The induction of Maspin expression by a glucosamine-derivative has an antiproliferative activity in prostate cancer cell lines

Mammary serine protease inhibitor or Maspin has been characterized as a class II tumor suppressor gene in several cancer types, among them prostate cancer (CaP). Androgen ablation is an effective therapy for CaP, but with short-term effectiveness, thus new therapeutic strategies are actively sought. The present study is aimed to explore the effects of a glucosamine derivative, 2-(N-Carbobenzyloxy)L-phenylalanylamido-2-deoxy-β-D-glucose (NCPA), on two CaP cell lines, PC3 and LNCaP. In particular we analyzed the impact of NCPA on Maspin production, cell viability and cell cycle progression and apoptosis/necrosis pathway activation has been determined in PC3 and LNCaP cell lines. NCPA is able to stimulate Maspin production in PC3 and not in LNCaP cell lines. NCPA blocks the PC3 cell cycle in G1 phase, by inhibiting Cyclin D1 production and induces the apoptosis, therefore interfering with aggressiveness of this androgen-insensitive cell line. Moreover, NCPA is able to induce the expression of Maspin in LNCaP cell line treated with androgen receptor inhibitor, Bicalutamide, and in turn to stimulate the apoptosis of these cells. These findings suggest that NCPA, stimulating the endogenous production of a tumor suppressor protein, could be useful in the design of new therapeutic strategies for treatment of CaP

Case report: The CCDC103 variant causes ultrastructural sperm axonemal defects and total sperm immotility in a professional athlete without primary ciliary diskinesia

Primary ciliary dyskinesia (PCD) is an inherited autosomal-recessive disorder characterized by abnormal ciliary motion, due to a defect in ciliary structure and/or function. This genetic condition leads to recurrent upper and lower respiratory infections, bronchiectasis, laterality defect, and subfertility. Male infertility is often associated with PCD, since the ultrastructure of the axoneme in the sperm tail is similar to that of the motile cilia of respiratory cells. We present the first reported case of a male patient from a non-consanguineous Italian family who exhibited a severe form of asthenozoospermia factor infertility but no situs inversus and absolutely no signs of the clinical respiratory phenotype, the proband being a professional basketball player. Whole-exome sequencing (WES) has identified a homozygote mutation (CCDC103 c.461 A > C, p.His154Pro) in the proband, while his brother was a heterozygous carrier for this mutation. Morphological and ultrastructural analyses of the axoneme in the sperm flagellum demonstrated the complete loss of both the inner and outer dynein arms (IDA and ODA, respectively). Moreover, immunofluorescence of DNAH1, which is used to check the assembly of IDA, and DNAH5, which labels ODA, demonstrated that these complexes are absent along the full length of the flagella in the spermatozoa from the proband, which was consistent with the IDA and ODA defects observed. Noteworthy, TEM analysis of the axoneme from respiratory cilia showed that dynein arms, although either IDAs and/or ODAs seldom missing on some doublets, are still partly present in each observed section. This case reports the total sperm immotility associated with the CCDC103 p.His154Pro mutation in a man with a normal respiratory phenotype and enriches the variant spectrum of ccdc103 variants and the associated clinical phenotypes in PCD, thus improving counseling of patients about their fertility and possible targeted treatments

Effects of a glucosamine-derivative on prostate cancer cell line PC3

Prostate Cancer (CaP) is the most common form of male tumor and is the second leading cause of cancer death. Androgen ablation has proved to be an effective therapy for metastatic prostate cancer, but the regression of metastatic lesions lasts only 18 to 24 months. Mammary serine protease inhibitor or Maspin is a 42 kDa, a non-inhibitory member of the serine protease inhibitor superfamily, and has been characterized as a class II tumor suppressor gene in several cancer types, among them prostate cancer (CaP), due to its ability to inhibit metastasis. In normal prostate epithelial cells, Maspin is highly expressed whereas in prostate cancer cell lines its expression is almost completely suppressed. Previously, it has been demonstrated that NCPA, a glucosamine-derivative synthetized in our laboratory, was able to inhibit IKKα nuclear translocation and to stimulate the production and nuclear localization of Maspin in an osteosarcoma cell line: 143B. IKKα, one of two catalytic subunits of NF-B transcriptional factors, enhances tumor promotion by repressing, among other mechanisms, maspin promoter. Tumor-suppressing and anti-metastatic activities of Maspin have been attributed to its ability to inhibit both invasiveness and cell cycle progression and to stimulate apoptosis of tumor cells. Aim of this presentation is to analyze the ability of NCPA to affect metastatic and proliferation activity of PC3, which is an androgen-insensitive prostate cancer cell line

Glucosamine and its peptidyl-derivative NAPA: novel therapeutic strategy for chondrocytes matrix remodeling

Cartilage degradation, due to an imbalance between anabolic and catabolic rate of chondrocyte metabolism, is the main feature of Osteoarthritis (OA). To date, OA is mainly treated with Non Steroidal Anti-Inflammatory Drugs (NSAIDs), in order to reduce arthritis-related symptoms. In the last decades an increasing number of patients have started to use supplements, such as Glucosamine (GlcN) and chondroitin sulfate, as potential chondroprotective agents. Several in vivo clinical trials as well as in vitro experiments have been performed reporting inconsistent outcomes. Previously, in our lab we analyzed the anabolic effects of GlcN and its N-acetyl-phenylalanine derivative (NAPA) in a rabbit OA model, finding that intra-articular administration of GlcN and NAPA was very effective in reducing cartilage changes in injured rabbit knee. GlcN and NAPA intra-articular administration allows higher concentrations to be reached in the joints compared to oral administration, thus providing an explanation for the ability of both molecules to interfere with OA progression. We also studied the effects of GlcN and NAPA on inflammatory pathways, finding that both molecules can interfere with MAP kinase and NF-kB pathways, by interfering with IKK activity. Finally, we studied the effectiveness of GlcN and NAPA one the biosynthetic activity and hence the matrix production of human primary chondrocytes cultured in micromasses, which represent a good tridimensional culture model. We explored the ability of GlcN and NAPA to stimulate the synthesis of collagen type II (Coll II), Aggrecan (ACAN) and Small Leucine-Rich Proteoglycans (SLRPs). After 6 weeks, micromasses stimulated with GlcN + NAPA still showed a large amount of ECM compared to untreated cells. Moreover, Collagen type II was more abundant and better organized compared to that produced by untreated cells. Finally, cells resulted viable in both treated and untreated micromasses, even if in the middle of untreated micromasses, few dead cells were observed, whereas in the treated micromasses only viable cells and cells completely surrounded by ECM were detected

Analysis of Primary Cilium Expression and Hedgehog Pathway Activation in Mesothelioma Throws Back Its Complex Biology

The primary cilium (PC) is a sensory organelle present on the cell surface, modulating the activity of many pathways. Dysfunctions in the PC lead to different pathologic conditions including cancer. Hedgehog signaling (Hh) is regulated by PC and the loss of its control has been observed in many cancers, including mesothelioma. Malignant pleural mesothelioma (MPM) is a fatal cancer of the pleural membranes with poor therapeutic options. Recently, overexpression of the Hh transcriptional activator GL1 has been demonstrated to be associated with poor overall survival (OS) in MPM. However, unlike other cancers, the response to G-protein-coupled receptor smoothened (SMO)/Hh inhibitors is poor, mainly attributable to the lack of markers for patient stratification. For all these reasons, and in particular for the role of PC in the regulation of Hh, we investigated for the first time the status of PC in MPM tissues, demonstrating intra- and inter-heterogeneity in its expression. We also correlated the presence of PC with the activation of the Hh pathway, providing uncovered evidence of a PC-independent regulation of the Hh signaling in MPM. Our study contributes to the understanding MPM heterogeneity, thus helping to identify patients who might benefit from Hh inhibitors

MYC protein expression scoring and its impact on the prognosis of aggressive B-cell lymphoma patients

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Burkitt lymphoma with a granulomatous reaction: an M1/Th1-polarised microenvironment is associated with controlled growth and spontaneous regression

Aims Burkitt lymphoma (BL) is an aggressive B-cell lymphoma that, in some instances, may show a granulomatous reaction associated with a favourable prognosis and occasional spontaneous regression. In the present study, we aimed to define the tumour microenvironment (TME) in four such cases, two of which regressed spontaneously. Methods and results All cases showed aggregates of tumour cells with the typical morphology, molecular cytogenetics and immunophenotype of BL surrounded by a florid epithelioid granulomatous reaction. All four cases were Epstein–Barr virus (EBV)-positive with type I latency. Investigation of the TME showed similar features in all four cases. The analysis revealed a proinflammatory response triggered by Th1 lymphocytes and M1 polarised macrophages encircling the neoplastic cells with a peculiar topographic distribution. Conclusions Our data provide an in-vivo picture of the role that specific immune cell subsets might play during the early phase of BL, which may be capable of maintaining the tumour in a self-limited state or inducing its regression. These novel results may provide insights into new potential therapeutic avenues in EBV-positive BL patients in the era of cellular immunotherapy