Chronic myelogenous leukaemia exosomes modulate bone marrow microenvironment through activation of epidermal growth factor receptor

Chronic myelogenous leukaemia (CML) is a clonal myeloproliferative disorder. Recent evidence indicates that altered crosstalk between CML and mesenchymal stromal cells may affect leukaemia survival; moreover, vesicles released by both tumour and non-tumour cells into the microenvironment provide a suitable niche for cancer cell growth and survival. We previously demonstrated that leukaemic and stromal cells establish an exosome-mediated bidirectional crosstalk leading to the production of IL8 in stromal cells, thus sustaining the survival of CML cells. Human cell lines used are LAMA84 (CML cells), HS5 (stromal cells) and bone marrow primary stromal cells; gene expression and protein analysis were performed by real-time PCR and Western blot. IL8 and MMP9 secretions were evaluated by ELISA. Exosomes were isolated from CML cells and blood samples of CML patients. Here, we show that LAMA84 and CML patients\u2019 exosomes contain amphiregulin (AREG), thus activating epidermal growth factor receptor (EGFR) signalling in stromal cells. EGFR signalling increases the expression of SNAIL and its targets, MMP9 and IL8. We also demonstrated that pre-treatment of HS5 with LAMA84 exosomes increases the expression of annexin A2 that promotes the adhesion of leukaemic cells to the stromal monolayer, finally supporting the growth and invasiveness of leukaemic cells. Leukaemic and stromal cells establish a bidirectional crosstalk: exosomes promote proliferation and survival of leukaemic cells, both in vitro and in vivo, by inducing IL8 secretion from stromal cells. We propose that this mechanism is activated by a ligand\u2013receptor interaction between AREG, found in CML exosomes, and EGFR in bone marrow stromal cells

IL-17 polarization of MAIT cells is derived from the activation of two different pathways

MAIT cells are expanded in salivary glands of patients with Sjogren's syndrome and are IL-17 polarized. IL-7 and IL-23 induce IL-17 production activating two different pathways: IL-7 stimulation induces in fact a significant STAT3 and HIF1alpha upregulation, conversely, IL-23 stimulation significantly induces RORc overexpression in MAIT cells of patients with Sjogren's syndrome

Multiple myeloma-derived exosomes are enriched of amphiregulin (AREG) and activate the epidermal growth factor pathway in the bone microenvironment leading to osteoclastogenesis

Background: Multiple myeloma (MM) is a clonal plasma cell malignancy associated with osteolytic bone disease. Recently, the role of MM-derived exosomes in the osteoclastogenesis has been demonstrated although the underlying mechanism is still unknown. Since exosomes-derived epidermal growth factor receptor ligands (EGFR) are involved in tumor-associated osteolysis, we hypothesize that the EGFR ligand amphiregulin (AREG) can be delivered by MM-derived exosomes and participate in MM-induced osteoclastogenesis. Methods: Exosomes were isolated from the conditioned medium of MM1.S cell line and from bone marrow (BM) plasma samples of MM patients. The murine cell line RAW264.7 and primary human CD14 + cells were used as osteoclast (OC) sources. Results: We found that AREG was specifically enriched in exosomes from MM samples and that exosomes-derived AREG led to the activation of EGFR in pre-OC, as showed by the increase of mRNA expression of its downstream SNAIL in both RAW264.7 and CD14 + cells. The presence of neutralizing anti-AREG monoclonal antibody (mAb) reverted this effect. Consequently, we showed that the effect of MM-derived exosomes on osteoclast differentiation was inhibited by the pre-treatment of exosomes with anti-AREG mAb. In addition, we demonstrated the ability of MM-derived AREG-enriched exosomes to be internalized into human mesenchymal stromal cells (MSCs) blocking osteoblast (OB) differentiation, increasing MM cell adhesion and the release of the pro-osteoclastogenic cytokine interleukin-8 (IL8). Accordingly, anti-AREG mAb inhibited the release of IL8 by MSCs suggesting that both direct and indirect effects are responsible for AREG-enriched exosomes involvement on MM-induced osteoclastogenesis. Conclusions: In conclusion, our data indicate that AREG is packed into MM-derived exosomes and implicated in OC differentiation through an indirect mechanism mediated by OBs

ANALISI ECONOMICA RETROSPETTIVA SULLE MISURE DI CONTENIMENTO DELLA SPESA FARMACEUTICA DELL’ASP 1 DI AGRIGENTO RELATIVE AL TRIENNIO 2010/2011/2012

Introduzione. Le misure di contenimento della spesa farmaceutica sono state le principali prerogative previste dal Piano di Rientro della Regione Sicilia ( adempimento A.1.2), in tal senso le realtà delle singole ASP si sono adeguate alle direttive regionali mostrando una classifica tra le Asp siciliane che vedono come protagonista virtuosa l’Asp di Agrigento, prima in classifica. La disamina dei dati fa riferimento al triennio 2010/2011/2012. Materiali e metodi. La rilevazione concerne farmaci rimborsati dal SSN ed erogati tramite canale convenzionale privato, farmaci distribuiti direttamente e quelli utilizzati in ambito ospedaliero nel triennio 2010/2011/2012, attraverso report sulla spesa farmaceutica della regione Sicilia. Risultati. Nel 2010 la spesa farmaceutica annua è stata per l’ASP 1 di Agrigento di € 102.942.864,99, questa grazie all’incremento della distribuzione diretta, di cui al PHT D.A.2205/08 attraverso l’introduzione di molecole appartenenti alla categoria N05A (l’11,03 % della spesa totale nell’anno 2011) secondi solo agli antianemici categoria B03X (26,93 % della spesa totale) già presenti nel 2010. La spesa della distribuzione diretta nel 2011 è stata di € 13.973.336,60 mentre la distribuzione attraverso le farmacie private avrebbe fatto lievitare la spesa totale a € 25.006.073,81 nel 2012 invece l’importo è stato di € 14.953.719,04 ed attraverso il canale privato convenzionato avremmo avuto un costo di € 29.530.504,13 . La spesa totale dell’Asp è passata dunque da € 102.942.864,99 nel 2010, a € 92.883.552,99 nel 2011, a € 91.128.889,13 nel 2012. Conclusioni. Le misure adottate per il contenimento della spesa hanno previsto l’introduzione a partire dal 2011 degli antipsicotici atipici, ed il potenziamento del primo ciclo di terapia per il periodo susseguente alla dimissione ospedaliera in base ai criteri dettati dal D.A. 0150/08 portando l’ASP di Ag ad essere tra le più virtuose della regione Sicilia passando dunque dalla sesta posizione in classifica nel 2010 alla prima nel 2012 con un risparmio pari al 11,48 %, il passaggio degli antipsicotici è stato dettato anche dalla necessità dei pazienti che hanno nell’essere sottoposti a controlli periodici da parte degli specialisti ma anche alla valutazione dell’appropriatezza prescrittiva che in quest’ultimo caso è affidato al farmacista ospedaliero. Un altro obiettivo dell’ASP 1 di Agrigento mirato ad un ulteriore contenimento della spesa è quello di estendere la distribuzione diretta anche alla prescrizione successiva ad ogni visita specialistica ambulatoriale

Activated IL-22 pathway occurs in the muscle tissues of patients with polymyositis or dermatomyositis and is correlated with disease activity.

OBJECTIVE: The aim of this study was to assess the expression of IL-22, IL-22 receptor 1 (IL-22R1), IL-22 binding protein (IL-22BP) and p-STAT3 in muscle tissue from patients with PM and DM. METHODS: Levels of IL-22, IL-22R1, IL-22BP and STAT3 mRNA were quantified by RT-PCR. The expression of IL-22, IL-22R1, IL-22BP and p-STAT3 was also analysed using immunohistochemistry. RESULTS: Significant modulation of the IL-22 pathway was observed in inflammatory myopathic tissues. In particular, a significant overexpression of IL-22 at the protein but not the mRNA level was observed in PM/DM tissues and was correlated with myositis activity. IL-22R1 aberrant expression was also observed among infiltrating mononuclear cells and necrotic muscle cells. IL-22BP, which inhibits IL-22 signalling, was expressed only in some muscle fibres in PM/DM patients. CONCLUSION: Our findings indicate that the IL-22 pathway is activated in inflammatory myopathic tissues and may be involved in the induction of muscle inflammatory processes and muscle necrosis

po 146 multiple myeloma derived exosomes carry amphiregulin and are responsible for the uncoupled bone remodelling

Introduction Multiple myeloma (MM) is a hematologic malignancy associated with osteolytic bone disease caused by the perturbation of the functional balance between bone resorption and bone formation. Exosomes, nanosize lipoproteic structures, have been recently recognised as a new mechanism of cell to cell communication during tumour growth and progression. We have previously shown that MM-exosomes are involved in osteolytic lesions but the underlying mechanism is still understood. We hypothesise that the epidermal growth factor receptor ligand Amphiregulin (AREG) can be delivered by multiple myeloma-derived exosomes and participate in modulating the response of the bone microenvironment to the tumour. Material and methods Exosomes were isolated from the conditioned medium of MM1 cell line and from BM plasma samples of patients. In order to test whether MM-exosomes could affect osteoclastogenesis through the activation of the EGFR pathway, primary CD14 +monocytes and a murine cell line (RAW264.7) were used as osteoclast (OC) models. Cells were treated with exosomes from both MM1 and plasma samples, pre-treated or not with anti-AREG neutralising antibodies and OC specific markers were measured. In addition, to further explore whether exosomes were able to promote osteoclastogenesis by affecting mesenchymal stem cells, hTERT-MSC were treated with exosomes; the conditioned medium were collected to measure the secretion of IL8 and to stimulate primary CD14 +monocytes. Results and discussions We found that AREG was specifically enriched in exosome samples, leading to the activation of EGFR in pre-OC. In addition we showed a significant increase of the expression of the OC markers Cathepsin K, Matrix Metalloproteinases 9 and Tartrate-resistant Acid Phosphatase in RAW 264.7 and CD14 +cells after treatment with MM-derived exosomes as compared to the control. The effects of MM-derived exosomes on OC activation were significantly abrogated by exosome pre-treatment with anti-AREG neutralising Ab directly on pre-osteoclast cells and indirectly by inhibiting IL8 release in MSC. Conclusion Taken together our data indicate that MM-derived exosomes are responsible for the uncoupled bone remodelling, affecting directly osteoclast function and promoting IL8 release from mesenchymal stromal cells. In this context, AREG packed into MM-derived exosomes, represents a potential new player in MM-induced bone resorption

Microsatellite instability in gastric cancer is associated with tumor location and family history in a high-risk population from Tuscany

We studied the presence of microsatellite instability (MSI) in a series of 108 gastric cancers (GCs) previously identified in an epidemiological study carried out in a high-risk area around Florence, To investigate associations between MSI and GC family history, 34 cases (31.5%) who had a GC-affected first-degree relative were included in the series, A family history positive for colorectal cancer was reported quite rarely (5.6%). The analysis of 6 microsatellite loci in DNA from paired normal tissue and tumor samples microdissected from paraffin-embedded specimens revealed varying degrees of instability: 56 cases (51.8%) did not show instability at any of the 6 loci; 19 (17.6%) showed instability at 1 locus; 16 (14.8%) showed instability at 2 loci; 11 (10.2%) showed instability at 3 loci; 4 (3.7%) showed instability at 4 loci; and 2 (1.9%) showed instability at 5 loci, The replication error-positive (RER+) phenotype, defined as the presence of MSI at 2 or more loci, had a frequency of 30.6% (33 of 108) and tended to be positively associated with female sex, intestinal histological type, advanced tumor stage, vascular invasion, positive GC family history, and blood group of A type, No correlation emerged between age at diagnosis and RER+ phenotype, whereas a significant association with the RER+ phenotype was shown by the antral location. A multivariate analysis adjusting for a selected group of potential confounding factors confirmed the strong association of the RER+ phenotype with the antral location (P = 0.001) and with a positive GC family history (P < 0.05). Survival analyses at 5 and 8 years showed no difference between RER+ and RER-patients, even when corrected for stage distribution. By the microdissection technique, we also used microsatellite allele patterns to investigate intratumoral heterogeneity and genetic relationships between tumors and adjacent dysplasia and/or intestinal metaplasia. Areas of metaplasia and dysplasia demonstrated MSI only in cases with MSI-positive tumors, In MSI-positive tumors, there was consistent evidence of intratumoral microsatellite allele heterogeneity, indicating the presence of genetically divergent tumor cell clones within the same neoplasm

SULT1A1gene deletion inBRCA2-associated male breast cancer: a link between genes and environmental exposures?

SULT1A1, a member of sulfotransferase superfamily, is a drug and hormone metabolizing enzyme involved in the metabolism of a variety of potential mammary carcinogens of endogenous and exogenous origin. Interestingly, the metabolic activity of SULT1A1 can be affected by varia- tions in gene copy number. Male Breast Cancer (MBC) is a rare disease and less investigated disease compared to female BC (FBC). As in FBC, the concurrent effects of genetic risk factors, particularly BRCA2 mutations, increased exposure to estrogens and environmental carcinogens play a relevant role in MBC. By quantitative real-time PCR with TaqMan probes, we investigated the presence of SULT1A1 gene copy number variations (CNVs) in a series of 72 MBCs. SULT1A1 gene deletion was observed in 10 of the 72 MBCs (13.9%). In a multivariate analysis associ- ation between BRCA2 mutation and SULT1A1 gene deletion emerged (p = 0.0005). Based on the evidence that the level of SULT1A1 enzyme activity is correlated with CNV, our data suggest that in male breast tumors SULT1A1 activity may be decreased. Thus, it can be hypothesized that in a proportion of MBCs, particularly in BRCA2-associated MBCs, the level of estrogens and environmental carcinogens exposure might be increased suggesting a link between gene and environmental exposure in the pathogenesis of MBC

Citrus limon-derived nanovesicles inhibit cancer cell proliferation and suppress CML xenograft growth by inducing TRAIL-mediated cell death

Nanosized vesicles are considered key players in cell to cell communication, thus influencing physiological and pathological processes, including cancer. Nanovesicles have also been found in edible-plants and have shown therapeutic activity in inflammatory bowel diseases; however information on their role in affecting cancer progression is missing.Our study identify for the first time a fraction of vesicles from lemon juice (Citrus limon L.), obtained as a result of different ultracentrifugation, with density ranging from 1,15 to 1,19 g/ml and specific proteomic profile. By using an in vitro approach, we show that isolated nanovesicles inhibit cancer cell proliferation in different tumor cell lines, by activating a TRAIL-mediated apoptotic cell death. Furthermore, we demonstrate that lemon nanovesicles suppress CML tumor growth in vivo by specifically reaching tumor site and by activating TRAIL-mediated apoptotic cell processes. Overall, this study suggests the possible use of plant-edible nanovesicles as a feasible approach in cancer treatment