Additional file 10: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1α axis

Figure S5. Representative western blots and corresponding densitograms showing that in K562 (a) and LAMA84 cells (b) curcumin decreased nuclear levels of HIF-1α. Ponceau S of nuclear extract was used as loading control. Intensities of proteins band (in Ponceau S the band used is indicated with arrow) were calculated from the peak area of densitogram by using Image J software. Ctrl: control cells. (PPTX 809 kb)

ROLE OF EXOSOMES RELEASED BY CHRONIC MYLEOGENOUS LEUKEMIA CELLS IN THE MODULATION OF TUMOR MICROENVIROMENT

Shedding of membrane vesicles containing HSP70 and FGF-2 from A6 stem cells.

Analysis of intracellular FGF-2 trafficking in Sk-Hep1 cells.

Vaccine and immune cell therapy in non-small cell lung cancer

Abstract: Despite new advances in therapeutics, lung cancer remains the first cause of mortality among different types of malignancies. To improve survival, different strategies have been developed such as chemotherapy combinations, targeted therapies and more recently immunotherapy. Immunotherapy is based on the capability of the immune system to differentiate cancer cells from normal cells to fight against the tumor. The two main checkpoint inhibitors that have been widely studied in non-small cell lung cancer (NSCLC) are PD-1/PD-L1 and CTLA-4. However, interactions between tumor and immune system are much more complex with several different elements that take part and probably many new i…

Mouse A6 stem cells release active FGF-2 into extracellular space through plasma membrane vesicles

In this study, mouse mesoangioblasts were seeded onto bidimensional matrices within three-dimensional porous scaffolds of poly (L-lactic acid) (PLLA), in the presence or absence of a type I collagen coating. The cells were observed under a scanning electron microscope and tested for their adhesion, survival and proliferation. Immunolocalization of heat shock protein (Hsp) 70, an abundant and ubiquitous intracellular protein in these cells, was also performed in sectioned cell-containing scaffolds under a confocal fluorescence microscope to determine if in situ analysis of intracellular constituents was feasible. The data show that PLLA films allow direct cell adhesion and represent an optim…

Effects of Parietaria judaica on human lung microvascular endothelial cells

SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1α axis

Background Chronic myelogenous leukemia (CML) is a myeloproliferative disorder caused by expression of the chimeric BCR-ABL tyrosine kinase oncogene, resulting from the t(9;22) chromosomal translocation. Imatinib (gleevec, STI-571) is a selective inhibitor of BCR-ABL activity highly effective in the treatment of CML. However, even though almost all CML patients respond to treatment with imatinib or third generation inhibitors, these drugs are not curative and need to be taken indefinitely or until patients become resistant. Therefore, to get a definitive eradication of leukemic cells, it is necessary to find novel therapeutic combinations, for achieving greater efficacy and fewer side effec…

Production of an egg yolk antibody against Parietaria judaica 2 allergen

Specific antibodies are essential tools for studying proteins as well as for diagnostic research in biomedicine. The egg yolk of immunized chicken is an inexpensive source of high-quality polyclonal antibodies. The 12-kDa Parietaria judaica 2 allergen was expressed as a fusion protein and was used to immunize Leghorn chickens. In this paper, we show, using 2-dimensional gel electrophoresis and immunoblotting, that chicken antibodies raised against a recombinant allergen can be used to recognize similar proteins from a pollen raw extract. Allergen identity was confirmed by nanoLC-nanospray-tandem mass spectrometry analysis. Our data demonstrate for the first time that a synergistic combinati…

Can curcumin induces selective packaging of miRNAs in exosomes? A pilot study in chronic myelogenous leukemia cells.

e13563 Background: Chronic myeloid leukemia is a myeloproliferative disorder that comes from a hematopoietic stem cell or a multipotent progenitor. Exosomes are biological nanovescicles (40-100 nm)...

Analysis of FGF-2 traffiking in Sk-Hep1 cells

Additional file 11: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1Îą axis

Figure S6. IPO7/miRNAs correlation. a Analysis performed by using microRNA target prediction software miRSearch V3.0 showed that IPO7 is a validated target of miR-22 and miR-9. b Analysis of predicted multiple targets performed by MicroRNA Target prediction (miRTar) tool ( http://mirtar.mbc.nctu.edu.tw/human/ ) revealed within the CurcuDown-Regulated dataset the presence of several of miR-22 targets beside IPO7. No target of miR-9 was found. (PPTX 179 kb)

carboxyamidotriazole-orotate inhibits the growth of imatinib resistant chronic myeloid leukaemia cells and modulates exosomes-stimulated angiogenesis

The Bcr/Abl kinase has been targeted for the treatment of chronic myelogenous leukaemia (CML) by imatinib mesylate. Although more common in solid tumors, increased microvessel density was also reported in chronic myelogenous leukaemia and was associated with a significant increase of angiogenic factors, suggesting that vascularity in haematologic malignancies is a controlled process and may play a role in the leukaemogenic process thus representing an alternative therapeutic target. Carboxyamidotriazole-orotate (CTO) is the orotate salt form of carboxyamidotriazole (CAI), an orally bioavailable signal transduction inhibitor that in vitro has been shown to possess antileukaemic activities (1…

Membrane vesicles containing matrix metalloproteinase-9 and fibroblast growth factor-2 are released into the extracellular space from mouse mesoangioblast Sstem cells

Certain proteins, including fibroblast growth factor-2 (FGF-2) and matrix metalloproteinase-9 (MMP-9), have proved very effective in increasing the efficacy of mesoangioblast stem cell therapy in repairing damaged tissue. We provide the first evidence that mouse mesoangioblast stem cells release FGF-2 and MMP-9 in their active form through the production of membrane vesicles. These vesicles are produced and turned over continuously, but are stable for some time in the extracellular milieu. Mesoangioblasts shed membrane vesicles even under oxygen tensions that are lower than those typically used for cell culture and more like those of mouse tissues. These findings suggest that mesoangioblast…

Cellule staminali A6 di topo producono vescicole che contengono HSP70 e FGF-2

Curcumin modulates chronic myelogenous leukemia exosomes composition and affects angiogenic phenotype, via exosomal miR-21.

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Proteomic profiling and functional characterization of metastatic colon cancer exosomes spreading malignant properties in tumor microenvironment

Human tumors display a remarkable intratumor heterogeneity affecting clinically relevant phenotypes such as ability to metastasize or to tolerate cytotoxic drugs. Recent published data indicate that tumor derived exosomes (TDEs) can have a pivotal role in regulating tumor heterogeneity by transferring functional biomolecules between various populations of tumor cells and between tumor cells and nontumor cells with consequences for whole tumor microenvironment. In this context, our goal was to understand if exosomes derived from highly metastatic cell line may influence the behaviour of less aggressive tumor cells and the properties of endhothelium.

Localization of neutral ceramidase and sphingosine kinase1 in hepatoma and breast carcinoma cells and their shed vesicles

Targeting of multiple myeloma-related angiogenesis by miR-199a-5p mimics: in vitro and in vivo anti-tumor activity

// Lavinia Raimondi 1 , Nicola Amodio 1 , Maria Teresa Di Martino 1 , Emanuela Altomare 1 , Marzia Leotta 1 , Daniele Caracciolo 1 , Annamaria Gulla 1 , Antonino Neri 2 , Simona Taverna 3 , Patrizia D’Aquila 4 , Riccardo Alessandro 3 , Antonio Giordano 5 , Pierosandro Tagliaferri 1 and Pierfrancesco Tassone 1,5 . 1 Department of Experimental and Clinical Medicine, Magna Graecia University and Medical Oncology Unit, T. Campanella Cancer Center, Salvatore Venuta University Campus, Catanzaro, Italy 2 Department of Medical Sciences University of Milan, Hematology1, IRCCS Policlinico Foundation, Milan, Italy 3 Department of Pathology and Forensic and Medical Biotechnology, Section of Biology and…

Role of miR-126 shuttled by exosomes in the crosstalk between chronic myelogenus leukemia and endothelial cells.

Abstract 4372: Chronic myeloid leukemia (CML) exosomes promote angiogenesis in a Src-dependent fashion in vitro and in vivo

Abstract CML is an uncontrolled proliferation of bone marrow myeloid cells driven by the constitutively active fusion product tyrosine kinase BCR/ABL. Angiogenesis, the formation of new blood vessels from pre-existing vasculature, is newly recognized as a factor in CML progression. Exosomes, released by a broad spectrum of cells, are microvesicles that play an important role in cell-to-cell communication both in physiological and pathological conditions. The role of exosomes released by CML cells in angiogenesis is emerging; however, little is known about the mechanisms involved in this process. We first isolated and characterized exosomes released by K562 CML cells and we demonstrated thei…

Role of microRNAs shuttled by exosomes in the crosstalk between chronic myelogenous leukemia and endothelial cells

Exosome-mediated drug resistance in cancer: the near future is here.

Drug resistance exerts a crucial role in several cancer treatments. Understanding the resistance mechanisms against different therapeutic agents can be helpful to determine the prognosis, but remains a tricky task. In this context, tumor-derived exosomes (TDEs) may give crucial answers about these resistance mechanisms. Exosomes are biological nanovesicles with an average size around 30–100 nm of diameter (Figure 1) that originate from the endocytic pathway by the inward budding of multivesicular bodies (MVB), and they function as cell-free messengers, involved in the cell–cell communication [Kowal et al. 2014]. It has been demonstrated that both cells in physiological and pathological cond…

Comparative analyses of endothelial and tumoral cells cultured in 2D and 3D type-1 collagen fibril gels

Can Be miR-126-3p a Biomarker of Premature Aging? An Ex Vivo and In Vitro Study in Fabry Disease

Fabry disease (FD) is a lysosomal storage disorder (LSD) characterized by lysosomal accumulation of glycosphingolipids in a wide variety of cytotypes, including endothelial cells (ECs). FD patients experience a significantly reduced life expectancy compared to the general population

Additional File 12:

Figure S7. Anti-proliferative effects of curcumin, imatinib and curcumin+imatinib combination on CML cell viability. Curcumin and imatinib were tested for their anti-proliferative effects on K562 (a) and LAMA84 cells (b). The assays were performed by using curcumin and imatinib singly (using the reported doses) or in combination (20 μM curcumin held constant and imatinib at reported concentrations. In K562 cells combination compound treatments showed significant differences compared to single imatinib treatments for all doses tested (p 1 indicates antagonism. (PPTX 50 kb)

CarboxyAmido-Triazole Orotate inhibits the growth of Imatinib-resistant chronic myeloid leukaemia cells and modulates exosomes-stimulated angiogenesis

Chronic myelogenous leukemia is a myeloproliferative disorder characterized by the t(9:22) (q34:q11) reciprocal chromosomal translocation, resulting in the expression of the chimeric Bcr–Abl oncoprotein with constitutive tyrosine kinase activity. Deregulated Bcr–Abl induces the hyperactivation of various signalling pathways that promote cell growth, suppress apoptosis and alter cell adhesion. Bcr-Abl has also been involved in VEGF-mediated angiogenesis in CML and evidence indicates that the formation of new vessels plays an important role in the development and progression of CML. Imatinib mesylate (IM) is a selective well tolerated inhibitor of the Bcr–Abl tyrosine kinase that has signific…

Exosomes from metastatic cancer cells transfer amoeboid phenotype to non-metastatic cells and increase endothelial permeability: their emerging role in tumor heterogeneity

AbstractThe goal of this study was to understand if exosomes derived from high-metastatic cells may influence the behavior of less aggressive cancer cells and the properties of the endothelium. We found that metastatic colon cancer cells are able to transfer their amoeboid phenotype to isogenic primary cancer cells through exosomes, and that this morphological transition is associated with the acquisition of a more aggressive behavior. Moreover, exosomes from the metastatic line (SW620Exos) exhibited higher ability to cause endothelial hyperpermeability than exosomes from the non metastatic line (SW480Exos). SWATH-based quantitative proteomic analysis highlighted that SW620Exos are signific…

P1.17 How to Visualize Exosomes in NSCLC: “The New Guest Star in the Liquid Biopsy Movie”

Additional file 5: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1α axis

Figure S3. Pearson’s R2 showing the correlation between biological and technical replicates of Curcu-K562 cells. (PPTX 185 kb)

An active form of sphingosine kinase-1 is released in the extracellular medium as component of membrane vesicles shed by two human tumor cell line.

Expression of sphingosine kinase-1 (SphK-1) correlates with a poor survival rate of tumor patients. This effect is probably due to the ability of SphK-1 to be released into the extracellular medium where it catalyzes the biosynthesis of sphingosine-1-phosphate (S1P), a signaling molecule endowed with profound proangiogenic effects. SphK-1 is a leaderless protein which is secreted by an unconventional mechanism. In this paper, we will show that in human hepatocarcinoma Sk-Hep1 cells, extracellular signaling is followed by targeting the enzyme to the cell surface and parallels targeting of FGF-2 to the budding vesicles. We will also show that SphK-1 is present in a catalitycally active form i…

Exosomes released by k562 chronic myeloid leukemia cells promote endothelial cell tubular differentiation through uptake and cell-to-cell transfer

Exosomes, microvesicles of endocytic origin released by normal and tumor cells, play an important role in cell-to-cell ommunication. Angiogenesis has been shown to regulate progression of chronic myeloid leukemia (CML). The mechanism through which this happens has not been elucidated. We isolated and characterized exosomes from K562 CML cells and evaluated their effects on human umbilical endothelial cells (HUVECs). Fluorescent-labeled exosomes were nternalized by HUVECs during tubular differentiation on Matrigel. Exosome localization was perinuclear early in differentiation, moving peripherally in cells undergoing elongation and connection. Exosomes move within and between nanotubular stru…

Ectosomes containing HSP70 and FGF-2 are released from mouse A6 stem cells

Primary and metastatic brain cancer genomics and emerging biomarkers for immunomodulatory cancer treatment

Abstract: Recent studies with immunomodulatory agents targeting both cytotoxic T-lymphocyte protein 4 (CTLA4) and programmed cell death 1 (PD1)/programmed cell death ligand 1 (PDL1) have shown to be very effective in several cancers revealing an unexpected great activity in patients with both primary and metastatic brain tumors. Combining anti-CTLA4 and anti-PD1 agents as upfront systemic therapy has revealed to further increase the clinical benefit observed with single agent, even at cost of higher toxicity. Since the brain is an immunological specialized area it's crucial to establish the specific composition of the brain tumors' micro environment in order to predict the potential activit…

In vitro and in vivo effect of CarboxyAmido-Triazole Orotate (CTO) on Imatinib resistant Chronic Myelogenous Leukemia cells

role of CML exosomes in the crosstalk between chronic myelogenous leukaemia and bone marrow-derived cells

Exosomes are small membrane vesicles (30–100 nm) derived from the luminal membranes of multivesicular bodies and constitutively released by fusion with the cell membrane (1). Exosomes mediate local and systemic cell communication through the presence of cytokines, growth factors and others molecules. It is well recognized that bone marrow–derived cells (BMDCs) are crucial for the generation of a suitable microenvironment for the primary tumor and the development of metastasis through a process called pre-metastatic niche formation. Secreted factors are known contributors to BMDC recruitment to both the primary tumor and to pre-metastatic niches (2) and in particular exosomes may have a role…

Shedding of Membrane Vesicles Mediates Fibroblast Growth Factor-2 Release from Cells

Fibroblast growth factor-2 (FGF-2), a polypeptide with regulatory activity on cell growth and differentiation, lacks a conventional secretory signal sequence, and its mechanism of release from cells remains unclear. We characterized the role of extracellular vesicle shedding in FGF-2 release. Viable cells released membrane vesicles in the presence of serum. However, in serum-free medium vesicle shedding was dramatically down-regulated, and the cells did not release FGF-2 activity into their conditioned medium. Addition of serum to serum-starved cells rapidly induced intracellular FGF-2 clustering under the plasma membrane and into granules that colocalized with patches of the cell membrane …

Corrigendum to “Liquid biopsies in lung cancer: The new ambrosia of researchers” [Biochem. Biophys. Act. 1846(2014) 539–546]

a Phase I — Early Clinical Trials Unit, Oncology Department, Antwerp University Hospital, Wilrijkstraat 10, Edegem 2650, Belgium b Molecular Pathology Unit, Pathology Department, Antwerp University Hospital, Wilrijkstraat 10, Edegem 2650, Belgium c Department of Surgical, Oncological and Oral Sciences, Section of Medical Oncology, University of Palermo, Via Liborio Giuffre 5, Palermo 90127, Italy d Department of Investigational Cancer Therapeutics (Phase I Program), The University of Texas MD Anderson Cancer Center, Holcombe Blvd 1400, Unit 455, Houston 77030, USA e Department of Biopathology and Medical and Forensic Biotechnologies, Section of Biology and Genetics, University of Palermo, V…

Circular RNA in Exosomes

Circular RNAs (circRNAs) are a novel family of non-coding endogenous RNAs discovered in all eukaryotic cells and generated through a particular mechanism of alternative splicing called “back-splicing”. These molecules show multiple functions, by acting as modulators of gene and miRNA expression, and may have a role in several biological processes, such as cell proliferation and invasion with, tumour development and progression, and in several mechanisms underlying other diseases. Their presence has been shown to be abundant in several body fluids such as blood and saliva. Based on their biogenesis mechanism, cir- cRNAs may be categorized into five classes: exonic circRNAs, intronic circRNAs…

Tumor microenvironment modulation by exosomes in chronic myelogenous leukemia

Exosomes are small vesicles of 40-100 nm diameter that are initially formed within the endosomal compartment and are secreted when a multivesicular body (MVB) fuses with the plasma membrane. These vesicles are released by many cell types including cancer cells and are considered messengers in intercellular communication. The exact function of exosomes in malignant cells has yet to be elucidated, but investigation has suggested roles in cell-to-cell communication, tumor-stroma interaction, and antigen presentation, thus potentially affecting cancer progression at different steps. Although production of exosomes by CML cells has been reported, little is known regarding the role of these vesic…

miR-126-3p and miR-21-5p as Hallmarks of Bio-Positive Ageing; Correlation Analysis and Machine Learning Prediction in Young to Ultra-Centenarian Sicilian Population

Human ageing can be characterized by a profile of circulating microRNAs (miRNAs), which are potentially predictors of biological age. They can be used as a biomarker of risk for age-related inflammatory outcomes, and senescent endothelial cells (ECs) have emerged as a possible source of circulating miRNAs. In this paper, a panel of four circulating miRNAs including miR-146a-5p, miR-126-3p, miR-21-5p, and miR-181a-5p, involved in several pathways related to inflammation, and ECs senescence that seem to be characteristic of the healthy ageing phenotype. The circulating levels of these miRNAs were determined in 78 healthy subjects aged between 22 to 111 years. Contextually, extracellular miR-1…

Circulating miRNAs in Successful and Unsuccessful Aging. A Mini-review

Aging is a multifactorial process that affects the organisms at genetic, molecular and cellular levels. This process modifies several tissues with a negative impact on cells physiology, tissues and organs functionality, altering their regeneration capacity. The chronic low-grade inflammation typical of aging, defined as inflammaging, is a common biological factor responsible for the decline and beginning of the disease in age. A murine parabiosis model that combines the vascular system of old and young animals, suggests that soluble factors released by young individuals may improve the regenerative potential of old tissue. Therefore, circulating factors have a key role in the induction of …

Additional file 1: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1α axis

Figure S1. Cell growth was measured by MTT assay after 24 h of treatment with increasing doses of curcumin. Each point represents the mean ± SD of three independent experiments. * ≤ 0.05. (PPTX 42 kb)

Curcumin induces selective packaging of miR-21 in exosomes released by Chronic Myelogenous Leukemia cells

Exosomal shuttling of miR-126 in endothelial cells modulates adhesive and migratory abilities of chronic myelogenous leukemia cells.

BACKGROUND: Recent findings indicate that exosomes released from cancer cells contain microRNAs (miRNAs) that may be delivered to cells of tumor microenvironment. RESULTS: To elucidate whether miRNAs secreted from chronic myelogenous leukemia cells (CML) are shuttled into endothelial cells thus affecting their phenotype, we first analysed miRNAs content in LAMA84 exosomes. Among the 124 miRNAs identified in LAMA84 exosomes, we focused our attention on miR-126 which was found to be over-overexpressed in exosomes compared with producing parental cells. Transfection of LAMA84 with Cy3-labelled miR-126 and co-culture of leukemia cells with endothelial cells (EC) confirmed that miR-126 is shuttl…

Contribution of proteomics to understanding the role of tumor-derived exosomes in cancer progression: State of the art and new perspectives

Exosomes are nanometer-sized vesicles (40-100 nm diameter) of endocytic origin released from different cell types under both normal and pathological conditions. They function as cell free messengers, playing a relevant role in the cell-cell communication that is strongly related to the nature of the molecules (proteins, mRNAs, miRNAs, and lipids) that they transport. Tumor cells actively shed exosomes into their surrounding microenvironment and growing evidence indicates that these vesicles have pleiotropic functions in the regulation of tumor progression, promoting immune escape, tumor invasion, neovascularization, and metastasis. During the last few years remarkable efforts have been made…

Effect of exosomes released from K562 CML cell line on gamma-delta T cells function

Application of an immunoproteomic approach to detect anti-profilin antibodies in sera of paritaria judaica allergic patients

Pollen from grasses, weeds, and trees constitutes one of the main sources of inhalant allergens frequently associated with seasonal patterns of allergic diseases. Pollen allergens show some analogies in the amino acids sequence which determine immunological similarity and cross reactivity. Parietaria judaica (P.j) pollen represents one of the main sources of allergens in the Mediterranean area and its major allergens have already been identified (Par j 1 and Par j 2). Recently, has been also described a minor allergen, profilin (Par j 3), an allergen present in pollen of trees, grasses and weeds. Allergenic plant profilins constitute a highly conserved family with sequence identities of 70%…

Detection of RAS mutations in circulating tumor DNA: a new weapon in an old war against colorectal cancer. A systematic review of literature and meta-analysis

Background: Tissue evaluation for RAS (KRAS or NRAS) gene status in metastatic colorectal cancer (mCRC) patients represent the standard of care to establish the optimal therapeutic strategy. Unfortunately, tissue biopsy is hampered by several critical limitations due to its invasiveness, difficulty to access to disease site, patient’s compliance and, more recently, neoplastic tissue spatial and temporal heterogeneity. Methods: The authors performed a systematic literature review to identify available trials with paired matched tissue and ctDNA RAS gene status evaluation. The authors searched EMBASE, MEDLINE, Cochrane, www.ClinicalTrials.gov , and abstracts from international meetings. In to…

Carboxyamidotriazole-Orotate Inhibits the Growth of Imatinib-Resistant Chronic Myeloid Leukaemia Cells and Modulates Exosomes-Stimulated Angiogenesis

The Bcr/Abl kinase has been targeted for the treatment of chronic myelogenous leukaemia (CML) by imatinib mesylate. While imatinib has been extremely effective for chronic phase CML, blast crisis CML are often resistant. New therapeutic options are therefore needed for this fatal disease. Although more common in solid tumors, increased microvessel density was also reported in chronic myelogenous leukaemia and was associated with a significant increase of angiogenic factors, suggesting that vascularity in hematologic malignancies is a controlled process and may play a role in the leukaemogenic process thus representing an alternative therapeutic target. Carboxyamidotriazole-orotate (CTO) is …

Extracellular Vesicles-ceRNAs as Ovarian Cancer Biomarkers: Looking into circRNA-miRNA-mRNA Code

Simple Summary Patients with ovarian cancer have a very poor chance of long-term survival, usually due to advanced disease at the time of diagnosis. Emerging evidence suggests that extracellular vesicles contain noncoding RNAs such as microRNAs, piwiRNAs, circular RNAs, and long noncoding RNAs, with regulatory effects on ovarian cancer. In this review, we focus on ovarian cancer-associated circular RNA shuttled by extracellular vesicles as mediators of cancer progression and novel biomarkers in liquid biopsy. We propose a circular-RNA-microRNA-mRNA code that can reveal the regulatory network created by extracellular vesicles, noncoding RNAs, and mRNAs in ovarian cancer. Future research in t…

Proteomic analysis of Parietaria judaica pollen and allergen profiling by an immunoproteomic approach

Parietaria judaica pollen is a common cause of airway allergic disease in the Mediterranean area. Proteome analysis of mature Parietaria judaica pollen by two-dimensional gel electrophoresis (2-DE) and mass spectrometry has established the first reference proteome map of this weed. Proteins involved in a variety of cellular functions as well as the occurrence of allergens were detected. By using 2-DE and immunoblotting with sera from Parietaria judaica allergic patients we obtained a more detailed characterization of Parietaria judaica allergen profile so to improve our comprehension of the pathogenesis of pollen-induced allergic reaction.

Additional file 9: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1α axis

Figure S4. Effects of Curcumin on HIF-1α activity, IPO7 expression and miR22 expression in LAMA84 cells. a Assay of the transcriptional activity of HIF-1α showing that in LAMA84 cells curcumin induced a reduction of HIF-1α activity compared to control cells. The reported values are the mean of three independent experiments. b qPCR (left panel) and representative Western blot (right panel) show that in LAMA84 cells curcumin treatment did not affect HIF-1α at both mRNA and protein level. The values (FOI: Fold of Induction) in the histogram are normalized against GAPDH and are the mean ± SD of three independent experiments. c qPCR demonstrates that in LAMA84 cells curcumin induced a decrease o…

Effect of Parietaria judaica pollen on human lung microvascular endothelial cells.

Role of exosomes released by chronic myelogenous leukemia cells in angiogenesis

The present study is designed to assess if exosomes released from Chronic Myelogenous Leukemia (CML) cells may modulate angiogenesis. We have isolated and characterized the exosomes generated from LAMA84 CML cells and demonstrated that addition of exosomes to human vascular endothelial cells (HUVEC) induces an increase of both ICAM-1 and VCAM-1 cell adhesion molecules and interleukin-8 expression. The stimulation of cell-cell adhesion molecules was paralleled by a dose-dependent increase of adhesion of CML cells to a HUVEC monolayer. We further showed that the treatment with exosomes from CML cells caused an increase in endothelial cell motility accompanied by a loss of VE-cadherin and β-ca…

Carboxyamidotriazole-orotate inhibits the growth of Imatinib resistantchronic myeloid leukemia cells and modulates exosomes stimulated Angiogenesis

Chronic myeloid leukemia (CML) is characterized by the expression of Bcr–Abl oncoprotein with a constitutive tyrosine kinase that drives disease pathogenesis. Imatinib is the election therapy for CML, but some patients are resistant to this drug. Recently, attention is being focused on cell-cell communication that involves membrane vesicles called exosomes. A number of studies have described exosomes as new players in modulating the tumor microenvironment, promoting angiogenesis and tumor development; furthermore neovascularization is known to exert an important role in the progression of chronic myeloid leukaemia and may represent a valid alternative target for therapy. Little is known reg…

Exosomes released by chronic myelogenous leukemia cells modulate gamma-delta T cell activities

Intracellular trafficking of endogenous fibroblast growth factor‐2

We have previously reported how the release of fibroblast growth factor-2 (FGF-2) is mediated by shed vesicles. In the present study, we address the question of how newly synthesized FGF-2 is targeted to the budding vesicles. Considering that in vitro cultured Sk-Hep1 hepatocarcinoma cells release FGF-2 and shed membrane vesicles only when cultured in the presence of serum, we added serum to starved cells and monitored intracellular movements of the growth factor. FGF-2 was targeted both to the cell periphery and to the nucleus and nucleolus. Movements toward the cell periphery were not influenced by drugs affecting microtubules, but were inhibited by cytocalasin B. Involvement of actin in …

Exosomes in semen: opportunities as a new tool in prostate cancer diagnosis

Abstract: Prostate cancer (PCa) is the most common cancer in men. Nowadays, it is diagnosed through the test of serum prostate-specific antigen (PSA) and rectal examination; however, there is still debate about the PSA-based diagnosis. Seminal fluid (SF), contains a high concentration of subcellular lipid-bound microparticles, traditionally termed "prostasomes", that are extracellular vesicles (EVs) released into the extracellular space by prostate gland's epithelial cells. These vesicles, first described in 1982 promote motility of sperm cells, regulation of sperm cell capacitation, acrosome reaction and immune suppression within the female reproductive tract. It was demonstrated that pros…

Shedding of vesicles from mesoangioblast A6 stem cells

Localisation of neutral ceramidase and sphingosine kinase 1 in hepatoma and breastr carcinoma cells and their shed vesicles

Search_strategy_-_Supplemental_Material_1_final – Supplemental material for Detection of RAS mutations in circulating tumor DNA: a new weapon in an old war against colorectal cancer. A systematic review of literature and meta-analysis

Supplemental material, Search_strategy_-_Supplemental_Material_1_final for Detection of RAS mutations in circulating tumor DNA: a new weapon in an old war against colorectal cancer. A systematic review of literature and meta-analysis by Antonio Galvano, Simona Taverna, Giuseppe Badalamenti, Lorena Incorvaia, Marta Castiglia, Nadia Barraco, Francesco Passiglia, Fabio Fulfaro, Giordano Beretta, Giovanni Duro, Bruno Vincenzi, Pierosandro Tagliaferri, Viviana Bazan and Antonio Russo in Therapeutic Advances in Medical Oncology

Increased expression of interleukin-32 in the inflamed ileum of ankylosing spondylitis patients

Objective. To study the mRNA expression and protein tissue distribution of IL-32 in ileal biopsy specimens from patients with AS. Methods. Quantitative gene expression analysis, by real-time PCR, of IL-32, IL-1b, IL-10, TNF-a and IFN-g was performed on ileal biopsies of 15 AS and 15 Crohn’s disease (CD) patients and 10 healthy subjects (HSs). IL-32 tissue distribution was evaluated by immunohistochemistry. The effect of IL-32 on the production of IL-10 by intestinal epithelial cell lines was also evaluated. Results. In the ileal specimens of patients with AS and intestinal chronic inflammation, significant up-regulation of IL-32 at both the mRNA and protein levels was found as compared with…

Meccanismi non convenzionali di secrezione dell’FGF-2

Exosomes isolation and characterization in serum is feasible in non-small cell lung cancer patients: critical analysis of evidence and potential role in clinical practice

// Simona Taverna 1,2,* , Marco Giallombardo 1,3,* , Ignacio Gil-Bazo 4 , Anna Paola Carreca 3 , Marta Castiglia 3 , Jorge Chacartegui 3 , Antonio Araujo 5 , Riccardo Alessandro 1,2 , Patrick Pauwels 6 , Marc Peeters 7 and Christian Rolfo 3 1 Department of Biopathology and Medical Biotechnology, Section of Biology and Genetics, University of Palermo, Palermo, Italy 2 Institute of Biomedicine and Molecular Immunology (IBIM), National Research Council, Palermo, Italy 3 Phase I-Early Clinical Trials Unit, Oncology Department, Antwerp University Hospital (UZA) and Center for Oncological Research (CORE) Antwerp University, Wilrijkstraat, Edegem, Antwerp, Belgium 4 Department of Oncology, Clinica…

Role of exosomes released by chronic myelogenous leukemia cells in angiogenesis

Improving extracellular vesicles visualization: From static to motion

AbstractIn the last decade, extracellular vesicles (EVs) have become a hot topic. The findings on EVs content and effects have made them a major field of interest in cancer research. EVs, are able to be internalized through integrins expressed in parental cells, in a tissue specific manner, as a key step of cancer progression and pre-metastatic niche formation. However, this specificity might lead to new opportunities in cancer treatment by using EVs as devices for drug delivery. For future applications of EVs in cancer, improved protocols and methods for EVs isolation and visualization are required. Our group has put efforts on developing a protocol able to track the EVs for in vivo intern…

Role of exosomes released by colon cancer stem cells in the modulation of tumor microenvironment.

Curcumin inhibits in vitro and in vivo chronic myelogenous leukemia cells growth : a possible role for exosomal disposal of miR-21

// Simona Taverna 1 , Marco Giallombardo 1 , Marzia Pucci 1 , Anna Flugy 1 , Mauro Manno 2 , Samuele Raccosta 2 , Christian Rolfo 3 , Giacomo De Leo 1 , Riccardo Alessandro 1, 4 1 Dipartimento di Biopatologia e Metodologie Biomediche, Sezione di Biologia e Genetica, Universita di Palermo, Italy 2 Istituto di Biofisica, CNR, Palermo, Italy 3 Phase I - Early Clinical Trials Unit Oncology Department and Center of Oncological Research (CORE), University Hospital Antwerp & Antwerp University, Belgium 4 Istituto di Biomedicina e Immunologia Molecolare (IBIM), Consiglio Nazionale delle Ricerche, Palermo, Italy Correspondence to: Riccardo Alessandro, e-mail: riccardo.alessandro@unipa.it Keywords: e…

Extracellular vesicles: small bricks for tissue repair/regeneration

Extracellular vesicles (EVs) are nano-sized membrane vesicles involved in intercellular communication. EVs have pleiotropic actions in physiological and pathological conditions. The ability of EVs to transports proteins, drugs and nucleic acid, to target specific cells and to increase the stability of therapeutic cargo, make EVs interesting as new devices for the treatment of human disease. In a recently published issue of European journal of pharmaceutical sciences, Silva and colleagues reviewed the ability of EVs to modulate tissue repair and regeneration, focusing on their roles and therapeutic potential as immunomodulatory messengers. In this perspective, we discussed the open questions…

Extracellular vesicles as miRNA nano-shuttles : dual role in tumor progression

[EN] Tumor-derived extracellular vesicles (EVs) have a pleiotropic role in cancer, interacting with target cells of the tumor microenvironment, such as fibroblasts, immune and endothelial cells. EVs can modulate tumor progression, angiogenic switch, metastasis, and immune escape. These vesicles are nano-shuttles containing a wide spectrum of miRNAs that contribute to tumor progression. MiRNAs contained in extracellular vesicles (EV-miRNAs) are disseminated in the extracellular space and are able to influence the expression of target genes with either tumor suppressor or oncogenic functions, depending on both parental and target cells. Metastatic cancer cells can balance their oncogenic pote…

Role of CML exosomes in the crosstalk between chronic myelogenous leukemia and bone-marrow derived exosomes

Natural Compounds: Molecular Weapons against Leukemia’s

Nowadays cancer is one of the main reasons of death all over the world and it is estimated that deaths caused by cancer will grow dramatically in the next decades. Even if chemotherapy is the election therapy for solid tumors, as well as leukemias and lymphomas, cancer treatments are in continuous evolution trying to solve the problem of resistance mainly due to low accumulation of the drug in tumor cells (MDR). Natural compounds represent a valid alternative to treat several disease and recently the scientific community focus on these natural compounds and plant metabolites with therapeutic activities and low toxicities compared with synthetic ones. A combination therapy, that join convent…

Involvement of multiple myeloma cell-derived exosomes in osteoclast differentiation

Bone disease is the most frequent complication in multiple myeloma (MM) resulting in osteolytic lesions, bone pain, hypercalcemia and renal failure. In MM bone disease the perfect balance between bone-resorbing osteoclasts (OCs) and bone-forming osteoblasts (OBs) activity is lost in favour of OCs, thus resulting in skeletal disorders. Since exosomes have been described for their functional role in cancer progression, we here investigate whether MM cell-derived exosomes may be involved in OCs differentiation. We show that MM cells produce exosomes which are actively internalized by Raw264.7 cell line, a cellular model of osteoclast formation. MM cell-derived exosomes positively modulate pre-…

Effetti dell'estratto proteico del polline di Parietaria judaica su cellule endoteliali della microvascolatura polmonare.

Comparative analysis of physical-chemical precipitation methods of circulating exosome isolation from human biofluids.

Curcumin modulates chronic myelogenous leukemia exosomes composition and affects angiogenic phenotype, via exosomal miR-21

Abstract: Tumor derived exosomes are vesicles which contain proteins and microRNAs that mediate cell-cell communication and are involved in angiogenesis and tumor progression. Curcumin derived from the plant Curcuma longa, shows anticancer effects. Exosomes released by CML cells treated with Curcumin contain a high amount of miR-21 that is shuttled into the endothelial cells in a biologically active form. The treatment of HUVECs with CML Curcu-exosomes reduced RhoB expression and negatively modulated endothelial cells motility. We showed that the addition of CML control exosomes to HUVECs caused an increase in IL8 and VCAM1 levels, but Curcu-exosomes reversed these effects thus attenuating …

Effects of Parietaria judaica pollen extract on human microvascular endothelial cells

Abstract Pollinosis from Parietaria judaica is one of the main causes of allergy in the Mediterranean area. The present study is designed to assess if P. judaica pollens contain bioactive compounds able to elicit a functional response in endothelial cells. We have demonstrated that addition of pollen extract to human lung microvascular endothelial cells (HMVEC-L) induces a modification of cell morphology, actin cytoskeletal rearrangements and an increase in endothelial cell permeability. We further showed that the treatment of endothelial cells with pollen extract causes an increase of E-selectin and VCAM-1 protein levels as well as an increase of IL-8 production. The stimulation of cell–ce…

Effects of exosomes released by NSCLC cells on osteoclasts differentiation in ISTITUTO SUPERIORE DI SANITA’, ROMA AICC

Comparative analyses of endothelial and tumoral cells cultured in 2D and 3D type-I collagen fibril gels.

Exosomes as diagnostic and predictive biomarkers in lung cancer

The concept of exosomes has evolved from be considered garbage bags to the demonstration that exosomes could play very interesting roles and functions, from biomarkers detection to the potential of work as drug delivery systems. It has been widely proved that exosomes can contain key molecules important for the tumour development. The current review summarizes the latest investigations developed in the field of predictive exosomal biomarkers. The microRNAs (miRNAs) are the more known molecules due to their amount inside the exosomes and the sensitivity of the techniques available for their study. However, exosomal proteins, RNA and DNA are becoming an interesting and more feasible field of …

Effects of exosomes released by NSCLC cells on osteoclasts differentiation.

Effects of exosomes released by NSCLC cells on osteoclasts differentiation Pucci M., Taverna S., Corrado C., Giallombardo M., Rolfo C. and Alessandro R. Non-small cell lung cancer (NSCLC) has a poorly 5-year survival rate, as a consequence of the delay in the detection of the disease. The majority of patients are diagnosed in an advanced disease stage. Bone metastasis is the most frequent complication in NSCLC resulting in osteolytic lesions. The perfect balance between bone-resorbing osteoclasts (OCs) and bone-forming osteoblasts (OBs) activity is lost in bone metastasis, inducing osteoclastogenesis. Most of the patients with lung cancer are treated with EGFR inhibitors (TKIs). Numerous st…

Additional file 4: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1α axis

Figure S2. Pearson’s R2 showing the correlation between biological and technical replicates of Ctrl-K562 cells. (PPTX 178 kb)

Amphiregulin contained in NSCLC-exosomes induces osteoclast differentiation through the activation of EGFR pathway

AbstractNon-small cell lung cancer (NSCLC) remains the leading cause of cancer-related deaths worldwide. The majority of patients are diagnosed in advanced disease stage. Bone metastasis is the most frequent complication in NSCLC resulting in osteolytic lesions. The perfect balance between bone-resorbing osteoclasts and bone-forming osteoblasts activity is lost in bone metastasis, inducing osteoclastogenesis. In NSCLC, the epidermal growth factor receptor (EGFR) pathway is constitutively activated. EGFR binds Amphiregulin (AREG) that is overexpressed in several cancers such as colon, breast and lung. Its levels in plasma of NSCLC patients correlate with poor prognosis and AREG was recently …

Additional file 7: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1Îą axis

Table S4. UpReg Proteins_FunRichGOterms. (XLSX 35 kb)

Additional file 2: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1Îą axis

Table S1. MS Data of Protein ID. (XLSX 2197 kb)

Additional file 6: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1Îą axis

Table S3. DownReg Proteins_FunRichGOterms. (XLSX 50 kb)

Additional file 3: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1Îą axis

Table S2. SWATH-MS Data. (XLSX 884 kb)

Additional file 8: of SWATH-MS based quantitative proteomics analysis reveals that curcumin alters the metabolic enzyme profile of CML cells by affecting the activity of miR-22/IPO7/HIF-1Îą axis

Table S5. Regulated Proteins_ClueGO Results. (XLSX 22 kb)