase through a p53-independent apoptotic process, represents the most promising among novel compounds in STS, since recently completed phase II trials have consistently shown high survival, in spite of the relatively low incidence of major objective responses. The potential for combination with other active compounds further increases the appeal of ET-743. Imatinib mesylate is being tested also in STS other than GIST, which can overexpress one or more of the tyrosine kinases inhibited by imatinib; however, negative data have recently been presented. Clinical studies with a number of other compounds are ongoing or planned. However, investigators involved in the management of patients with advanced STS are to be increasingly aware of the emergence of new molecular targets and genetic profiles in different histologic subtypes, according to which treatment strategies should be adapted.

Palmoplantar keratoderma (PPK) refers to a genetically heterogeneous group of skin diseases, which may be inherited in autosomal dominant or recessive fashion. We observed a case of a 74-year-old man with Mal de Meleda, who developed malignant melanoma inside the hyperkeratotic palmar skin of the right hand. Many authors have reported a higher incidence of cancer in cases affected by palmoplantar hyperkeratosis both for hereditary association and particularly for mechanical damage of the affected areas. The association with melanoma has already been described, but not in a true Mal de Meleda type syndrome as in the case reported in this paper.

Cryopreserved materials have characteristics which make them suitable for use in chest wall reconstruction. This contribution depicts the use of a cryopreserved bone homograft as a neosternum in conjunction with other materials to preserve anatomo-functional integrity and protect inner viscera. The final outcome and the emerging evidence in the literature support further utilization of these materials in the future.

Primary sarcomas represent a serious oncologic challenge because, at times, their surgical removal creates large defects requiring covering with several autologous, homologous, or prosthetic materials used in 1 or 2 graft combinations. 1-4 Although the experience is more limited, relapsing sarcomas often call for even more extensive demolitions of the chest wall and difficult reconstructive.

Liposarcoma (LPS) is the most common soft tissue neoplasm in adults and is characterized by neoplastic adipocyte proliferation. Some subtypes of LPSs show aberrations involving the chromosome 12. The most frequent are t(12;16) (q13;p11) present in more than 90% of myxoid LPSs and 12q13-15 amplification in well-differentiated and dedifferentiated LPSs. In this region, there are important oncogenes such as CHOP (DDIT3), GLI, MDM2, CDK4, SAS, HMGA2, but also the HOXC locus, involved in development and tumor progression. In this study, we evaluated the expression of HOXC13, included in this chromosomal region, in a series of adipocytic tumors. We included 18 well-differentiated, 4 dedifferentiated, 11 myxoid and 6 pleomorphic LPSs as well as 13 lipomas in a tissue microarray. We evaluated the HOXC13 protein and gene expression by immunohistochemistry and quantitative PCR. Amplification/translocation of the 12q13-15 region was verified by FISH. Immunohistochemical HOXC13 overexpression was observed in all well-differentiated and dedifferentiated LPSs, all characterized by the chromosome 12q13-15 amplification, and confirmed by quantitative PCR analysis. In conclusion, our data show a deregulation of the HOXC13 marker in well-differentiated and dedifferentiated LPSs, possibly related to 12q13-15 chromosomal amplification.

The combination of titanium plates and acellular collagen matrix was used to restore anterior chest wall integrity in a 68-year-old woman more than 30 years after a Halsted mastectomy and radiation treatment. A vertical rectus abdominis muscle flap was used for myocutaneous coverage of the reconstructed chest wall. Partial necrosis of the flap caused prolonged exposure of the collagen matrix and the titanium plates, which were then covered with a free skin graft. Long-term results were satisfactory despite continued infection treated with vacuum-assisted closure and surgical debridement. Recently introduced materials for chest wall reconstruction may offer resilience to infection, tolerability, and stability, and their use may be contemplated when the potential for local morbidity is high.

Background The polycomb transcription factor Yin Yang 1 (YY1) overexpression can be causally implicated in experimental tumor growth and metastasization. To date, there is no clinical evidence of YY1 involvement in outcome of patients with osteosarcoma. Prognosis of osteosarcoma is still severe and only few patients survive beyond five years. We performed a prospective immunohistochemistry analysis to correlate YY1 immunostaining with metastatic development and survival in a selected homogeneous group of patients with osteosarcoma. Methods We studied 41 patients suffering from osteosarcoma (stage II-IVa). Multivariate analysis was performed using Cox proportional hazard regression to evaluate the correlation between YY1 expression and both metastasis development and mortality. Results YY1 protein is not usually present in normal bone; in contrast, a high number of patients (61%) showed a high score of YY1 positive cells (51-100%) and 39% had a low score (10-50% positive cells). No statistical difference was found in histology, anatomic sites, or response to chemotherapy between the two degrees of YY1 expression. Cox regression analysis demonstrated that the highest score of YY1 expression was predictive of both low metastasis-free survival (HR = 4.690, 95%CI = 1.079-20.396; p = 0.039) and poor overall survival (HR = 8.353, 95%CI = 1.863-37.451 p = 0.006) regardless of the effects of covariates such as age, gender, histology and chemonecrosis. Conclusion Overexpression of YY1 in primary site of osteosarcoma is associated with the occurrence of metastasis and poor clinical outcome.

none 8 no This study aimed to identify, isolate, and characterize cancer stem cells from human primary sarcomas. We performed cytometric analyses for stemness and differentiation antigens, including CD29, CD34, CD44, CD90, CD117, and CD133, on 21 human primary sarcomas on the day of surgery. From sarcoma biopsies, we obtained 2 chondrosarcoma-stabilized cell lines and 2 osteosarcoma stabilized cell lines, on which sphere formation, side population profile, stemness gene expression, and in vivo and in vitro assays were performed. All samples expressed the CD133, CD44, and CD29 markers. Therefore, we selected a CD133 subpopulation from stabilized cell lines that displayed the capacity to grow as sarcospheres able to initiate and sustain tumor growth in nonobese diabetic/severe combined (NOD/SCID) mice, to express stemness genes, including OCT3/4, Nanog, Sox2, and Nestin, and to differentiate into mesenchymal lineages, such as osteoblasts and adipocytes. Our findings show the existence of cancer stem cells in human primary bone sarcomas and highlight CD133 as a pivotal marker for identification of these cells. This may be of primary importance in the development of new therapeutic strategies and new prognostic procedures against these highly aggressive and metastatic tumors.—Tirino, V., Desiderio, V., Paino, F., De Rosa, A., Papaccio, F., Fazioli, F., Pirozzi, G., Papaccio, G. Human primary bone sarcomas contain CD133 cancer stem cells displaying high tumorigenicity in vivo. FASEB J. 25, 2022–2030 (2011). www.fasebj.org Tirino V; Desiderio V; Paino F; De Rosa A; Papaccio F; Fazioli F; Pirozzi G; Papaccio G. Tirino V; Desiderio V; Paino F; De Rosa A; Papaccio F; Fazioli F; Pirozzi G; Papaccio G.

Aims and Background The prognosis of each individual patient affected by sarcoma, including those with low histopathologic grading, cannot be reliably predicted at the time of surgery. We have developed an in vitro cell invasion assay on early primary cell cultures derived from surgically removed sarcomas. Methods Primary cell cultures were subjected to in vitro cell invasion assays by using Boyden chambers, filters coated with matrigel and fetal bovine serum as a source of chemoattractant. For each primary cell culture, the sarcoma cell invasion index was determined in comparison with the percentage of human fibrosarcoma HT1080 cell invasion extent. The cell invasion index of 7 different sarcomas was evaluated in respect to the outcome of the disease, after a follow-up ranging from 14 to 48 months. Results Data evidenced that a low cell invasion index (39.7% ± 8.9) was retained by tumor cells derived from patients with no progression of the disease and with a longer interval of disease-free survival (21 ± 0.8 months). However, an increase in cell invasion index (61% ± 5) was retained by tumor cells derived from patients with progression of the disease and with a shorter disease-free survival (9 ± 3 months). Overall, although only 7 cases were analyzed, a statistically significant correlation was found between disease-free survival and cell invasion index (P = 0.003). Conclusions Our data support the possibility that cell invasion assays performed in vitro on cells derived from human sarcomas may be predictive of a more aggressive form of the disease.

The occurrence of a primary intramuscular infestation of Echinococcus granulosus is extremely rare.A 70-year-old woman with primary skeletal muscle hydatidosis initially presented with a soft tissue mass. Clinical and radiologic examination revealed a huge cystic mass in the right quadriceps muscle without any visceral organ involvement. Since the differential diagnosis included a soft tissue tumor, fine needle aspiration cytology was performed, and a diagnosis of hydatid disease was made.This very rare case of primary intramuscular infestation of E granulosus was clinically misdiagnosed as a soft tissue tumor. Hydatid disease, albeit rare, should be considered in the differential diagnosis of a soft tissue mass.

Adult soft tissue sarcomas (STSs) are a rare group of highly heterogeneous neoplasms arising in different tissues. They are locally aggressive and can produce recurrence and distant metastasis. The most common metastatic sites are lung, lymph nodes, liver, bone and soft tissues. Staging for STSs has been based on some prognostic information: grade (low vs. intermediate/high grade), size (small vs. large tumors), depth of infiltration (superficial vs. deep neoplasms) and presence or not of distant metastasis. In the last 10 years, a plethora of new markers (proliferation markers and DNA alteration, P-gp, p53, TLS-CHOP, cyclins, survivin, TERT, PAX3-PAX7/FKHR, SYT-SSX1/2, VEGF, E-cadherin and beta-catenin, nm23, SKP-2, p27, CD40) has been studied with regard to their role in promoting progression (in a laboratory setting) and then determining prognosis and therapy (in a clinical setting). In the present survey, we focused on the role of new biological prognostic factors in STSs and also reported the quality of such studies with an ad hoc designed questionnaire.

Purpose: The purpose is to evaluate the expression of CD40, a membrane protein predominantly expressed on B cells, dendritic cells, and macrophages, in a series of adult soft tissue sarcomas and to test its possible prognostic value. Experimental Design: CD40 expression was studied by immunohistochemistry. Correlations with other baseline characteristics of patients and tumors were analyzed with χ2 test. The prognostic value was studied with univariable and multivariable analysis adjusted by age, sex, tumor size, grade, location, and distant metastases. Results: Eighty-two patients, between January 1994 and May 2001, were analyzed. Membrane or cytoplasmic staining for CD40 protein was absent in 30% of the tumors but present in 50% of cells in 12 (15%) tumors. There was no correlation between CD40 expression and age, sex, size, grade, and location of the primary tumor and distant metastases. With 61 patients (74.4%) progressed and 31 (37.8%) dead, CD40 expression was a significant prognostic factor for disease-free and overall survival at univariable and multivariable analysis. Patients with tumors expressing CD40 in >50% of cells had a dramatically unfavorable prognosis with median disease-free and overall survival of 7 and 17 months, respectively, and hazard ratios of relapse and death as compared with patients with CD40-negative tumors of 2.89 (95% confidence interval: 1.26–6.60) and 6.92 (95% confidence interval: 2.18–22.0), respectively. Conclusions: These data suggest that expression of CD40 protein in >50% of cells might indicate an unfavorable prognosis in adult soft tissue sarcomas.

A 38-year-old man was diagnosed with fibrous dysplasia of the anterolateral segment of the fifth rib by core biopsy. A decision was made to resect the rib by video-assisted thoracic surgery (VATS) taking care to preserve the muscle and overlying myodermal layers. Subsequent reconstruction was done using a straight titanium plate locked in place under thoracoscopic guidance to avoid friction of the plate on the skin and to verify that the transfixed screws would not injure the lung or the pericardium. The patient made an uneventful recovery and was dismissed on day 2 after surgery. Final diagnosis confirmed fibrous dysplasia.

Among the new materials introduced for chest wall reconstruction, the use of collagen matrix is gaining increasing favor for its biomechanical properties. We describe the reconstruction of the chest wall with Veritas (Synovis, St Paul, MN) collagen matrix of a posterior chest wall defect after costovertebrectomy for Ewing's sarcoma. En bloc resection was performed, including partial D7 through D9 vertebrectomy along with the posterolateral segments of corresponding ribs. The collagen matrix patch was sutured to the spine stabilizer and the surrounding rib segments and was covered by previously raised latissimus dorsi and trapezius muscle flaps. Excellent stabilization was obtained.

From the Department of Thoracic Surgery and Oncology, Division of Thoracic Surgery, Services of Orthopaedic and Plastic Surgery, National Cancer Institute, Pascale Foundation, Naples, Italy. Disclosures: Authors have nothing to disclose with regard to commercial support. Received for publication May 15, 2010; accepted for publication June 28, 2010; available ahead of print Aug 5, 2010. Address for reprints: Gaetano Rocco, MD, FRCSEd, Via Terminio 1 Serino (Avellino) 83028 Italy (E-mail: Gaetano.Rocco@btopenworld.com). J Thorac Cardiovasc Surg 2011;141:839-40 0022-5223/$36.00 Copyright 2011 by The American Association for Thoracic Surgery doi:10.1016/j.jtcvs.2010.06.050