Crosstalk between Macrophages and Myxoid Liposarcoma Cells Increases Spreading and Invasiveness of Tumor Cells.

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.

Outcome in dedifferentiated chondrosarcoma for patients treated with multimodal therapy: Results from the EUROpean Bone Over 40 Sarcoma Study.

INTRODUCTION: The role of chemotherapy for patients with dedifferentiated chondrosarcoma (DDCS) is still under discussion. Here, we present the outcome in patients with DDCS treated with intensive chemotherapy from the EUROpean Bone Over 40 Sarcoma Study. MATERIALS AND METHODS: The chemotherapy regimen included doxorubicin, ifosfamide and cisplatin. Postoperative methotrexate was added in case of poor histological response. Toxicity was graded based on the National Cancer Institute expanded common toxicity criteria, version 2.0, and survival was analysed using Kaplan-Meier curves, log-rank tests and univariate Cox regression models. RESULTS: Fifty-seven patients with DDCS (localised, 34 [60%]; metastatic, 23 [40%]) aged 42-65 years were included. Surgical complete remission (SCR) was achieved in 36 (63%) patients. The median overall survival (OS) was 24 months (95% confidence interval, 22-25), and the 5-year OS was 39%. Patients with extremity localisation had a 5-year OS of 49% compared with 29% in patients with a central tumour (P = 0.08). Patients with localised disease had a 5-year OS of 46%, whereas patients with metastatic disease had a 5-year OS of 29% (P = 0.12). Patients in SCR had a 5-year OS of 49%, whereas patients not in SCR had a 5-year OS of 23% (P = 0.004). Chemotherapy toxicity was considerable but manageable. There was no treatment-related death, and 39 (70%) patients received ≥6 cycles of the planned nine chemotherapy cycles. CONCLUSIONS: Adding intensive chemotherapy to surgery for treatment of DDCS is feasible and shows favourable survival data compared with previous reports. With the limitations of data from a non-controlled trial, we conclude that chemotherapy could be considered in the management of patients aged >40 years.

Inhibiting Monocyte Recruitment to Prevent the Pro-Tumoral Activity of Tumor-Associated Macrophages in Chondrosarcoma.

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.

Sarcoma Spheroids and Organoids-Promising Tools in the Era of Personalized Medicine.

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.

Intracardiac metastasis originated from chondrosarcoma.

Primary cardiac tumors are extremely rare. By comparison, metastatic involvement of the heart is over 20 times more common and has been reported in autopsy series in up to one in five patients dying of cancer. Cardiac metastasis of chondrosarcoma is absolutely not frequent. In the recent literature, a cardiac metastasis from chondrosarcoma has never been described. We report the case of an 18-year-old man with a diagnosis of cardiac metastasis that originated from a left scapular chondrosarcoma. Chondrosarcoma is a skeletal tumor with various grades of malignancy, rapidly evolving, and with a strong tendency to metastasize, with low responsiveness to chemotherapy. The onset of characteristic systemic symptoms in the late stage of the disease led to the diagnosis of a mass localized in the right atrium. Management and differential diagnosis of infective heart lesions were also very complex in a rapidly evolving life-threatening condition.

Confirmed Activity and Tolerability of Weekly Paclitaxel in the Treatment of Advanced Angiosarcoma.

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/m(2) intravenously, 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.

An unusual association of malignant gastrointestinal neuroectodermal tumor (clear cell sarcoma-like) and Ewing sarcoma.

Very recently a new designation of "Malignant Neuroectodermal Gastrointestinal Tumor" has been proposed for an aggressive form of neuroectodermal tumor with features similar to that of Clear Cell Sarcoma of Soft Tissue, however without a melanocytic differentiation. Also known as "clear cell sarcoma-like tumors of the gastrointestinal tract", these tumors show some features strongly suggesting an origin from a gastrointestinal neuroectodermal precursor cell unable to differentiate along the melanocytic lineage. They occur mainly in young and middle-aged adults, and have a poor prognosis with a high rate of liver and lymphnode metastases. Histologically they are composed of epithelioid or oval-to spindle cells with a sheet-like or nested pattern of growth, strongly positive for neural markers (S-100, SOX10, and vimentin) and negative for the melanocytic ones. EWSR1 gene rearrangements including EWSR1-ATF1 or EWSR1-CREB1 GENE fusions are typically assessed in these tumors. Here we report a case of malignant neuroectodermal gastrointestinal tumor which immunophenotypically unusually expressed FLI-1, occurring in a 29-year-old man with a previous medical history of Ewing sarcoma. We finally suggest that this case might be a further evidence of a link between these two entities.

Anthracycline-based chemotherapy in extraskeletal myxoid chondrosarcoma: a retrospective study.

BACKGROUND: Extraskeletal myxoid chondrosarcoma (EMC) is a rare subgroup within soft tissue sarcomas. Its sensitivity to chemotherapy is reported to be low. METHODS: We retrospectively reviewed a series of 11 EMC patients treated as from 2001 within the Italian Rare Cancer Network (RCN) with anthracycline-based chemotherapy. Pathologic diagnosis was centrally reviewed in all cases and confirmed by the presence of the specific chromosomal rearrangements, involving the NR4A3 gene locus on chromosome 9. RESULTS: Eleven patients treated with anthracycline-based chemotherapy were included (M/F: 9/2 - mean age: 52 years - site of primary: lower limb/other = 9/2 - metastatic = 11 - front line/ further line = 10/1 - anthracycline as single agent/ combined with ifosfamide = 1/10). Ten patients are evaluable for response. Overall, best response according to RECIST was: partial response (PR) = 4 (40 %), stable disease (SD) = 3, progressive disease (PD) = 3 cases. Median PFS was 8 (range 2-10) months. CONCLUSIONS: By contrast to what reported so far, anthracycline-based chemotherapy is active in a distinct proportion of EMC patients.

Treatment of cutaneous angiosarcoma of the face: efficacy of combined chemotherapy and radiotherapy.

INTRODUCTION: Cutaneous angiosarcoma (AS) is a rare form of soft tissue sarcoma. It is aggressive and has a poor prognosis. The aim of our report is to show that with combined chemotherapy and radiotherapy it is possible to obtain good results in terms of local control, complete response, and aesthetic outcome. CASE REPORT: We present the case of a 60-year-old man affected by AS covering the entire surface of the nose. Surgery, although indicated, was excluded because it was considered mutilating and would give a poor cosmetic result. The patient was treated with chemotherapy consisting of paclitaxel 80 mg/m2 for 6 cycles followed by radiotherapy at a dose of 50 Gy. Then 3 additional cycles of chemotherapy were administered according to the same scheme. A complete response was obtained. At 40 months after treatment, the patient did not show any signs of late toxicity, all lesions had disappeared, and all laboratory tests were negative. CONCLUSIONS: Our experience shows that concomitant chemoradiotherapy can be delivered safely and can be tolerated with low toxicity and good results in terms of local control and complete response. We obtained an excellent aesthetic result with improvement of the patient's quality of life.

Ex vivo behaviour of human bone tumor endothelial cells.

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.

Hyperexpression of HOXC13, located in the 12q13 chromosomal region, in well­differentiated and dedifferentiated human liposarcomas.

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 well­differentiated and dedifferentiated LPSs, possibly related to 12q13-15 chromosomal amplification.

Platelet gel in cutaneous radiation dermatitis.

INTRODUCTION: Radiotherapy, alone or in combination with chemotherapy and/or surgery, is a fundamental and irreplaceable method of treating tumours. Nonetheless, although the technological advances made during recent years and the associated improvements in this type of treatment have reduced the incidence of complications, 5-15 % of patients still experience damage to the healthy tissues exposed to radiation. Cutaneous and mucosal lesions are severe collateral effects of radiotherapy that have an enormous impact on a patient's quality of life. Unfortunately, however, the efficacy of conventional treatments, while demonstrably useful in acute lesions, remains disputed in chronic cases. Nevertheless, numerous studies and clinical findings have demonstrated that topical, non-transfusional plasma-rich platelet gel is able to accelerate the regeneration and repair of tissues through the action of the various growth factors contained within the alpha granules of platelets. We therefore set out to evaluate the efficacy of autologous platelet gel, chosen for its limited cost and ease of preparation, in chronic cutaneous radiation dermatitis. METHODS: "Home-made" platelet gel was produced by treating platelets with autologous thrombin. The safety of the product was ensured by microbiological tests. The autologous platelet gel was applied topically once a week, for a mean duration of 35 days, to chronic third- and fourth-degree (European Pressure Ulcer Advisory Panel classification and Common Terminology Criteria for Adverse Events score) cutaneous radiation dermatitis in a group of ten patients previously treated for moderate-to-high grade (histology G2-G3) limb sarcoma by tumour excision and post-surgical radiotherapy (dose 50-64 Gy). The radiation dermatitis had appeared at different intervals after treatment and had all proved resistant to conventional treatments. RESULTS: The autologous platelet gel was found to be successful in seven out of the ten patients treated. The various phases of the healing process were observed in all cases. Platelet gel application was suspended in three patients: in one patient after one application due to tumour progression, in another patient after two applications due to development of distant metastases and in the third after six applications with only partial tissue response. At 5-year follow-up, six of the seven successfully treated patients remained free of both disease and lesion, while the remaining patient, the eldest, had passed away in the interim due to extraneous causes. CONCLUSION: Platelet gel treatment could therefore be used to bring about healing in chronic cutaneous radiation dermatitis, lending itself to better patient compliance and a favourable cost/benefit ratio, due to a reduction in the number of medications and hospital visits required.

Fine-needle cytology of Kaposi's sarcoma in an intramammary lymphnode: report of one case.

Kaposi's sarcoma (KS) is the most common human immunodeficiency virus (HIV) infection disease-associated malignancy. It consists of an angiosarcomatous change of the epithelial and mucous membrane-associated connective tissue not only in various sites, for example, skin, gastrointestinal system, lungs, and so on, but may also involve nonepithelial organs, such as lymphnodes. An unusual localization of KS to an intramammary lymphnode is reported here. The patient, an HIV-negative 69-year-old woman with a clinical history of rheumatoid arthritis treated with hydrocortisone, had an 8-month pathological history of biopsy-proven Kaposi sarcoma of the skin with visceral extension (stomach and duodenum). The appearance of a well-defined 23 × 20 mm(2) breast nodule during chemotherapy elicited fine-needle cytology to exclude breast carcinoma. Surgical excision confirmed the cytopathological diagnosis of Kaposis's sarcoma.

YY1 overexpression is associated with poor prognosis and metastasis-free survival in patients suffering osteosarcoma.

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.

The use of veritas collagen matrix to reconstruct the posterior chest wall after costovertebrectomy.

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.

Involvement of the soluble urokinase receptor in chondrosarcoma cell mobilization.

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 uPAR(88-92) sequence, since the DII(88-183) or DIIDIIR(88-284) uPAR domains retain motogen effect whereas DI(1-87) or DIII(184-284) domains, both lacking the uPAR(88-92) sequence, are ineffective. Chondrosarcoma cells cross matrigel in response to SuPAR, and their invasion capability is abrogated by RERF peptide which inhibits uPAR(88-92) signalling. 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.

Axial chordoma and parachordoma (soft tissue chordoma): two of a kind: report of two cases with primary diagnosis on fine-needle cytology samples.

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.

Primary giant clear cell sarcoma (soft tissue malignant melanoma) of the sternum.

One case of a primary clear cell sarcoma of the sternum (also called soft tissue melanoma) is reported. This neoplasm represents a rare occurrence, and as a rule, differential diagnosis with melanoma often requires detailed immunohistochemistry and cytogenetic analysis (ie, rearrangement of EWS gene localized on 22q12 chromosome). Because wide resection is recommended, chest wall reconstruction may pose challenging technical issues. In our patient, we elected not to proceed to clavicular stabilization. Nevertheless, acceptable shoulder girdle mobility was observed after surgery.

Cell invasiveness in sarcomas: a possibly useful clinical correlation.

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.