82 results on '"Poletto, V."'
Search Results
2. JAK2V617F allele burden ⩾50% is associated with response to ruxolitinib in persons with MPN-associated myelofibrosis and splenomegaly requiring therapy
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Barosi, G, Klersy, C, Villani, L, Bonetti, E, Catarsi, P, Poletto, V, Campanelli, R, Impera, S, Latagliata, R, Viarengo, G, Carolei, A, Massa, M, Musso, M, Crescimanno, A, Gale, R P, and Rosti, V
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- 2016
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3. Functional and genetic aberrations of in vitro-cultured marrow-derived mesenchymal stromal cells of patients with classical Philadelphia-negative myeloproliferative neoplasms
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Avanzini, M A, Bernardo, M E, Novara, F, Mantelli, M, Poletto, V, Villani, L, Lenta, E, Ingo, D M, Achille, V, Bonetti, E, Massa, M, Campanelli, R, Fois, G, Catarsi, P, Gale, R P, Moretta, A, Aronica, A, Maccario, R, Acquafredda, G, Lisini, D, Zecca, M, Zuffardi, O, Locatelli, F, Barosi, G, and Rosti, V
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- 2014
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4. Automotive Electronics
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Morelli, M., primary, Maggioni, G., additional, and Poletto, V., additional
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- 2002
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5. Abnormal expression patterns of WT1-as, MEG3 and ANRIL long non-coding RNAs in CD34+ cells from patients with primary myelofibrosis and their clinical correlations
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Pennucci, V, Zini, R, Norfo, R, Guglielmelli, P, Bianchi, E, Salati, S, Sacchi, G, Prudente, Z, Tenedini, E, Ruberti, S, Paoli, C, Fanelli, T, Mannarelli, C, Tagliafico, E, Ferrari, S, Vannucchi, Am, Manfredini, R, on behalf of Associazione Italiana per la Ricerca sul Cancro Gruppo Italiano Malattie Mieloproliferative Investigators Collaborators Vannucchi AM, Balliu, M, Bartalucci, N, Bogani, C, Bosi, A, Fjerza, R, Martinelli, S, Pancrazzi, A, Pieri, L, Bisognin, Andrea, Bortoluzzi, Stefania, Coppe, Alessandro, Saccoman, Claudia, Masciulli, A, Giovani, B, Azzan, C, Badalucco, S, Balduini, A, Bonetti, E, Campanelli, R, Catarsi, P, Isgrò, Ma, Lupo, Ml, Magrini, U, Massa, M, Poletto, V, Rosti, V, Villani, L, Cazzola, M, Ambaglio, I, Bernasconi, P, Casetti, Ic, Catricalà, S, Elena, C, Fugazza, E, Gallì, A, Malcovati, L, Milanesi, C, Pascutto, C, Pietra, D, Ripamonti, F, Rossi, M, Rumi, E, Dejana, E, Breviario, F, Corada, M, Erba, Bg, Rambaldi, A, Barbui, T, Ferrari, Ml, Finazzi, G, Finazzi, Mc, Gritti, G, Belotti, C, Boroni, C, Salmoiraghi, S, Amaru, A, Golay, J, Cilloni, D, Campia, V, Carturan, S, Guerrasio, A, Montanari, M, and Zini, R.
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medicine.medical_specialty ,Cancer Research ,Antigens, CD34 ,Biology ,Transcriptome ,Internal medicine ,Hematology ,Oncology ,Gene expression ,medicine ,Humans ,Myelofibrosis ,MEG3 ,Regulation of gene expression ,Reverse Transcriptase Polymerase Chain Reaction ,RNA ,Prognosis ,medicine.disease ,Gene Expression Regulation, Neoplastic ,Primary Myelofibrosis ,Cancer cell ,Immunology ,Cancer research ,RNA, Long Noncoding - Abstract
Long non-coding RNAs (lncRNAs), an emerging group of gene expression regulators in normal and cancer cells, are defined as endogenous RNAs consisting of more than 200 nucleotides and lacking an ope...
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- 2015
6. Clinical effect of driver mutations of JAK2, CALR, or MPL in primary myelofibrosis
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Rumi, E, Pietra, D, Pascutto, C, Guglielmelli, P, Martínez Trillos, A, Casetti, I, Colomer, D, Pieri, L, Pratcorona, M, Rotunno, G, Sant'Antonio, E, Bellini, M, Cavalloni, C, Mannarelli, C, Milanesi, C, Boveri, E, Ferretti, V, Astori, C, Rosti, V, Cervantes, F, Barosi, G, Vannucchi, Am, Cazzola, M, Associazione Italiana per la Ricerca sul Cancro Gruppo Italiano Malattie Mieloproliferative Investigators Collaborators Vannucchi AM, Balliu, M, Bartalucci, N, Biamonte, F, Bisognin, A, Bogani, C, Bortoluzzi, S, Bosi, A, Coppe, A, Fanelli, T, Fjerza, R, Loiacono, I, Marchioli, R, Martinelli, S, Masciulli, A, Pancrazzi, A, Paoli, C, Saccoman, C, Spolverini, A, Susini, Mc, Tozzi, L, Azzan, C, Badalucco, S, Balduini, A, Bonetti, E, Campanelli, R, Catarsi, P, Isgrò, Am, Lupo, Ml, Magrini, U, Massa, M, Poletto, V, Villani, L, Ambaglio, I, Bernasconi, P, Casetti, Ic, Catricalà, S, Elena, C, Fugazza, E, Gall, A, Malcovati, L, Ripamonti, F, Rossi, M, Dejana, E, Breviario, F, Corada, M, Erba, Bg, Rambaldi, A, Amaru, A, Barbui, T, Belotti, C, Boroni, C, Ferrari, Ml, Finazzi, G, Finazzi, Mc, Golay, J, Gritti, G, Salmoiraghi, S, Cilloni, Daniela, Campia, V, Carturan, S, Guerrasio, Angelo, Manfredini, R, Bianchi, E, Salati, S, Tagliafico, E, Tenedini, E, and Zini, R.
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Oncology ,Male ,Clinical Trials and Observations ,Leukocytosis ,DNA Mutational Analysis ,Kaplan-Meier Estimate ,Biochemistry ,Risk Factors ,hemic and lymphatic diseases ,Aged, 80 and over ,Leukemia ,biology ,Incidence (epidemiology) ,food and beverages ,Anemia ,Hematology ,Middle Aged ,Prognosis ,Cell Transformation, Neoplastic ,Female ,medicine.symptom ,Receptors, Thrombopoietin ,Adult ,medicine.medical_specialty ,Adolescent ,Immunology ,Lower risk ,Risk Assessment ,Young Adult ,Internal medicine ,medicine ,Humans ,Myelofibrosis ,Aged ,Proportional Hazards Models ,business.industry ,Proportional hazards model ,Cell Biology ,Janus Kinase 2 ,medicine.disease ,Thrombocytopenia ,Primary Myelofibrosis ,Mutation ,biology.protein ,business ,Calreticulin - Abstract
We studied the impact of driver mutations of JAK2, CALR, (calreticulin gene) or MPL on clinical course, leukemic transformation, and survival of patients with primary myelofibrosis (PMF). Of the 617 subjects studied, 399 (64.7%) carried JAK2 (V617F), 140 (22.7%) had a CALR exon 9 indel, 25 (4.0%) carried an MPL (W515) mutation, and 53 (8.6%) had nonmutated JAK2, CALR, and MPL (so-called triple-negative PMF). Patients with CALR mutation had a lower risk of developing anemia, thrombocytopenia, and marked leukocytosis compared with other subtypes. They also had a lower risk of thrombosis compared with patients carrying JAK2 (V617F). At the opposite, triple-negative patients had higher incidence of leukemic transformation compared with either CALR-mutant or JAK2-mutant patients. Median overall survival was 17.7 years in CALR-mutant, 9.2 years in JAK2-mutant, 9.1 years in MPL-mutant, and 3.2 years in triple-negative patients. In multivariate analysis corrected for age, CALR-mutant patients had better overall survival than either JAK2-mutant or triple-negative patients. The impact of genetic lesions on survival was independent of current prognostic scoring systems. These observations indicate that driver mutations define distinct disease entities within PMF. Accounting for them is not only relevant to clinical decision-making, but should also be considered in designing clinical trials.
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- 2014
7. Fine structural detection of calcium ions by photoconversion
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Poletto, V., primary, Galimberti, V., additional, Guerra, G., additional, Rosti, V., additional, Moccia, F., additional, and Biggiogera, M., additional
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- 2016
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8. Acidic Ca2+ stores interact with the endoplasmic reticulum to shape intracellular Ca2+ signals in human endothelial progenitor cells
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Zuccolo, E., primary, Lim, D., additional, Poletto, V., additional, Guerra, G., additional, Tanzi, F., additional, Rosti, V., additional, and Moccia, F., additional
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- 2015
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9. Arachidonic acid stimulates endothelial progenitor cell proliferation through an increase in Ca2+ concentration and nitric oxide production
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Zuccolo, E., primary, Poletto, V., additional, Guerra, G., additional, Rosti, V., additional, and Moccia, F., additional
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- 2015
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10. Tie2 Expressing Monocytes in the Spleen of Patients with Primary Myelofibrosis
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Campanelli, R., Fois, G., Catarsi, P., Poletto, V., Villani, L., Erba, B. G., Maddaluno, L., Jemos, B., Salmoiraghi, S., Guglielmelli, P., Abbonante, V., Di Buduo, C. A., Balduini, A., Iurlo, A., Barosi, G., Rosti, V., Massa, M., Vannucchi, A. M., Balliu, M., Bartalucci, N., Bogani, C., Bosi, A., Calabresi, L., Corbizzi Fattori, G., Fanelli, T., Fjerza, R., Gesullo, F., Mannarelli, C., Merli, L., Pacilli, A., Pancrazzi, A., Paoli, C., Pieri, L., Rotunno, G., Sant'Antonio, E., Bonetti, E., Cazzola, M., Ambaglio, I., Bernasconi, P., Casetti, C. I., Catricala, S., Elena, C., Fugazza, E., Galli, A., Malcovati, L., Milanesi, C., Pascutto, C., Pietra, D., Ripamonti, F., Rossi, M., Rumi, E., Dejana, E., Breviario, F., Corada, M., Malinverno, M., Rambaldi, A., Chioda, G., Ferrari, M. L., Finazzi, G., Finazzi, M. C., Belotti, C., Boroni, C., Amaru, A., Golay, J., Bortoluzzi, S., Bisognin, A., Coppe, A., Saccoman, C., Manfredini, R., Artuso, L., Bernardis, I., Bianchi, E., Montanari, M., Pennucci, V., Prudente, Z., Rontauroli, S., Rossi, C., Ruberti, S., Salati, S., Tagliafico, E., Tenedini, E., and Zini, R.
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Male ,0301 basic medicine ,Pathology ,Physiology ,Angiogenesis ,CD34 ,Gene Expression ,lcsh:Medicine ,Medicine (all) ,Biochemistry, Genetics and Molecular Biology (all) ,Agricultural and Biological Sciences (all) ,Cardiovascular Physiology ,Monocytes ,White Blood Cells ,0302 clinical medicine ,Animal Cells ,Immune Physiology ,Medicine and Health Sciences ,Blood and Lymphatic System Procedures ,Electron Microscopy ,lcsh:Science ,Microscopy ,Multidisciplinary ,Neovascularization, Pathologic ,Cell Differentiation ,Hematology ,Middle Aged ,Receptor, TIE-2 ,Haematopoiesis ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Splenectomy ,cardiovascular system ,Female ,Cellular Types ,Receptor ,Research Article ,medicine.medical_specialty ,Aged ,Case-Control Studies ,Humans ,Primary Myelofibrosis ,Spleen ,Patients ,Immune Cells ,CD14 ,Immunology ,Surgical and Invasive Medical Procedures ,Biology ,Research and Analysis Methods ,03 medical and health sciences ,Genetics ,medicine ,Progenitor cell ,TIE-2 ,Myelofibrosis ,Neovascularization ,Pathologic ,Blood Cells ,lcsh:R ,Biology and Life Sciences ,Cell Biology ,medicine.disease ,Hematopoiesis ,Health Care ,030104 developmental biology ,Transmission Electron Microscopy ,lcsh:Q ,Bone marrow ,Developmental Biology - Abstract
Primary myelofibrosis (PMF) is a Philadelphia-negative (Ph-) myeloproliferative disorder, showing abnormal CD34+ progenitor cell trafficking, splenomegaly, marrow fibrosis leading to extensive extramedullary haematopoiesis, and abnormal neoangiogenesis in either the bone marrow or the spleen. Monocytes expressing the angiopoietin-2 receptor (Tie2) have been shown to support abnormal angiogenic processes in solid tumors through a paracrine action that takes place in proximity to the vessels. In this study we investigated the frequency of Tie2 expressing monocytes in the spleen tissue samples of patients with PMF, and healthy subjects (CTRLs), and evaluated their possible role in favouring spleen angiogenesis. We show by confocal microscopy that in the spleen tissue of patients with PMF, but not of CTRLs, the most of the CD14+ cells are Tie2+ and are close to vessels; by flow cytometry, we found that Tie2 expressing monocytes were Tie2+CD14lowCD16brightCDL62-CCR2- (TEMs) and their frequency was higher (p = 0.008) in spleen tissue-derived mononuclear cells (MNCs) of patients with PMF than in spleen tissue-derived MNCs from CTRLs undergoing splenectomy for abdominal trauma. By in vitro angiogenesis assay we evidenced that conditioned medium of immunomagnetically selected spleen tissue derived CD14+ cells of patients with PMF induced a denser tube like net than that of CTRLs; in addition, CD14+Tie2+ cells sorted from spleen tissue derived single cell suspension of patients with PMF show a higher expression of genes involved in angiogenesis than that found in CTRLs. Our results document the enrichment of Tie2+ monocytes expressing angiogenic genes in the spleen of patients with PMF, suggesting a role for these cells in starting/maintaining the pathological angiogenesis in this organ.
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- 2016
11. Phylogeny of horse chromosome 5q in the genus Equus and centromere repositioning
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Piras, F.M., Nergadze, S.G., Poletto, V., Cerutti, F., Ryder, O.A., Leeb, Tosso, Raimondi, E., and Giulotto, E.
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630 Agriculture ,590 Animals (Zoology) - Abstract
Horses, asses and zebras belong to the genus Equus and are the only extant species of the family Equidae in the order Perissodactyla. In a previous work we demonstrated that a key factor in the rapid karyotypic evolution of this genus was evolutionary centromere repositioning, that is, the shift of the centromeric function to a new position without alteration of the order of markers along the chromosome. In search of previously undiscovered evolutionarily new centromeres, we traced the phylogeny of horse chromosome 5, analyzing the order of BAC markers, derived from a horse genomic library, in 7 Equus species (E. caballus, E. hemionus onager, E. kiang, E. asinus, E. grevyi, E. burchelli and E. zebra hartmannae). This analysis showed that repositioned centromeres are present in E. asinus (domestic donkey, EAS) chromosome 16 and in E. burchelli (Burchell's zebra, EBU) chromosome 17, confirming that centromere repositioning is a strikingly frequent phenomenon in this genus. The observation that the neocentromeres in EAS16 and EBU17 are in the same chromosomal position suggests that they may derive from the same event and therefore, E. asinus and E. burchelli may be more closely related than previously proposed; alternatively, 2 centromere repositioning events, involving the same chromosomal region, may have occurred independently in different lineages, pointing to the possible existence of hot spots for neocentromere formation. Our comparative analysis also showed that, while E. caballus chromosome 5 seems to represent the ancestral configuration, centric fission followed by independent fusion events gave rise to 3 different submetacentric chromosomes in other Equus lineages.
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- 2009
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12. No association between the XPD Lys751Gln (rs13181) polymorphism and disease phenotype or leukemic transformation in primary myelofibrosis
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Poletto, V., primary, Villani, L., additional, Catarsi, P., additional, Campanelli, R., additional, Massa, M., additional, Vannucchi, A. M., additional, Rosti, V., additional, and Barosi, G., additional
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- 2013
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13. Acidic Ca2 + stores interact with the endoplasmic reticulum to shape intracellular Ca2 + signals in human endothelial progenitor cells
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Zuccolo, E., Lim, D., Poletto, V., Guerra, G., Tanzi, F., Rosti, V., and Moccia, F.
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- 2015
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14. Arachidonic acid stimulates endothelial progenitor cell proliferation through an increase in Ca2 + concentration and nitric oxide production
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Zuccolo, E., Poletto, V., Guerra, G., Rosti, V., and Moccia, F.
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- 2015
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15. Phylogeny of Horse Chromosome 5q in the Genus Equus and Centromere Repositioning
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Piras, F.M., primary, Nergadze, S.G., additional, Poletto, V., additional, Cerutti, F., additional, Ryder, O.A., additional, Leeb, T., additional, Raimondi, E., additional, and Giulotto, E., additional
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- 2009
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16. System chips for automotive applications.
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Foletto, A. and Poletto, V.
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- 2004
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17. A R-evolution in power electronics: from "intelligent" to "super smart power" in automotive.
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Marchio, F., Poletto, V., Russo, A., Torrisi, G., Notaro, J., Burlak, G., and Mirowski, M.
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- 2004
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18. JAK2V617Fallele burden ?50% is associated with response to ruxolitinib in persons with MPN-associated myelofibrosis and splenomegaly requiring therapy
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Barosi, G, Klersy, C, Villani, L, Bonetti, E, Catarsi, P, Poletto, V, Campanelli, R, Impera, S, Latagliata, R, Viarengo, G, Carolei, A, Massa, M, Musso, M, Crescimanno, A, Gale, R P, and Rosti, V
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- 2016
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19. Multifunction Voltage Regulator for the automotive environment.
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Poletto, V., Morelli, M., and Mazzucco, M.
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- 1991
20. Wide common mode input range, gain tunable sense amplifier with system self-calibration.
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Weili Zhu, Poletto, V., and Galllinari, M.
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- 2010
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21. T REGULATORY CELLS IN PERIPHERAL BLOOD OF PATIENTS WITH PRIMARY MYELOFIBROSIS
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Campanelli, R., Fois, G., Poletto, V., Villani, L., Paolo Catarsi, Bonetti, E., Rosti, V., Barosi, G., and Massa, M.
22. INCREASED PLASMA NICOTINAMIDE PHOSPHORIBOSYLTRANSFERASE (NAMPT) IS ASSOCIATED WITH A HYPERPROLIFERATIVE PHENOTYPE AND RESTRAINS DISEASE PROGRESSION IN MPN-ASSOCIATED MYELOFIBROSIS
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Barosi, G., Genazzani, A., Travelli, C., Grolla, A., Magrini, U., Fois, G., Catarsi, P., Bonetti, E., Poletto, V., Villani, L., Viarengo, G., Massa, M., Campanelli, R., and Vittorio Rosti
23. CALCIUM INFLUX IS SHAPED BY THE MUTATIONAL STATUS IN CIRCULATING CD34+CELLS OF PATIENTS WITH MYELOFIBROSIS
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Barosi, G., Moccia, F., Tanzi, F., Torti, M., Fois, G., Campanelli, R., Massa, M., Bonetti, E., Villani, L., Canobbio, I., Poletto, V., and Vittorio Rosti
24. VEGF-INDUCED CA2+SIGNALS ARE DERANGED IN ENDOTHELIAL PROGENITOR CELLS FROM PATIENTS WITH MYELOFIBROSIS CARRYING JAK2 MUTATIONS
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Dragoni, S., Poletto, V., FEDERICO ALESSANDRO RUFFINATTI, Barosi, G., Tanzi, F., Rosti, V., and Moccia, F.
25. REDUCED FREQUENCY OF CIRCULATING CD4+CD25+CD127LOWFOXP3+REGULATORY T CELLS IN PRIMARY MYELOFIBROSIS
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Rita Campanelli, Fois, G., Villani, L., Catarsi, P., Bonetti, E., Poletto, V., Viarengo, G., Rosti, V., Gale, R. P., Barosi, G., and Massa, M.
26. HEMATOPOIETIC PROGENITOR CELLS EXPRESSING VE-CADHERIN ARE DETECTABLE IN THE SPLEEN OF PATIENTS WITH MYELOFIBROSIS AND ARE IN CLOSE CONTACT WITH MICROVESSELS
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Campanelli, R., Fois, G., Erba, B. G., Poletto, V., Bonetti, E., Cobianchi, L., Ingo, D., Laura Villani, Catarsi, P., Massa, M., Rosti, V., Dejana, E., and Barosi, G.
27. INTRACELLULAR CALCIUM DEPOSITS AND STORE OPERATED CALCIUM ENTRY IN CD34+CELLS FROM PATIENTS WITH MYELOFIBROSIS CARRYING A CALR MUTATION
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Poletto, V., Canobbio, I., Pietra, D., Dragoni, S., Bonetti, E., Rita Campanelli, Massa, M., Fois, G., Catarsi, P., Villani, L., Torti, M., Tanzi, F., Cazzola, M., Moccia, F., Barosi, G., and Rosti, V.
28. A R-evolution in power electronics: from "intelligent" to "super smart power" in automotive
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Marchio, F., primary, Poletto, V., additional, Russo, A., additional, Torrisi, G., additional, Notaro, J., additional, Burlak, G., additional, and Mirowski, M., additional
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29. VEGF-induced intracellular Ca2+ oscillations are down-regulated and do not stimulate angiogenesis in breast cancer-derived endothelial colony forming cells
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Valentina Poletto, Margherita Massa, Francesco Lodola, Dlzar Alì Khdar, Federico Alessandro Ruffinatti, Vittorio Rosti, Germano Guerra, Richard Tancredi, Alberto Riccardi, Marco Biggiogera, Estella Zuccolo, Francesco Moccia, Umberto Laforenza, Fabio Cattaneo, Lodola, F, Laforenza, U, Cattaneo, F, Ruffinatti, F, Poletto, V, Massa, M, Tancredi, R, Zuccolo, E, Khdar, D, Riccardi, A, Biggiogera, M, Rosti, V, Guerra, G, Moccia, F, Lodola, Francesco, Laforenza, Umberto, Cattaneo, Fabio, Ruffinatti Federico, Alessandro, Poletto, Valentina, Massa, Margherita, Tancredi, Richard, Zuccolo, Estella, Khdar Dlzar, Ali, Riccardi, Alberto, Biggiogera, Marco, Rosti, Vittorio, Guerra, Germano, and Moccia, Francesco.
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0301 basic medicine ,Angiogenesis ,VEGF receptors ,medicine.medical_treatment ,Population ,2+ ,Endothelial colony forming cells ,Intracellular Ca ,Breast cancer ,oscillations ,VEGF ,Endothelial colony forming cell ,Intracellular ca ,03 medical and health sciences ,0302 clinical medicine ,Medicine ,education ,education.field_of_study ,biology ,Intracellular Ca2+oscillations ,Oncology ,business.industry ,Angiogenesis, Breast cancer, Endothelial colony forming cells, Intracellular Ca2+ oscillations, VEGF ,Growth factor ,medicine.disease ,Molecular medicine ,Angiogenesi ,Intracellular Ca2+oscillation ,030104 developmental biology ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,business ,Intracellular - Abstract
// Francesco Lodola 1, 11, * , Umberto Laforenza 2, * , Fabio Cattaneo 3, * , Federico Alessandro Ruffinatti 4 , Valentina Poletto 5 , Margherita Massa 6 , Richard Tancredi 7 , Estella Zuccolo 1 , Dlzar Ali Khdar 1 , Alberto Riccardi 7, 8 , Marco Biggiogera 9 , Vittorio Rosti 5, ** , Germano Guerra 10, ** and Francesco Moccia 1 ** 1 Laboratory of General Physiology, Department of Biology and Biotechnology “Lazzaro Spallanzani”, University of Pavia, Pavia 27100, Italy 2 Department of Molecular Medicine, University of Pavia, Pavia 27100, Italy 3 Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples 80131, Italy 4 Department of Life Sciences and Systems Biology, Turin 10123, Italy 5 Laboratory of Biochemistry, Biotechnology and Advanced Diagnosis, Foundation IRCCS Policlinico San Matteo, Pavia 27100, Italy 6 Laboratory of Immunology Transplantation, Foundation IRCCS Policlinico San Matteo, Pavia 27100, Italy 7 Medical Oncology Unit, Foundation IRCCS Salvatore Maugeri, Pavia 27100, Italy 8 Department of Internal Medicine, University of Pavia, Pavia 27100, Italy 9 Laboratory of Cell Biology and Neurobiology, Department of Biology and Biotechnology “Lazzaro Spallanzani”, University of Pavia, Pavia 27100, Italy 10 Department of Medicine and Health Sciences “Vincenzo Tiberio”, University of Molise, Campobasso 86100, Italy 11 Current address: Italian Institute of Technology, Center for Nano Science and Technology, Milano 20133, Italy * These authors should be considered as co-first authors ** These authors share the Senior Authorship of the manuscript and should be regarded as co-last authors Correspondence to: Francesco Moccia, email: francesco.moccia@unipv.it Germano Guerra, email: germano.guerra@unipv.it Keywords: VEGF, breast cancer, endothelial colony forming cells, intracellular Ca 2+ oscillations, angiogenesis Received: June 01, 2017 Accepted: July 12, 2017 Published: August 14, 2017 ABSTRACT Endothelial colony forming cells (ECFCs) represent a population of truly endothelial precursors that promote the angiogenic switch in solid tumors, such as breast cancer (BC). The intracellular Ca 2+ toolkit, which drives the pro-angiogenic response to VEGF, is remodelled in tumor-associated ECFCs such that they are seemingly insensitive to this growth factor. This feature could underlie the relative failure of anti-VEGF therapies in cancer patients. Herein, we investigated whether and how VEGF uses Ca 2+ signalling to control angiogenesis in BC-derived ECFCs (BC-ECFCs). Although VEGFR-2 was normally expressed, VEGF failed to induce proliferation and in vitro tubulogenesis in BC-ECFCs. Likewise, VEGF did not trigger robust Ca 2+ oscillations in these cells. Similar to normal cells, VEGF-induced intracellular Ca 2+ oscillations were triggered by inositol-1,4,5-trisphosphate-dependent Ca 2+ release from the endoplasmic reticulum (ER) and maintained by store-operated Ca 2+ entry (SOCE). However, InsP 3 -dependent Ca 2+ release was significantly lower in BC-ECFCs due to the down-regulation of ER Ca 2+ levels, while there was no remarkable difference in the amplitude, pharmacological profile and molecular composition of SOCE. Thus, the attenuation of the pro-angiogenic Ca 2+ response to VEGF was seemingly due to the reduction in ER Ca 2+ concentration, which prevents VEGF from triggering robust intracellular Ca 2+ oscillations. However, the pharmacological inhibition of SOCE prevented BC-ECFC proliferation and in vitro tubulogenesis. These findings demonstrate for the first time that BC-ECFCs are insensitive to VEGF, which might explain at cellular and molecular levels the failure of anti-VEGF therapies in BC patients, and hint at SOCE as a novel molecular target for this disease.
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- 2017
30. Arachidonic acid-evoked Ca2+ signals promote nitric oxide release and proliferation in human endothelial colony forming cells
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Dmitry Lim, Alessandra Rappa, Francesco Lodola, Paolo Catarsi, Germano Guerra, Marta Reforgiato, Francesco Moccia, Estella Zuccolo, Silvia Dragoni, Vittorio Rosti, Daniele Guido, Valentina Poletto, Zuccolo, E, Dragoni, S, Poletto, V, Catarsi, P, Guido, D, Rappa, A, Reforgiato, M, Lodola, F, Lim, D, Rosti, V, Guerra, G, and Moccia, F
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Arachidonic acid, physiology, ECFCs, nitric oxide ,0301 basic medicine ,Physiology ,Proliferation ,Endothelial colony forming cells ,Arachidonic acid ,Ca(2+) ,Nitric oxide ,TRPV4 ,03 medical and health sciences ,chemistry.chemical_compound ,Transient receptor potential channel ,Enos ,Receptor ,Pharmacology ,biology ,Endoplasmic reticulum ,biology.organism_classification ,Cell biology ,Endothelial stem cell ,030104 developmental biology ,chemistry ,Biochemistry ,Molecular Medicine ,Intracellular - Abstract
Arachidonic acid (AA) stimulates endothelial cell (EC) proliferation through an increase in intracellular Ca2 + concentration ([Ca2 +]i), that, in turn, promotes nitric oxide (NO) release. AA-evoked Ca2 + signals are mainly mediated by Transient Receptor Potential Vanilloid 4 (TRPV4) channels. Circulating endothelial colony forming cells (ECFCs) represent the only established precursors of ECs. In the present study, we, therefore, sought to elucidate whether AA promotes human ECFC (hECFC) proliferation through an increase in [Ca2 +]i and the following activation of the endothelial NO synthase (eNOS). AA induced a dose-dependent [Ca2 +]i raise that was mimicked by its non-metabolizable analogue eicosatetraynoic acid. AA-evoked Ca2 + signals required both intracellular Ca2 + release and external Ca2 + inflow. AA-induced Ca2 + release was mediated by inositol-1,4,5-trisphosphate receptors from the endoplasmic reticulum and by two pore channel 1 from the acidic stores of the endolysosomal system. AA-evoked Ca2 + entry was, in turn, mediated by TRPV4, while it did not involve store-operated Ca2 + entry. Moreover, AA caused an increase in NO levels which was blocked by preventing the concomitant increase in [Ca2 +]i and by inhibiting eNOS activity with NG-nitro- l -arginine methyl ester ( l -NAME). Finally, AA per se did not stimulate hECFC growth, but potentiated growth factors-induced hECFC proliferation in a Ca2 +- and NO-dependent manner. Therefore, AA-evoked Ca2 + signals emerge as an additional target to prevent cancer vascularisation, which may be sustained by ECFC recruitment.
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- 2016
31. Stromal Cell-Derived Factor-1alpha Promotes Endothelial Colony-Forming Cell Migration Through the Ca2+-Dependent Activation of the Extracellular Signal-Regulated Kinase 1/2 and Phosphoinositide 3-Kinase/AKT Pathways
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Stefania Orecchioni, Dlzar A. Kheder, Vittorio Rosti, Christian A. Di Buduo, Valentina Poletto, Alessandra Balduini, Francesco Bertolini, Francesco Lodola, Francesco Moccia, Estella Zuccolo, Germano Guerra, Giorgia Scarpellino, Zuccolo, E, Di Buduo, C, Lodola, F, Orecchioni, S, Scarpellino, G, Kheder, D, Poletto, V, Guerra, G, Bertolini, F, Balduini, A, Rosti, V, and Moccia, F
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0301 basic medicine ,MAPK/ERK pathway ,Stromal cell ,Biology ,calcium signaling ,migration ,endothelial colony-forming cells ,03 medical and health sciences ,Extracellular ,Protein kinase B ,extracellular signal-regulated kinase ,PI3K/AKT/mTOR pathway ,Kinase ,Akt/PKB signaling pathway ,stromal cell-derived factor-1α ,Hematology ,Cell Biology ,extracellular signal-regulated kinases ,phosphoinositide 3-kinase/AKT ,stromal cell-derived factor-1α ,Cell biology ,030104 developmental biology ,endothelial colony-forming cell ,Intracellular ,Developmental Biology - Abstract
Stromal cell-derived factor-1α (SDF-1α) drives endothelial colony-forming cell (ECFC) homing and incorporation within neovessels, thereby restoring tissue perfusion in ischemic tissues and favoring tumor vascularization and metastasis. SDF-1α stimulates ECFC migration by activating the Gi-protein-coupled receptor, CXCR4, and then engaging the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway. Sporadic evidence showed that SDF-1α may also act through an increase in intracellular Ca2+ concentration ([Ca2+]i) in bone marrow-derived hematopoietic progenitor cells and fully differentiated endothelial cells. Of note, recent evidence demonstrated that intracellular Ca2+ signals play a key role in controlling the proangiogenic activity of ECFCs. The present investigation was, therefore, undertaken to assess whether and how SDF-1α induces ECFC motility by triggering intracellular Ca2+ signals. We found that SDF-1α caused a dose-dependent increase in [Ca2+]i that was inhibited by ADM3100, a selective CXCR4 antagonist. Pharmacological manipulation revealed that the Ca2+ response to [Ca2+]i was shaped by an initial intracellular Ca2+ release through inositol-1,4,5-trisphosphate receptors (InsP3Rs), followed by a sustained phase of extracellular Ca2+ entry through store-operated Ca2+ channels. InsP3-dependent Ca2+ release and store-operated Ca2+ entry (SOCE) were both necessary for SDF-1α-induced extracellular signal-regulated kinases 1/2 (ERK 1/2) and AKT phosphorylation. Finally, SDF-1α employed intracellular Ca2+ signals, ERK 1/2, and PI3K/AKT to promote ECFC migration in vitro and neovessel formation in vivo. These data, therefore, provide the first evidence that SDF-1α induces ECFC migration through the Ca2+-dependent activation of the ERK 1/2 and PI3K/AKT pathways.
- Published
- 2018
32. VEGF-induced intracellular Ca2+ oscillations are weaker and do not stimulate proliferation in tumor-derived endothelial colony forming cells
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Germano Guerra, Francesco Lodola, Umberto Laforenza, Fabio Cattaneo, Valentina Poletto, Estella Zuccolo, Marco Biggiogera, Vittorio Rosti, Domenico Tafuri, Francesco Moccia, Guerra, G, Lodola, F, Laforenza, U, Cattaneo, F, Poletto, V, Zuccolo, E, Biggiogera, M, Rosti, V, Tafuri, D, and Moccia, F
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VEGF ,breast cancer ,endothelial colony forming cells ,intracellular Ca2+ oscillations ,proliferation ,Physiology - Abstract
Endothelial colony forming cells (ECFCs) represent a population of truly endothelial precursors that may be mobilized from their vascular stem cell niches to promote the angiogenic switch in a growing number of solid malignancies, including breast cancer (BC). While normal ECFCs require VEGF to proliferate, tumor-associated ECFCs are seemingly insensitive to this growth factor. This phenomenon could contribute to the relative failure of anti-VEGF therapies in cancer patients. Recent work showed that the intracellular Ca2+ toolkit, which is a crucial determinant of ECFC fate and controls the pro-angiogenic program triggered by VEGF, is remodelled in tumor-associated ECFCs. Herein, we adopted an array of techniques, including Ca2+ imaging, electron microscopy, flow cytometry, real-time polymerase chain reaction, western blot analysis and functional assay to investigate whether and how VEGF uses Ca2+ signalling to control proliferation in BC-derived ECFCs (BC-ECFCs). Our results finally demonstrate for the first time that BC-ECFCs are insensitive to VEGF, which might explain at cellular and molecular level the failure of anti-VEGF therapies in BC patients, and hint at SOCE as a novel molecular target for this disease., Italian Journal of Anatomy and Embryology, Vol. 122, No. 1 (Supplement) 2017
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- 2017
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33. Liposomes as a Putative Tool to Investigate NAADP Signaling in Vasculogenesis
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Antonio De Luca, Vittorio Rosti, Valentina Poletto, Francesca Di Nezza, Estella Zuccolo, Luigi Ambrosone, Francesco Moccia, Germano Guerra, Di Nezza, F., Zuccolo, E., Poletto, V., Rosti, V., De Luca, A., Moccia, F., Guerra, G., and Ambrosone, L.
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0301 basic medicine ,Adult ,Male ,Ca ,Angiogenesis ,NAADP ,2+ ,Neovascularization, Physiologic ,LIPOSOMES ,Biology ,Biochemistry ,Piperazines ,Cell membrane ,03 medical and health sciences ,chemistry.chemical_compound ,Vasculogenesis ,BAPTA ,medicine ,Humans ,Calcium Signaling ,Molecular Biology ,Egtazic Acid ,ENDOTHELIAL PROGENITOR CELLS ,Cell Proliferation ,Endothelial Cell ,Nicotinic acid adenine dinucleotide phosphate ,Ca2+ SIGNALING ,VASCULOGENESIS ,Cell growth ,Endothelial Cells ,Cell Biology ,Cell biology ,Liposome ,030104 developmental biology ,medicine.anatomical_structure ,Carboline ,chemistry ,SIGNALING ,Second messenger system ,Female ,Intracellular ,NADP ,Carbolines ,Human - Abstract
Nicotinic acid adenine dinucleotide phosphate (NAADP) is the newest discovered intracellular second messengers, which is able to release Ca2+ stored within endolysosomal (EL) vesicles. NAADP-induced Ca2+ signals mediate a growing number of cellular functions, ranging from proliferation to muscle contraction and differentiation. Recently, NAADP has recently been shown to regulate angiogenesis by promoting endothelial cell growth. It is, however, still unknown whether NAADP stimulates proliferation also in endothelial progenitor cells, which are mobilized in circulation after an ischemic insult to induce tissue revascularization. Herein, we described a novel approach to prepare NAADP-containing liposomes, which are highly cell membrane permeable and are therefore amenable for stimulating cell activity. Accordingly, NAADP-containing liposomes evoked an increase in intracellular Ca2+ concentration, which was inhibited by NED-19, a selective inhibitor of NAADP-induced Ca2+ release. Furthermore, NAADP-containing liposomes promoted EPC proliferation, a process which was inhibited by NED-19 and BAPTA, a membrane permeable intracellular Ca2+ buffer. Therefore, NAADP-containing liposomes stand out as a promising tool to promote revascularization of hypoxic/ischemic tissues by favoring EPC proliferation. J. Cell. Biochem. 118: 3722-3729, 2017. © 2017 Wiley Periodicals, Inc.
- Published
- 2017
34. A3669G polymorphism of glucocorticoid receptor is a susceptibility allele for primary myelofibrosis and contributes to phenotypic diversity and blast transformation
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Vittorio Rosti, Laura Villani, Valentina Poletto, M. Martinetti, Anna Rita Migliaccio, Gianluca Viarengo, Paolo Catarsi, Alberto Malovini, Giovanni Barosi, Adriana Carolei, Rita Campanelli, Margherita Massa, Poletto, V, Rosti, V, Villani, L, Catarsi, P, Carolei, A, Campanelli, R, Massa, M, Martinetti, M, Viarengo, G, Malovini, A, FRANCO MIGLIACCIO, ANNA RITA, and Barosi, G
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Adult ,Male ,Adolescent ,Immunology ,Single-nucleotide polymorphism ,Biology ,Lymphocyte Activation ,Polymorphism, Single Nucleotide ,Biochemistry ,Cohort Studies ,Young Adult ,Receptors, Glucocorticoid ,Polycythemia vera ,Glucocorticoid receptor ,Polymorphism (computer science) ,White blood cell ,Genotype ,medicine ,Humans ,Genetic Predisposition to Disease ,Allele ,Child ,A3669G polymorphism glucocorticoid receptor primary myelofibrosis ,Myelofibrosis ,Aged ,Aged, 80 and over ,Myeloid Neoplasia ,Cell Biology ,Hematology ,Janus Kinase 2 ,Middle Aged ,Prognosis ,medicine.disease ,Survival Rate ,Phenotype ,medicine.anatomical_structure ,Primary Myelofibrosis ,Case-Control Studies ,Mutation ,Female ,Follow-Up Studies - Abstract
The frequency of A3669G single nucleotide polymorphism (SNP) of human glucocorticoid receptor has been reported increased in polycythemia vera. We investigated the frequency of A3669G SNP and its impact on disease phenotype and progression in 499 patients with primary myelofibrosis (PMF). The distribution of the A3669G allele differed between PMF patients and 2 healthy control populations (odds ratio, 1.6 and 1.8). The variant allele at the homozygous state (G/G) was associated with higher white blood cell count, larger spleen index, and higher frequency of circulating CD34+ cells at diagnosis. The latter association remained significant after correction for the JAK2V617F genotype. In patients JAK2V617F mutated, the G/G genotype was associated with shorter overall survival (77.6 months vs 298 months, P = .049) and blast transformation (BT)–free survival (76.7 months vs 261 months; P = .018). The latter association remained significant after correction for the known BT risk factors, such as age, sex, white blood cell count, percentage of blasts, IPSS prognostic score, and homozygosity for JAK2V617F (hazard ratio = 3.3; P = .006). In conclusion, the glucocorticoid receptor A3669G is a susceptibility allele for PMF: it contributes to confer the phenotype of excess myeloproliferation, and it cooperates with the JAK2V617F mutation in determining BT.
- Published
- 2012
35. Endoplasmic Reticulum Ca(2+) Handling and Apoptotic Resistance in Tumor-Derived Endothelial Colony Forming Cells
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Poletto, Valentina, Dragoni, Silvia, Lim, Dmitry, Biggiogera, Marco, Aronica, Adele, Cinelli, Mariapia, De Luca, Antonio, Rosti, Vittorio, Porta, Camillo, Guerra, Germano, Moccia, Francesco, Poletto, V, Dragoni, S, Lim, D, Biggiogera, M, Aronica, A, Cinelli, M, De Luca, A, Rosti, V, Porta, C, Guerra, G, and Moccia, Francesco
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CALCIUM SIGNALING ,APOPTOSIS ,CANCER ,ENDOTHELIAL PROGENITOR CELLS - Published
- 2016
36. A FUNCTIONAL TRANSIENT RECEPTOR POTENTIAL VANILLOID 4 (TRPV4) CHANNEL IS EPXRESSED IN HUMAN ENDOTHELIAL PROGENITOR CELLS
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Silvia, Dragoni, Germano, Guerra, Alessandra, Fiorio Pla, Giuseppe, Bertoni, Alessandra, Rappa, Valentina, Poletto, Cinzia, Bottino, Adele, Aronica, Francesco, Lodola, Maria Pia, Cinelli, Umberto, Laforenza, Vittorio, Rosti, Franco, Tanzi, Luca, Munaron, Francesco, Moccia, Dragoni, S, Guerra, G, Fiorio Pla, A, Bertoni, G, Rappa, A, Poletto, V, Bottino, C, Aronica, A, Lodola, F, Cinelli, M, Laforenza, U, Rosti, V, Tanzi, F, Munaron, L, and Moccia, F
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Adult ,Sulfonamides ,Stem Cells ,Endothelial Cells ,Neovascularization, Physiologic ,TRPV Cation Channels ,Ruthenium Red ,Young Adult ,Leucine ,Thiadiazoles ,Humans ,Tetradecanoylphorbol Acetate ,Anilides ,Calcium ,Endothelium, Vascular ,RNA, Messenger ,Ion channels, TRPV4, Physiology ,Cation Transport Proteins ,Cells, Cultured ,Cell Proliferation - Abstract
Endothelial progenitor cells (EPCs) are mobilized into circulation to replace damaged endothelial cells and recapitulate the vascular network of injured tissues. Intracellular Ca(2+) signals are key to EPC activation, but it is yet to be elucidated whether they are endowed with the same blend of Ca(2+) -permeable channels expressed by mature endothelial cells. For instance, endothelial colony forming cells (ECFCs), the only EPC subset truly committed to acquire a mature endothelial phenotype, lack canonical transient receptor potential channels 3, 5 and 6 (TRPC3, 5 and 6), which are widely distributed in vascular endothelium; on the other hand, they express a functional store-operated Ca(2+) entry (SOCE). The present study was undertaken to assess whether human circulating EPCs possess TRP vanilloid channel 4 (TRPV4), which plays a master signalling role in mature endothelium, by controlling both vascular remodelling and arterial pressure. We found that EPCs express both TRPV4 mRNA and protein. Moreover, both GSK1016790A (GSK) and phorbol myristate acetate and, two widely employed TRPV4 agonists, induced intracellular Ca(2+) signals uniquely in presence of extracellular Ca(2+). GSK- and PMA-induced Ca(2+) elevations were inhibited by RN-1734 and ruthenium red, which selectively target TRPV4 in mature endothelium. However, TRPV4 stimulation with GSK did not cause EPC proliferation, while the pharmacological blockade of TRPV4 only modestly affected EPC growth in the presence of a growth factor-enriched culture medium. Conversely, SOCE inhibition with BTP-2, La(3+) and Gd(3+) dramatically decreased cell proliferation. These data indicate that human circulating EPCs possess a functional TRPV4 protein before their engraftment into nascent vessels.
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- 2015
37. Dysregulation of VEGF-induced proangiogenic Ca2+ oscillations in primary myelofibrosis-derived endothelial colony-forming cells
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Valentina Poletto, Marta Reforgiato, Federico Alessandro Ruffinatti, Giovanni Barosi, Vittorio Rosti, Elisa Bonetti, Estella Zuccolo, Francesco Moccia, Silvia Dragoni, Germano Guerra, Francesco Lodola, Dragoni, S, Reforgiato, M, Zuccolo, E, Poletto, V, Lodola, F, Ruffinatti, F, Bonetti, E, Guerra, G, Barosi, G, Rosti, V, and Moccia, F
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Vascular Endothelial Growth Factor A ,medicine.medical_specialty ,Cancer Research ,Angiogenesis ,medicine.medical_treatment ,Cells ,Biology ,Endothelial progenitor cell ,Internal medicine ,medicine ,Genetics ,Cells, Cultured ,Endothelial Cells ,Female ,Humans ,Neovascularization, Pathologic ,Primary Myelofibrosis ,Stem Cells ,Calcium Signaling ,Hematology ,Molecular Biology ,Cell Biology ,human ,Progenitor cell ,phospholipase C ,Neovascularization ,Calcium signaling ,Pathologic ,calcium ,Cultured ,diacylglycerol ,inositol 1,4,5 trisphosphate ,vasculotropin ,vasculotropin A ,VEGFA protein, human ,Growth factor ,VEGFA protein ,Cell biology ,5 trisphosphate ,Vascular endothelial growth factor A ,Endocrinology ,Physiology, ECFCs, VEGF ,Stem cell ,Signal transduction ,inositol 1 - Abstract
Endothelial progenitor cells could be implicated in the aberrant neoangiogenesis that occurs in bone marrow and spleen in patients with primary myelofibrosis (PMF). However, antivascular endothelial growth factor (VEGF) monotherapy had only a modest and transient effect in these individuals. Recently it was found that VEGF-induced proangiogenic intracellular Ca 2+ oscillations could be impaired in endothelial progenitor cells of subjects with malignancies. Therefore, we employed Ca 2+ imaging, wavelet analysis, and functional assays to assess whether and how VEGF-induced Ca 2+ oscillations are altered in PMF-derived endothelial progenitor cells. We focused on endothelial colony-forming cells (ECFCs), which are the only endothelial progenitor cell subtype capable of forming neovessels both in vivo and in vitro. VEGF triggers repetitive Ca 2+ spikes in both normal ECFCs (N-ECFCs) and ECFCs obtained from PMF patients (PMF-ECFCs). However, the spiking response to VEGF is significantly weaker in PMF-ECFCs. VEGF-elicited Ca 2+ oscillations are patterned by the interaction between inositol-1,4,5-trisphosphate-dependent Ca 2+ mobilization and store-operated Ca 2+ entry. However, in most PMF-ECFCs, Ca 2+ oscillations are triggered by a store-independent Ca 2+ entry pathway. We found that diacylglycerol gates transient receptor potential canonical 1 channel to trigger VEGF-dependent Ca 2+ spikes by recruiting the phospholipase C/inositol-1,4,5-trisphosphate signaling pathway, reflected as a decrease in endoplasmic reticulum Ca 2+ content. Finally, we found that, apart from being less robust and dysregulated as compared with N-ECFCs, VEGF-induced Ca 2+ oscillations modestly stimulate PMF-ECFC growth and in vitro angiogenesis. These results may explain the modest effect of anti-VEGF therapies in PMF.
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- 2015
38. Enhanced expression of Stim, Orai, and TRPC transcripts and proteins in endothelial progenitor cells isolated from patients with primary myelofibrosis
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Umberto Laforenza, Silvia Dragoni, Adele Aronica, Franco Tanzi, Daniele Guido, Maria Rosa Guarrera, Valentina Poletto, Germano Guerra, Vittorio Rosti, Giovanni Barosi, Maria Pia Cinelli, Alessandra Rappa, Elisa Bonetti, Mario Tomasello, Marta Reforgiato, Francesco Lodola, Cinzia Bottino, Francesco Moccia, Sumedha Pareek, Dragoni, S, Laforenza, U, Bonetti, E, Reforgiato, M, Aronica, A, Lodola, F, Bottino, C, Guido, D, Rappa, A, Pareek, S, Tomasello, M, Guarrera, M, Cinelli, M, Poletto, V, Guerra, G, Barosi, G, Tanzi, F, Moccia, F, and Rosti, V
- Subjects
Male ,Physiology, ion channels ,Indoles ,Intracellular Space ,Gene Expression ,Gadolinium ,Cell Separation ,Inositol 1,4,5-Trisphosphate ,Endoplasmic Reticulum ,TRPC4 ,Membrane Potentials ,TRPC1 ,chemistry.chemical_compound ,Adenosine Triphosphate ,Molecular Cell Biology ,Signaling in Cellular Processes ,Anilides ,TRPC ,Endothelial Progenitor Cells ,Multidisciplinary ,ORAI1 ,Hematology ,STIM2 ,Middle Aged ,Signaling Cascades ,Cell biology ,Medicine ,Female ,Cellular Types ,Cyclopiazonic acid ,Signal Transduction ,Research Article ,Adult ,Science ,Biology ,Signaling Pathways ,Colony-Forming Units Assay ,Molecular Genetics ,Young Adult ,Lanthanum ,Thiadiazoles ,Genetics ,Calcium-Mediated Signal Transduction ,Humans ,RNA, Messenger ,Calcium Signaling ,Progenitor cell ,Aged ,Cell Proliferation ,TRPC Cation Channels ,Phospholipase C ,Membrane Proteins ,Computational Biology ,Endothelial Cells ,chemistry ,Primary Myelofibrosis ,Calcium Signaling Cascade ,Calcium ,Calcium Channels - Abstract
BackgroundAn increase in the frequency of circulating endothelial colony forming cells (ECFCs), the only subset of endothelial progenitor cells (EPCs) truly belonging to the endothelial phenotype, occurs in patients affected by primary myelofibrosis (PMF). Herein, they might contribute to the enhanced neovascularisation of fibrotic bone marrow and spleen. Store-operated Ca2+ entry (SOCE) activated by the depletion of the inositol-1,4,5-trisphosphate (InsP3)-sensitive Ca2+ store drives proliferation in ECFCs isolated from both healthy donors (N-ECFCs) and subjects suffering from renal cellular carcinoma (RCC-ECFCs). SOCE is up-regulated in RCC-ECFCs due to the over-expression of its underlying molecular components, namely Stim1, Orai1, and TRPC1.Methodology/principal findingsWe utilized Ca2+ imaging, real-time polymerase chain reaction, western blot analysis and functional assays to evaluate molecular structure and the functional role of SOCE in ECFCs derived from PMF patients (PMF-ECFCs). SOCE, induced by either pharmacological (i.e. cyclopiazonic acid or CPA) or physiological (i.e. ATP) stimulation, was significantly higher in PMF-ECFCs. ATP-induced SOCE was inhibited upon blockade of the phospholipase C/InsP3 signalling pathway with U73111 and 2-APB. The higher amplitude of SOCE was associated to the over-expression of the transcripts encoding for Stim2, Orai2-3, and TRPC1. Conversely, immunoblotting revealed that Stim2 levels remained constant as compared to N-ECFCs, while Stim1, Orai1, Orai3, TRPC1 and TRPC4 proteins were over-expressed in PMF-ECFCs. ATP-induced SOCE was inhibited by BTP-2 and low micromolar La3+ and Gd3+, while CPA-elicited SOCE was insensitive to Gd3+. Finally, BTP-2 and La3+ weakly blocked PMF-ECFC proliferation, while Gd3+ was ineffective.ConclusionsTwo distinct signalling pathways mediate SOCE in PMF-ECFCs; one is activated by passive store depletion and is Gd3+-resistant, while the other one is regulated by the InsP3-sensitive Ca2+ pool and is inhibited by Gd3+. Unlike N- and RCC-ECFCs, the InsP3-dependent SOCE does not drive PMF-ECFC proliferation.
- Published
- 2014
39. Functional and genetic aberrations of in vitro-cultured marrow-derived mesenchymal stromal cells of patients with classical Philadelphia-negative myeloproliferative neoplasms
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Daniela Ingo, Vittorio Rosti, G Barosi, Daniela Lisini, Francesco Locatelli, Laura Villani, Rita Maccario, Valentina Poletto, Valentina Achille, Elisa Bonetti, Marco Zecca, Gloria Acquafredda, Gabriela Fois, Rita Campanelli, Francesca Novara, Adele Aronica, M.A. Avanzini, Elisa Lenta, Paolo Catarsi, Maria Ester Bernardo, M. Massa, Orsetta Zuffardi, Robert Peter Gale, M. Mantelli, Antonia Moretta, Avanzini, M. A., Bernardo, M. E., Novara, F., Mantelli, M., Poletto, V., Villani, L., Lenta, E., Ingo, D. M., Achille, V., Bonetti, E., Massa, M., Campanelli, R., Fois, G., Catarsi, P., Gale, R. P., Moretta, A., Aronica, A., Maccario, R., Acquafredda, G., Lisini, D., Zecca, M., Zuffardi, O., Locatelli, F., Barosi, G., and Rosti, V.
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Bone Marrow Cells ,Philadelphia chromosome ,hemic and lymphatic diseases ,Medicine ,Humans ,Philadelphia Chromosome ,Cells, Cultured ,Philadelphia negative ,Chromosome Aberrations ,Myeloproliferative Disorders ,business.industry ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Hematology ,Gene deletion ,medicine.disease ,In vitro ,Oncology ,N/A ,Settore MED/38 - PEDIATRIA GENERALE E SPECIALISTICA ,business - Abstract
Functional and genetic aberrations of in vitro -cultured marrow-derived mesenchymal stromal cells of patients with classical Philadelphia-negative myeloproliferative neoplasms
- Published
- 2014
40. Combination of carotenoids from Spirulina and PLA/PLGA or PHB: New options to obtain bioactive nanoparticles.
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Santos Assunção L, Quênia Muniz Bezerra P, Stahl Hermes Poletto V, de Oliveira Rios A, Graça Ramos I, Duarte Ferreira Ribeiro C, Aparecida Souza Machado B, Izabel Druzian J, Alberto Vieira Costa J, and Larroza Nunes I
- Subjects
- Carotenoids isolation & purification, Drug Carriers chemistry, Excipients, Particle Size, Carotenoids pharmacology, Hydroxybutyrates chemistry, Nanoparticles chemistry, Polyesters chemistry, Polylactic Acid-Polyglycolic Acid Copolymer chemistry, Spirulina chemistry, beta Carotene chemistry
- Abstract
The use of poly-β-hydroxybutyrate (PHB) is an alternative polymer that can be considered environment friendly and renewable to prepare nanoparticles of carotenoids. This study aimed to develop and characterize aqueous dispersion nanoparticles and lyophilized nanoparticles of carotenoid extract obtained from Spirulina sp. LEB 18 by nanoprecipitation, using poly
d , l - lactic acid (PLA)/polyd , l - lactic-co-glycolic acid (PLGA) (75:25 w/w) or PHB as encapsulants. The samples were characterized for the particle size, polydispersity index, zeta potential, apparent viscosity, pH, color parameters, ultraviolet-visible (UV/Vis) spectrophotometry, carotenoid profile, encapsulation efficiency, morphology, and thermal analysis. Nanoparticles containing microalgae carotenoid extract showed average particle diameter on a nanoscale (<200 nm), high homogeneity and stability, high thermal stability, and encapsulation efficiency carotenoid (>80%) when compared to nanoparticles containing β-carotene synthetic. PHB or PLA/PLGA as encapsulating material in the production of nanoparticles from microalgae carotenoids can be a polymeric alternative capable of promoting greater stability and application of carotenoids., (Copyright © 2020 Elsevier Ltd. All rights reserved.)- Published
- 2021
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41. Kinetic and Angiogenic Activity of Circulating Endothelial Colony Forming Cells in Patients with Infantile Haemangioma Receiving Propranolol.
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Campanelli R, Codazzi AC, Poletto V, Abbà C, Catarsi P, Fois G, Avanzini MA, Brazzelli V, Tzialla C, De Silvestri A, Tinelli C, Licari A, Berra-Romani R, Zuccolo E, Moccia F, Mannarino S, Rosti V, and Massa M
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- Adrenergic beta-Antagonists therapeutic use, Antigens, CD34 metabolism, Calcium chemistry, Calcium Signaling, Cell Movement, Chemokine CXCL12 metabolism, Endothelial Cells drug effects, Endothelial Progenitor Cells cytology, Female, Flow Cytometry, Humans, Infant, Infant, Newborn, Kinetics, Leukocyte Common Antigens metabolism, Longitudinal Studies, Male, Mesenchymal Stem Cells cytology, Neovascularization, Physiologic, Phenotype, Receptors, CXCR4 metabolism, Endothelial Cells cytology, Hemangioma drug therapy, Hemangioma metabolism, Neovascularization, Pathologic, Propranolol therapeutic use
- Abstract
Endothelial progenitor cells (EPCs) have been suggested to contribute to the neovascularization of infantile haemangioma (IH). There is strong evidence of the efficacy of propranolol in the treatment of IH, possibly by inhibiting both vasculogenesis and angiogenesis in the tumour. We evaluate the frequency of circulating endothelial colony forming cells (ECFCs), as the best EPC surrogate, in patients with IH at diagnosis and while receiving propranolol by an ex vivo 12-month longitudinal study. Biological aspects of the ECFCs, such as their in vitro angiogenic potential, membrane CXCR4 expression and Ca
2+ signalling, were investigated. Circulating ECFCs were isolated by in vitro culture and expanded for 2 to 3 passages in 23 patients with IH (median age: 5.5 months, range: 5.5 weeks-11 months) before and 3, 6, 9 and 12 months after receiving propranolol. Twenty-four healthy subjects comparable for age were also assessed (CTRLs). Untreated patients with IH had a circulating ECFC frequency lower ( p = 0.001) than CTRLs; nevertheless, in in vitro starving conditions, ECFCs showed enhanced capacity to form tube-like structures than those of CTRLs. Patients with IH following the therapy with propranolol had a significantly increased ( p = 0.022) circulating ECFC frequency, that showed a diminished tube-like formation capacity in vitro, and an altered constitutive store-operated Ca2+ entry. ECFCs play a role in IH pathogenesis; the response to propranolol therapy is associated with their increased frequency in the peripheral blood and a reduction of their vasculogenic activity., Competing Interests: None declared., (Georg Thieme Verlag KG Stuttgart · New York.)- Published
- 2019
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42. The role of endothelial colony forming cells in kidney cancer's pathogenesis, and in resistance to anti-VEGFR agents and mTOR inhibitors: A speculative review.
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Poletto V, Rosti V, Biggiogera M, Guerra G, Moccia F, and Porta C
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- Endothelium, Vascular metabolism, Humans, Kidney Neoplasms drug therapy, Neovascularization, Pathologic drug therapy, Signal Transduction drug effects, Angiogenesis Inhibitors therapeutic use, Endothelium, Vascular pathology, Kidney Neoplasms blood supply, Kidney Neoplasms pathology, Neovascularization, Pathologic pathology, Receptors, Vascular Endothelial Growth Factor antagonists & inhibitors, TOR Serine-Threonine Kinases antagonists & inhibitors
- Abstract
Renal cell carcinoma (RCC) is highly dependent on angiogenesis, due to the overactivation of the VHL/HIF/VEGF/VEGFRs axis; this justifies the marked sensitivity of this neoplasm to antiangiogenic agents which, however, ultimately fail to control tumor growth. RCC also frequently shows alterations in the mTOR signaling pathway, and mTOR inhibitors have shown a similar pattern of initial activity/late failure as pure antiangiogenic agents. Understanding mechanisms of resistance to these agents would be key to improve the outcome of our patients. Circulating endothelial cells are a family of mainly bone marrow-derived progenitors, which have been postulated to be responsible of the reactivation of angiogenesis in different tumors. In this review, we shall discuss the complex nature and function of these cells, the evidence pro and contra their contribution to tumor vascularization, especially as far as RCC is concerned, and their possible role in determining resistance to presently available treatments., (Copyright © 2018. Published by Elsevier B.V.)
- Published
- 2018
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43. The spleen of patients with myelofibrosis harbors defective mesenchymal stromal cells.
- Author
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Avanzini MA, Abbonante V, Catarsi P, Dambruoso I, Mantelli M, Poletto V, Lenta E, Guglielmelli P, Croce S, Cobianchi L, Jemos B, Campanelli R, Bonetti E, Di Buduo CA, Salmoiraghi S, Villani L, Massa M, Boni M, Zappatore R, Iurlo A, Rambaldi A, Vannucchi AM, Bernasconi P, Balduini A, Barosi G, and Rosti V
- Subjects
- Adult, Aged, Aged, 80 and over, Antigens, CD34, Case-Control Studies, Cell Movement, Cell Proliferation, Female, Fibronectins metabolism, Hematopoiesis, Humans, Male, Matrix Metalloproteinase 2 metabolism, Megakaryocytes pathology, Middle Aged, Nestin metabolism, Young Adult, Mesenchymal Stem Cells pathology, Primary Myelofibrosis pathology, Spleen pathology
- Abstract
Splenic hematopoiesis is a major feature in the course of myelofibrosis (MF). In fact, the spleen of patients with MF contains malignant hematopoietic stem cells retaining a complete differentiation program, suggesting both a pivotal role of the spleen in maintaining the disease and a tight regulation of hematopoiesis by the splenic microenvironment, in particular by mesenchymal stromal cells (MSCs). Little is known about splenic MSCs (Sp-MSCs), both in normal and in pathological context. In this work, we have in vitro expanded and characterized Sp-MSCs from 25 patients with MF and 13 healthy subjects (HS). They shared similar phenotype, growth kinetics, and differentiation capacity. However, MF Sp-MSCs expressed significant lower levels of nestin, and favored megakaryocyte (Mk) differentiation in vitro at a larger extent than their normal counterpart. Moreover, they showed a significant upregulation of matrix metalloprotease 2 (MMP2) and fibronectin 1 (FN1) genes both at mRNA expression and at protein level, and, finally, developed genetic abnormalities which were never detected in HS-derived Sp-MSCs. Our data point toward the existence of a defective splenic niche in patients with MF that could be responsible of some pathological features of the disease, including the increased trafficking of CD34+ cells and the expansion of the megakaryocytic lineage., (© 2018 Wiley Periodicals, Inc.)
- Published
- 2018
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44. Stromal Cell-Derived Factor-1α Promotes Endothelial Colony-Forming Cell Migration Through the Ca 2+ -Dependent Activation of the Extracellular Signal-Regulated Kinase 1/2 and Phosphoinositide 3-Kinase/AKT Pathways.
- Author
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Zuccolo E, Di Buduo C, Lodola F, Orecchioni S, Scarpellino G, Kheder DA, Poletto V, Guerra G, Bertolini F, Balduini A, Rosti V, and Moccia F
- Subjects
- Adult, Animals, Humans, Inositol 1,4,5-Trisphosphate metabolism, Mice, Mitogen-Activated Protein Kinases metabolism, Receptors, CXCR4 metabolism, Signal Transduction physiology, Young Adult, Calcium metabolism, Cell Movement physiology, Chemokine CXCL12 metabolism, Endothelial Cells metabolism, MAP Kinase Signaling System physiology, Phosphatidylinositol 3-Kinases metabolism, Proto-Oncogene Proteins c-akt metabolism
- Abstract
Stromal cell-derived factor-1α (SDF-1α) drives endothelial colony-forming cell (ECFC) homing and incorporation within neovessels, thereby restoring tissue perfusion in ischemic tissues and favoring tumor vascularization and metastasis. SDF-1α stimulates ECFC migration by activating the G
i -protein-coupled receptor, CXCR4, and then engaging the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway. Sporadic evidence showed that SDF-1α may also act through an increase in intracellular Ca2+ concentration ([Ca2+ ]i ) in bone marrow-derived hematopoietic progenitor cells and fully differentiated endothelial cells. Of note, recent evidence demonstrated that intracellular Ca2+ signals play a key role in controlling the proangiogenic activity of ECFCs. The present investigation was, therefore, undertaken to assess whether and how SDF-1α induces ECFC motility by triggering intracellular Ca2+ signals. We found that SDF-1α caused a dose-dependent increase in [Ca2+ ]i that was inhibited by ADM3100, a selective CXCR4 antagonist. Pharmacological manipulation revealed that the Ca2+ response to [Ca2+ ]i was shaped by an initial intracellular Ca2+ release through inositol-1,4,5-trisphosphate receptors (InsP3 Rs), followed by a sustained phase of extracellular Ca2+ entry through store-operated Ca2+ channels. InsP3 -dependent Ca2+ release and store-operated Ca2+ entry (SOCE) were both necessary for SDF-1α-induced extracellular signal-regulated kinases 1/2 (ERK 1/2) and AKT phosphorylation. Finally, SDF-1α employed intracellular Ca2+ signals, ERK 1/2, and PI3K/AKT to promote ECFC migration in vitro and neovessel formation in vivo. These data, therefore, provide the first evidence that SDF-1α induces ECFC migration through the Ca2+ -dependent activation of the ERK 1/2 and PI3K/AKT pathways.- Published
- 2018
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45. Liposomes as a Putative Tool to Investigate NAADP Signaling in Vasculogenesis.
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Di Nezza F, Zuccolo E, Poletto V, Rosti V, De Luca A, Moccia F, Guerra G, and Ambrosone L
- Subjects
- Adult, Carbolines metabolism, Egtazic Acid analogs & derivatives, Egtazic Acid metabolism, Endothelial Cells cytology, Female, Humans, Liposomes, Male, NADP pharmacology, Piperazines metabolism, Calcium Signaling drug effects, Cell Proliferation drug effects, Endothelial Cells metabolism, NADP analogs & derivatives, Neovascularization, Physiologic drug effects
- Abstract
Nicotinic acid adenine dinucleotide phosphate (NAADP) is the newest discovered intracellular second messengers, which is able to release Ca
2+ stored within endolysosomal (EL) vesicles. NAADP-induced Ca2+ signals mediate a growing number of cellular functions, ranging from proliferation to muscle contraction and differentiation. Recently, NAADP has recently been shown to regulate angiogenesis by promoting endothelial cell growth. It is, however, still unknown whether NAADP stimulates proliferation also in endothelial progenitor cells, which are mobilized in circulation after an ischemic insult to induce tissue revascularization. Herein, we described a novel approach to prepare NAADP-containing liposomes, which are highly cell membrane permeable and are therefore amenable for stimulating cell activity. Accordingly, NAADP-containing liposomes evoked an increase in intracellular Ca2+ concentration, which was inhibited by NED-19, a selective inhibitor of NAADP-induced Ca2+ release. Furthermore, NAADP-containing liposomes promoted EPC proliferation, a process which was inhibited by NED-19 and BAPTA, a membrane permeable intracellular Ca2+ buffer. Therefore, NAADP-containing liposomes stand out as a promising tool to promote revascularization of hypoxic/ischemic tissues by favoring EPC proliferation. J. Cell. Biochem. 118: 3722-3729, 2017. © 2017 Wiley Periodicals, Inc., (© 2017 Wiley Periodicals, Inc.)- Published
- 2017
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46. Primary myelofibrosis: Older age and high JAK2V617F allele burden are associated with elevated plasma high-sensitivity C-reactive protein levels and a phenotype of progressive disease.
- Author
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Barosi G, Massa M, Campanelli R, Fois G, Catarsi P, Viarengo G, Villani L, Poletto V, Bosoni T, Magrini U, Gale RP, and Rosti V
- Subjects
- Adolescent, Adult, Age Factors, Aged, Aged, 80 and over, Alleles, Female, Humans, Inflammation etiology, Male, Middle Aged, Mutation, Phenotype, Young Adult, C-Reactive Protein analysis, Disease Progression, Janus Kinase 2 genetics, Primary Myelofibrosis pathology
- Abstract
We measured plasma levels of high-sensitivity C-reactive protein (hs-CRP) in 526 subjects with primary myelofibrosis (PMF). Thirty-eight percent had an elevated hs-CRP level (≥0.3mg/dL). Elevated hs-CRP levels were associated with a progressive disease phenotype, including anemia, high white blood cell count, low platelet count, increased splenomegaly, increased risk of blast transformation, and worse survival. Age≥52years, but no other demographic characteristics, was associated with an elevated hs-CRP level in multivariable logistic regression (odds ratio [OR], 4.29; 95% CI, 2.73-6.77; P <0.001). Subjects with JAK2V617F mutation and an allele burden≥50% had an age-independent higher incidence of elevated hs-CRP level (OR=1.97; 95% CI,1.21-3.22; P=0.006) compared with a combined cohort of subjects with JAK2V617F <50% allele burden, CALR, MPL mutations, or no detectable driver mutations. Neither ASXL1 or EZH2 sub-clonal mutations, nor JAK2 46/1 haplotype or the A3669G polymorphism of glucocorticoid receptor were significantly associated with increased hs-CRP levels. Subjects with age≥52years and JAK2V617F with≥50% allele burden had a phenotype of progressive disease. Our data indicate that older age and high JAK2V617 allele burden are major determinants of inflammation in PMF, and are associated with disease progression., (Copyright © 2017 Elsevier Ltd. All rights reserved.)
- Published
- 2017
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47. VEGF-induced intracellular Ca 2+ oscillations are down-regulated and do not stimulate angiogenesis in breast cancer-derived endothelial colony forming cells.
- Author
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Lodola F, Laforenza U, Cattaneo F, Ruffinatti FA, Poletto V, Massa M, Tancredi R, Zuccolo E, Khdar DA, Riccardi A, Biggiogera M, Rosti V, Guerra G, and Moccia F
- Abstract
Endothelial colony forming cells (ECFCs) represent a population of truly endothelial precursors that promote the angiogenic switch in solid tumors, such as breast cancer (BC). The intracellular Ca
2+ toolkit, which drives the pro-angiogenic response to VEGF, is remodelled in tumor-associated ECFCs such that they are seemingly insensitive to this growth factor. This feature could underlie the relative failure of anti-VEGF therapies in cancer patients. Herein, we investigated whether and how VEGF uses Ca2+ signalling to control angiogenesis in BC-derived ECFCs (BC-ECFCs). Although VEGFR-2 was normally expressed, VEGF failed to induce proliferation and in vitro tubulogenesis in BC-ECFCs. Likewise, VEGF did not trigger robust Ca2+ oscillations in these cells. Similar to normal cells, VEGF-induced intracellular Ca2+ oscillations were triggered by inositol-1,4,5-trisphosphate-dependent Ca2+ release from the endoplasmic reticulum (ER) and maintained by store-operated Ca2+ entry (SOCE). However, InsP3 -dependent Ca2+ release was significantly lower in BC-ECFCs due to the down-regulation of ER Ca2+ levels, while there was no remarkable difference in the amplitude, pharmacological profile and molecular composition of SOCE. Thus, the attenuation of the pro-angiogenic Ca2+ response to VEGF was seemingly due to the reduction in ER Ca2+ concentration, which prevents VEGF from triggering robust intracellular Ca2+ oscillations. However, the pharmacological inhibition of SOCE prevented BC-ECFC proliferation and in vitro tubulogenesis. These findings demonstrate for the first time that BC-ECFCs are insensitive to VEGF, which might explain at cellular and molecular levels the failure of anti-VEGF therapies in BC patients, and hint at SOCE as a novel molecular target for this disease., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of interest.- Published
- 2017
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48. Breast and renal cancer-Derived endothelial colony forming cells share a common gene signature.
- Author
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Moccia F, Fotia V, Tancredi R, Della Porta MG, Rosti V, Bonetti E, Poletto V, Marchini S, Beltrame L, Gallizzi G, Da Prada GA, Pedrazzoli P, Riccardi A, Porta C, Zambelli A, and D'Incalci M
- Subjects
- Adult, Aged, Aged, 80 and over, Antibiotics, Antineoplastic pharmacology, Breast Neoplasms blood supply, Carcinoma, Renal Cell blood supply, Down-Regulation genetics, Endothelial Progenitor Cells physiology, Female, Gene Expression Regulation, Neoplastic, Genotype, Humans, Kidney Neoplasms blood supply, Male, Middle Aged, Neovascularization, Pathologic genetics, Phenotype, Sirolimus pharmacology, Transcriptome, Tumor Cells, Cultured, Up-Regulation genetics, Breast Carcinoma In Situ genetics, Breast Neoplasms genetics, Carcinoma, Ductal, Breast genetics, Carcinoma, Renal Cell genetics, Kidney Neoplasms genetics
- Abstract
Background: Neovascularisation supports the metastatic switch in many aggressive solid cancers. Tumour neovessels are mostly lined by endothelial cells sprouting from nearby capillaries, but they could also be contributed by circulating endothelial progenitor cells (EPCs). However, scant information is available about tumour-derived EPCs., Methods: We carried out the first thorough, unbiased comparison of phenotype, function and genotype of normal versus tumour-derived endothelial colony forming cells (ECFCs), a truly endothelial EPC subtype. We used healthy donors-derived ECFCs (N-ECFCs) as control for breast cancer (BC)- and renal cell carcinoma (RCC)-derived ECFCs., Results: We found that both BC- and RCC-ECFCs belong to the endothelial lineage. Normal and tumour-derived ECFCs did not differ in terms of proliferative and tubulogenic rates. However, RCC-ECFCs were more resistant to rapamycin-induced apoptosis, whereas BC-ECFCs were more sensitive as compared with N-ECFCs. Gene expression profiling revealed 382 differentially expressed genes (DEGs; 192 upregulated and 150 downregulated) and 71 DEGs (33 upregulated, 38 downregulated) when comparing, respectively, BC- and RCC-ECFCs with N-ECFCs. Nonetheless, BC- and RCC-derived ECFCs shared 35 DEGs, 10 of which were validated by qRT-PCR; such 35 DEGs are organised in a gene network centred on FOS., Conclusion: These results provide the first clear-cut evidence that BC- and RCC-derived ECFCs exhibit an altered gene expression profile as compared with N-ECFCs; yet, they share a common gene signature that could highlight novel and more specific targets to suppress tumour vascularisation., (Copyright © 2017 Elsevier Ltd. All rights reserved.)
- Published
- 2017
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49. Arachidonic acid-evoked Ca 2+ signals promote nitric oxide release and proliferation in human endothelial colony forming cells.
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Zuccolo E, Dragoni S, Poletto V, Catarsi P, Guido D, Rappa A, Reforgiato M, Lodola F, Lim D, Rosti V, Guerra G, and Moccia F
- Subjects
- Adult, Arachidonic Acid administration & dosage, Calcium Signaling drug effects, Cell Proliferation drug effects, Cells, Cultured, Dose-Response Relationship, Drug, Endoplasmic Reticulum metabolism, Humans, NG-Nitroarginine Methyl Ester pharmacology, Nitric Oxide Synthase Type III metabolism, Young Adult, Arachidonic Acid metabolism, Calcium metabolism, Endothelial Progenitor Cells metabolism, Nitric Oxide metabolism
- Abstract
Arachidonic acid (AA) stimulates endothelial cell (EC) proliferation through an increase in intracellular Ca
2+ concentration ([Ca2+ ]i ), that, in turn, promotes nitric oxide (NO) release. AA-evoked Ca2+ signals are mainly mediated by Transient Receptor Potential Vanilloid 4 (TRPV4) channels. Circulating endothelial colony forming cells (ECFCs) represent the only established precursors of ECs. In the present study, we, therefore, sought to elucidate whether AA promotes human ECFC (hECFC) proliferation through an increase in [Ca2+ ]i and the following activation of the endothelial NO synthase (eNOS). AA induced a dose-dependent [Ca2+ ]i raise that was mimicked by its non-metabolizable analogue eicosatetraynoic acid. AA-evoked Ca2+ signals required both intracellular Ca2+ release and external Ca2+ inflow. AA-induced Ca2+ release was mediated by inositol-1,4,5-trisphosphate receptors from the endoplasmic reticulum and by two pore channel 1 from the acidic stores of the endolysosomal system. AA-evoked Ca2+ entry was, in turn, mediated by TRPV4, while it did not involve store-operated Ca2+ entry. Moreover, AA caused an increase in NO levels which was blocked by preventing the concomitant increase in [Ca2+ ]i and by inhibiting eNOS activity with NG-nitro-l-arginine methyl ester (l-NAME). Finally, AA per se did not stimulate hECFC growth, but potentiated growth factors-induced hECFC proliferation in a Ca2+ - and NO-dependent manner. Therefore, AA-evoked Ca2+ signals emerge as an additional target to prevent cancer vascularisation, which may be sustained by ECFC recruitment., (Copyright © 2016 Elsevier Inc. All rights reserved.)- Published
- 2016
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50. Endoplasmic Reticulum Ca(2+) Handling and Apoptotic Resistance in Tumor-Derived Endothelial Colony Forming Cells.
- Author
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Poletto V, Dragoni S, Lim D, Biggiogera M, Aronica A, Cinelli M, De Luca A, Rosti V, Porta C, Guerra G, and Moccia F
- Subjects
- Adult, Carcinoma, Renal Cell metabolism, Case-Control Studies, Cell Proliferation, Cells, Cultured, Endoplasmic Reticulum metabolism, Endothelial Progenitor Cells metabolism, Female, Humans, Kidney Neoplasms metabolism, Male, Middle Aged, Mitochondria metabolism, Signal Transduction, Young Adult, Apoptosis, Calcium metabolism, Carcinoma, Renal Cell pathology, Endoplasmic Reticulum pathology, Endothelial Progenitor Cells pathology, Kidney Neoplasms pathology, Mitochondria pathology
- Abstract
Truly endothelial progenitor cells (EPCs) can be mobilized from bone marrow to support the vascular network of growing tumors, thereby sustaining the metastatic switch. Endothelial colony forming cells (ECFCs) are the only EPC subtype belonging to the endothelial phenotype and capable of incorporating within neovessels. The intracellular Ca(2+) machinery plays a key role in ECFC activation and is remodeled in renal cellular carcinoma-derived ECFCs (RCC-ECFCs). Particularly, RCC-ECFCs seems to undergo a drop in endoplasmic reticulum (ER) Ca(2+) concentration ([Ca(2+) ]ER ). This feature is remarkable when considering that inositol-1,4,5-trisphosphate (InsP3 )-dependent ER-to-mitochondria Ca(2+) transfer regulates the intrinsic apoptosis pathway. Herein, we sought to assess whether: (1) the [Ca(2+) ]ER and the InsP3 -induced ER-mitochondria Ca(2+) shuttle are reduced in RCC-ECFCs; and (2) the dysregulation of ER Ca(2+) handling leads to apoptosis resistance in tumor-derived cells. RCC-ECFCs displayed a reduction both in [Ca(2+) ]ER and in the InsP3 -dependent mitochondrial Ca(2+) uptake, while they expressed normal levels of Bcl-2 and Bak. The decrease in [Ca(2+) ]ER was associated to a remarkable ER expansion in RCC-ECFCs, which is a hallmark of ER stress, and did not depend on the remodeling of the Ca(2+) -transporting and the ER Ca(2+) -storing systems. As expected, RCC-ECFCs were less sensitive to rapamycin- and thapsigargin-induced apoptosis; however, buffering intracellular Ca(2+) levels with BAPTA dampened apoptosis in both cell types. Finally, store-operated Ca(2+) entry was seemingly uncoupled from the apoptotic machinery in RCC-ECFCs. Thus, the chronic underfilling of the ER Ca(2+) pool could confer a survival advantage to RCC-ECFCs and underpin RCC resistance to pharmacological treatment. J. Cell. Biochem. 117: 2260-2271, 2016. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)
- Published
- 2016
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