67 results on '"Romano, Mf"'
Search Results
2. A Survey on Consumers with Lactose Intolerance in Italy: Their Perception, Habits and Needs When Shopping for Groceries
- Author
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Maria Sole Facioni, Dominici, S, Farace, B, Apicella, A, Romano, Mf, and Tarabella, A
- Published
- 2022
3. Automatic Anonymization of Italian Legal Textual Documents using Deep Learning
- Author
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Licari, D, Romano, Mf, and Comande', G
- Subjects
Deep Learning ,pseudonymization ,Italian court documents ,NER ,anonymization, pseudonymization, GDPR, Italian court documents, NER, Deep Learning ,GDPR ,anonymization - Published
- 2022
4. Enhancement of cytosine arabinoside-induced apoptosis in human myeloblastic leukemia cells by NF-κB/Rel− specific decoy oligodeoxynucleotides
- Author
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Romano, MF, Lamberti, A, Bisogni, R, Tassone, P, Pagnini, D, Storti, G, Del Vecchio, L, Turco, MC, and Venuta, S
- Published
- 2000
- Full Text
- View/download PDF
5. Assessing the carcinogenic potential of low-dose exposures to chemical mixtures in the environment: the challenge ahead
- Author
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Goodson, WH, Lowe, L, Carpenter, DO, Gilbertson, M, Ali, AM, de Cerain Salsamendi, AL, Lasfar, A, Carnero, A, Azqueta, A, Amedei, A, Charles, AK, Collins, AR, Ward, A, Salzberg, AC, Colacci, A, Olsen, A-K, Berg, A, Barclay, BJ, Zhou, BP, Blanco-Aparicio, C, Baglole, CJ, Dong, C, Mondello, C, Hsu, C-W, Naus, CC, Yedjou, C, Curran, CS, Laird, DW, Koch, DC, Carlin, DJ, Felsher, DW, Roy, D, Brown, DG, Ratovitski, E, Ryan, EP, Corsini, E, Rojas, E, Moon, E-Y, Laconi, E, Marongiu, F, Al-Mulla, F, Chiaradonna, F, Darroudi, F, Martin, FL, Van Schooten, FJ, Goldberg, GS, Wagemaker, G, Nangami, G, Calaf, GM, Williams, G, Wolf, GT, Koppen, G, Brunborg, G, Lyerly, HK, Krishnan, H, Ab Hamid, H, Yasaei, H, Sone, H, Kondoh, H, Salem, HK, Hsu, H-Y, Park, HH, Koturbash, I, Miousse, IR, Scovassi, AI, Klaunig, JE, Vondracek, J, Raju, J, Roman, J, Wise, JP, Whitfield, JR, Woodrick, J, Christopher, JA, Ochieng, J, Fernando Martinez-Leal, J, Weisz, J, Kravchenko, J, Sun, J, Prudhomme, KR, Narayanan, KB, Cohen-Solal, KA, Moorwood, K, Gonzalez, L, Soucek, L, Jian, L, D'Abronzo, LS, Lin, L-T, Li, L, Gulliver, L, McCawley, LJ, Memeo, L, Vermeulen, L, Leyns, L, Zhang, L, Valverde, M, Khatami, M, Romano, MF, Chapellier, M, Williams, MA, Wade, M, Manjili, MH, Lleonart, M, Xia, M, Gonzalez, MJ, Karamouzis, MV, Kirsch-Volders, M, Vaccari, M, Kuemmerle, NB, Singh, N, Cruickshanks, N, Kleinstreuer, N, van Larebeke, N, Ahmed, N, Ogunkua, O, Krishnakumar, PK, Vadgama, P, Marignani, PA, Ghosh, PM, Ostrosky-Wegman, P, Thompson, P, Dent, P, Heneberg, P, Darbre, P, Leung, PS, Nangia-Makker, P, Cheng, QS, Robey, RB, Al-Temaimi, R, Roy, R, Andrade-Vieira, R, Sinha, RK, Mehta, R, Vento, R, Di Fiore, R, Ponce-Cusi, R, Dornetshuber-Fleiss, R, Nahta, R, Castellino, RC, Palorini, R, Abd Hamid, R, Langie, SAS, Eltom, S, Brooks, SA, Ryeom, S, Wise, SS, Bay, SN, Harris, SA, Papagerakis, S, Romano, S, Pavanello, S, Eriksson, S, Forte, S, Casey, SC, Luanpitpong, S, Lee, T-J, Otsuki, T, Chen, T, Massfelder, T, Sanderson, T, Guarnieri, T, Hultman, T, Dormoy, V, Odero-Marah, V, Sabbisetti, V, Maguer-Satta, V, Rathmell, WK, Engstrom, W, Decker, WK, Bisson, WH, Rojanasakul, Y, Luqmani, Y, Chen, Z, Hu, Z, Goodson, WH, Lowe, L, Carpenter, DO, Gilbertson, M, Ali, AM, de Cerain Salsamendi, AL, Lasfar, A, Carnero, A, Azqueta, A, Amedei, A, Charles, AK, Collins, AR, Ward, A, Salzberg, AC, Colacci, A, Olsen, A-K, Berg, A, Barclay, BJ, Zhou, BP, Blanco-Aparicio, C, Baglole, CJ, Dong, C, Mondello, C, Hsu, C-W, Naus, CC, Yedjou, C, Curran, CS, Laird, DW, Koch, DC, Carlin, DJ, Felsher, DW, Roy, D, Brown, DG, Ratovitski, E, Ryan, EP, Corsini, E, Rojas, E, Moon, E-Y, Laconi, E, Marongiu, F, Al-Mulla, F, Chiaradonna, F, Darroudi, F, Martin, FL, Van Schooten, FJ, Goldberg, GS, Wagemaker, G, Nangami, G, Calaf, GM, Williams, G, Wolf, GT, Koppen, G, Brunborg, G, Lyerly, HK, Krishnan, H, Ab Hamid, H, Yasaei, H, Sone, H, Kondoh, H, Salem, HK, Hsu, H-Y, Park, HH, Koturbash, I, Miousse, IR, Scovassi, AI, Klaunig, JE, Vondracek, J, Raju, J, Roman, J, Wise, JP, Whitfield, JR, Woodrick, J, Christopher, JA, Ochieng, J, Fernando Martinez-Leal, J, Weisz, J, Kravchenko, J, Sun, J, Prudhomme, KR, Narayanan, KB, Cohen-Solal, KA, Moorwood, K, Gonzalez, L, Soucek, L, Jian, L, D'Abronzo, LS, Lin, L-T, Li, L, Gulliver, L, McCawley, LJ, Memeo, L, Vermeulen, L, Leyns, L, Zhang, L, Valverde, M, Khatami, M, Romano, MF, Chapellier, M, Williams, MA, Wade, M, Manjili, MH, Lleonart, M, Xia, M, Gonzalez, MJ, Karamouzis, MV, Kirsch-Volders, M, Vaccari, M, Kuemmerle, NB, Singh, N, Cruickshanks, N, Kleinstreuer, N, van Larebeke, N, Ahmed, N, Ogunkua, O, Krishnakumar, PK, Vadgama, P, Marignani, PA, Ghosh, PM, Ostrosky-Wegman, P, Thompson, P, Dent, P, Heneberg, P, Darbre, P, Leung, PS, Nangia-Makker, P, Cheng, QS, Robey, RB, Al-Temaimi, R, Roy, R, Andrade-Vieira, R, Sinha, RK, Mehta, R, Vento, R, Di Fiore, R, Ponce-Cusi, R, Dornetshuber-Fleiss, R, Nahta, R, Castellino, RC, Palorini, R, Abd Hamid, R, Langie, SAS, Eltom, S, Brooks, SA, Ryeom, S, Wise, SS, Bay, SN, Harris, SA, Papagerakis, S, Romano, S, Pavanello, S, Eriksson, S, Forte, S, Casey, SC, Luanpitpong, S, Lee, T-J, Otsuki, T, Chen, T, Massfelder, T, Sanderson, T, Guarnieri, T, Hultman, T, Dormoy, V, Odero-Marah, V, Sabbisetti, V, Maguer-Satta, V, Rathmell, WK, Engstrom, W, Decker, WK, Bisson, WH, Rojanasakul, Y, Luqmani, Y, Chen, Z, and Hu, Z
- Abstract
Lifestyle factors are responsible for a considerable portion of cancer incidence worldwide, but credible estimates from the World Health Organization and the International Agency for Research on Cancer (IARC) suggest that the fraction of cancers attributable to toxic environmental exposures is between 7% and 19%. To explore the hypothesis that low-dose exposures to mixtures of chemicals in the environment may be combining to contribute to environmental carcinogenesis, we reviewed 11 hallmark phenotypes of cancer, multiple priority target sites for disruption in each area and prototypical chemical disruptors for all targets, this included dose-response characterizations, evidence of low-dose effects and cross-hallmark effects for all targets and chemicals. In total, 85 examples of chemicals were reviewed for actions on key pathways/mechanisms related to carcinogenesis. Only 15% (13/85) were found to have evidence of a dose-response threshold, whereas 59% (50/85) exerted low-dose effects. No dose-response information was found for the remaining 26% (22/85). Our analysis suggests that the cumulative effects of individual (non-carcinogenic) chemicals acting on different pathways, and a variety of related systems, organs, tissues and cells could plausibly conspire to produce carcinogenic synergies. Additional basic research on carcinogenesis and research focused on low-dose effects of chemical mixtures needs to be rigorously pursued before the merits of this hypothesis can be further advanced. However, the structure of the World Health Organization International Programme on Chemical Safety 'Mode of Action' framework should be revisited as it has inherent weaknesses that are not fully aligned with our current understanding of cancer biology.
- Published
- 2015
6. Definizione di criteri per la classificazione e la valutazione delle caratteristiche dell'offerta formativa universitaria
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ZAVARRONE, EMMA, Zavarrone, E, Zaccarin, S, Crocetta, C, Balbi, S, Romano, M, ZAVARRONE, EMMA, Romano MF, ZAVARRONE, EMMA, Zavarrone, E, Zaccarin, S, Crocetta, C, Balbi, S, Romano, M, ZAVARRONE, EMMA, and Romano MF
- Abstract
A oltre cinque anni dalla introduzione del nuovo ordinamento del sistema universitario e alla vigilia dell'attivazione delle nuove classi di laurea a partire dal 2008/09, appare importante condurre una riflessione sul primo periodo di realizzazione della riforma per disporre di elementi utili ad indirizzare al meglio le scelte future. Questo studio analizza la nuova offerta didattica privilegiando il punto di vista della comunicazione che gli Atenei hanno adottato per far conoscere - e apprezzare - gli obiettivi e i risultati previsti, anche rispetto alle potenzialità occupazionali, della propria offerta formativa al fine di porre i giovani e le loro famiglie di fronte ad una scelta consapevole in grado di valorizzare capacità e risorse di ciascuno. L'attenzione esplicita rivolta alla descrizione dei risultati dell'apprendimento non solo, come tradizionalmente avviene, in termini di conoscenze incluse nel curriculum ma, anche, in termini di abilità e competenze richieste per il rilascio di un determinato titolo ha costituito uno dei punti portanti della riforma del sistema universitario ribadita anche nelle Linee Guida emanate dal Mur per la definizione dei nuovi ordinamenti didattici secondo i principi e gli obiettivi lanciati dalla dichiarazione di Bologna nel 1999. La sfida a cui sono chiamati i sistemi universitari nel contesto europeo è quella di facilitare la comparazione dei titoli, favorendone il loro riconoscimento e incentivando la mobilità dei soggetti coinvolti. A questo fine, si sta diffondendo la pratica di descrivere i risultati attesi dell'apprendimento, comuni a tutti i laureati di un certo tipo, mediante i cosiddetti descrittori del titolo(qualification descriptor), sufficientemente generali per poter essere applicati ad una vasta gamma di discipline e profili e sufficientemente semplici per poter essere adottati dai diversi sistemi nazionali (il riferimento è alla serie dei ¿descrittori di Dublino¿, elaborata dai gruppi di esperti istituiti
- Published
- 2008
7. Severe aortic stenosis and myocardial function: diagnostic and prognostic usefulness of ultrasonic integrated backscatter analysis.
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Di Bello V, Giorgi D, Viacava P, Enrica T, Nardi C, Palagi C, Delle Donne MG, Verunelli F, Mariani MA, Grandjean J, Dell'Anna R, Di Cori A, Zucchelli G, Romano MF, Mariani M, Di Bello, Vitantonio, Giorgi, Davide, Viacava, Paolo, Enrica, Talini, and Nardi, Carmela
- Published
- 2004
8. Cell stemness, epithelial-to-mesenchymal transition, and immunoevasion: Intertwined aspects in cancer metastasis
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Paolo D'Arrigo, Vincenza Vigorito, Maria Fiammetta Romano, Simona Romano, Salvatore Russo, Martina Tufano, Romano, Simona, Tufano, M, D'Arrigo, P, Vigorito, V, Russo, S, and Romano, Mf
- Subjects
PD-L1 ,0301 basic medicine ,Cancer Research ,Epithelial-Mesenchymal Transition ,medicine.medical_treatment ,Cell ,B7-H1 Antigen ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Cancer immunotherapy ,Neoplasms ,Tumor Microenvironment ,medicine ,Humans ,Epithelial–mesenchymal transition ,Neoplasm Metastasis ,Cell Death ,biology ,Mesenchymal phenotype ,Mesenchymal stem cell ,Cancer ,medicine.disease ,Killer Cells, Natural ,030104 developmental biology ,medicine.anatomical_structure ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,Cancer stemne ,Cancer cell ,Disease Progression ,Neoplastic Stem Cells ,biology.protein ,Cancer research ,Immunoevasion ,Tumor Escape ,Disease Susceptibility - Abstract
Recent advances in tumor immunology, fostered by dramatic outcomes with cancer immunotherapy, have opened new scenarios in cancer metastasis. The cancer stemness/mesenchymal phenotype and an excess of immune suppressive signals are emerging as Intertwined aspects of human tumors. This review examines recent studies that explored the mechanistic links between cancer cell stemness and immunoevasion, and the evidence points to these key events in cancer metastasis as two sides of the same coin. This review also covers the mechanisms involved in tumor expression of programmed cell death ligand 1 (PD-L1), a major factor exploited by human neoplasias to suppress immune control. We highlight the convergence of mesenchymal traits and PD-L1 expression and examine the functions of this immune inhibitory molecule, which confers cancer cell resistance and aggressiveness.
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- 2020
9. Effects of glycoprotein IIb/IIIa antagonists: Anti platelet aggregation and beyond
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Roberta Rossini, Arturo Giordano, Maria Romano, Anna D'Angelillo, Giuseppe Musumeci, Stefano Messina, Enrico Coscioni, Giuseppe Biondi Zoccai, Simona Romano, Giordano, A, Musumeci, G, D'Angelillo, A, Rossini, R, Zoccai, Gb, Messina, S, Coscioni, E, Romano, S, and Romano, Mf.
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0301 basic medicine ,Blood Platelets ,congenital, hereditary, and neonatal diseases and abnormalities ,Platelet Aggregation ,Abciximab ,Clinical Biochemistry ,Eptifibatide ,Platelet Glycoprotein GPIIb-IIIa Complex ,030204 cardiovascular system & hematology ,Pharmacology ,Fibrinogen ,Antibodies ,03 medical and health sciences ,Immunoglobulin Fab Fragments ,0302 clinical medicine ,hemic and lymphatic diseases ,Monoclonal ,Medicine ,Animals ,Humans ,Platelet ,Randomized Controlled Trials as Topic ,business.industry ,Antibodies, Monoclonal ,hemic and immune systems ,Peptides ,Platelet Aggregation Inhibitors ,Tyrosine ,Tirofiban ,030104 developmental biology ,Immunology ,Platelet aggregation inhibitor ,business ,Glycoprotein IIb/IIIa ,circulatory and respiratory physiology ,medicine.drug - Abstract
Background: The use of inhibitors of glycoprotein IIb/IIIa (GPIIb/IIIa) has provided dramatic results in terms of the prevention of acute stent thrombosis and a reduction in major adverse coronary events in patients subjected to percutaneous coronary intervention. GPIIb/IIIa or αIIbβ3 is a member of the β3 subfamily of integrins, which also includes αVβ3. GPIIb/IIIa functions as a receptor for fibrinogen and several adhesion proteins sharing an arginine-glycine-aspartic acid (RGD) sequence. GPIIb/IIIa antagonists, through blockade of the receptor, prevent platelet aggregation. Among the three GPIIb/IIIa antagonists used in therapy, abciximab is an anti-β3 monoclonal antibo dy, while tirofiban and eptifibatide mimic the binding sequence of the fibrinogen ligand. Although antiplatelet aggregation represents the central function of GPIIb/IIIa inhibitors, further actions have been documented for these compounds. Objective: The aim of the present article is to review the structures and functions of GPIIb/IIIa antagonists and to highlight the clinical outcomes and results of randomized trials with these compounds. Hypotheses on the unexplored potential of GPIIb/IIIa antagonists will be put forward. Conclusion: GPIIb/IIIa inhibitors were developed to prevent platelet aggregation, however, these compounds can exert further biological functions, both platelet- and non-platelet-related. Large-scale studies comparing the efficacy and safety of GPIIb/IIIa antagonists are lacking. More insights into the functions of these compounds may lead to generation of novel small molecules able to antagonize platelet aggregation while promoting vascular repair.
- Published
- 2016
10. Identification of a highly suppressive Treg subset associated to immunotherapy response
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Martina Tufano, Simona Romano, Emilio Francesco Giunta, Maria Romano, D. Faicchia, Giuseppe Argenziano, Fortunato Ciardiello, N. Novizio, P. D'Arrigo, G. Matarese, Teresa Troiani, A. Rea, Vincenza Vigorito, C. Procaccini, V. De Falco, Giunta, Ef, Romano, Simona, D’Arrigo, P, Rea, A, Tufano, M, Matarese, G, Procaccini, C, Novizio, N, Vigorito, V, Faicchia, D, Argenziano, G, De Falco, V, Ciardiello, F, Romano, Mf, and Troiani, T
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Oncology ,business.industry ,medicine.medical_treatment ,Immunology ,medicine ,Identification (biology) ,Hematology ,Immunotherapy ,business - Published
- 2018
11. Overexpression of chromatin assembly factor-1 p60, poly(ADP-ribose) polymerase 1 and nestin predicts metastasizing behaviour of oral cancer
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Gennaro Ilardi, Alba Rocco, Angela Celetti, Stefania Staibano, Gaetano De Rosa, Massimo Mascolo, Maria Romano, Chiara Luise, Maria Luisa Vecchione, Simona Romano, Francesco Merolla, Maria Siano, Mascolo, Massimo, Ilardi, Gennaro, Romano, Mf, Celetti, A, Siano, M, Romano, S, Luise, C, Merolla, F, Rocco, Alba, Vecchione, Ml, DE ROSA, Gaetano, and Staibano, Stefania
- Subjects
Adult ,Male ,Histology ,PARP-1 ,Nerve Tissue Proteins ,Stem cell marker ,Pathology and Forensic Medicine ,Metastasis ,Nestin ,Intermediate Filament Proteins ,stem cells ,Predictive Value of Tests ,medicine ,Carcinoma ,Biomarkers, Tumor ,Humans ,Neoplasm Metastasis ,PARP inhibitors ,Aged ,CAF-1 ,Mouth neoplasm ,Aged, 80 and over ,biology ,CAF-1 p60 ,CD44 ,Cancer ,General Medicine ,Original Articles ,Middle Aged ,oral cancer ,medicine.disease ,Prognosis ,Molecular biology ,Up-Regulation ,Survival Rate ,stomatognathic diseases ,Chromatin Assembly Factor-1 ,biology.protein ,Carcinoma, Squamous Cell ,Disease Progression ,Immunohistochemistry ,Female ,Mouth Neoplasms ,Poly(ADP-ribose) Polymerases ,Follow-Up Studies ,Transcription Factors - Abstract
Mascolo M, Ilardi G, Romano M F, Celetti A, Siano M, Romano S, Luise C, Merolla F, Rocco A, Vecchione M L, De Rosa G & Staibano S (2012) Histopathology Overexpression of chromatin assembly factor-1 p60, poly(ADP-ribose) polymerase 1 and nestin predicts metastasizing behaviour of oral cancer Aims: The natural history of oral squamous cell carcinomas (OSCCs) is variable and difficult to predict. This study aimed to assess the value of the expression of poly(ADP-ribose) polymerase 1 (PARP-1), chromatin assembly factor-1 (CAF-1)/p60 and the stem cell markers CD133, CD166, CD44, CD44v6 and nestin as markers of outcome and progression-free survival in OSCC patients. Methods: Clinical data were collected from 66 patients (41 male and 25 female, aged 29–92 years) who underwent surgery for OSCC of the tongue, floor, lips, and palate. During follow-up (range: 12–131 months), 14 patients experienced relapse/metastasis and/or death. The study was performed by immunohistochemistry on paraffin-embedded tumour tissues, western blot analysis of tumour protein lysates and human cell lines, and RNA silencing assays. In addition, the human papillomavirus (HPV) status of primary tumours was evaluated by immunohistochemistry and viral subtyping. Univariate and multivariate analyses were performed to determine the correlation between these parameters and the clinical and pathological variables of the study population. Results and conclusions: We found that a PARP-1high/CAF-1 p60high/nestinhigh phenotype characterized the OSCCs with the worst prognosis (all HPV-negative). This may be of benefit in clinical management, since radio-enhancing anti-PARP-1 and/or anti-CAF-1/p60 agents may allow radioresistance to be bypassed in the nestin-overexpressing, metastasizing OSCC cells.
- Published
- 2012
12. Morphological and molecular features of apoptosis and its role in colorectal cancer prevention
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Sivero, L., Giuseppe Galloro, Formisano, C., Sollazzo, V., Quarto, G., Romano, S., Romano, M. F., Sivero, Luigi, Galloro, Giuseppe, Formisano, Cesare, Sollazzo, V, Quarto, Gennaro, Romano, S, and Romano, Mf
- Subjects
immunocompetent system ,colorectal cancer ,apoptosi - Abstract
Apoptosis is a form of cell suicide that is essential for the control of cell growth in multicellular organisms. Apoptosis has a very important biological significance in mammals, both during embryonic life and adulthood. Defects in the apoptosis machinery cause several human diseases, including cancer. The induction of apoptosis is now recognized as a major strategy for the treatment and prevention of cancer. Colorectal cancer is a leading cause of cancer death, with the highest prevalence in North America, Europe, Australia, and New Zealand. Aim of the present article is to review the mechanism and biological significance of apoptosis and highlight the efficacy of natural agents that target apoptosis in colo-rectal cancer prevention.
13. Tirofiban prevents the effects of SARS-CoV-2 spike protein on macrophage activation and endothelial cell death.
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Marrone L, Romano S, Albanese M, Giordano S, Morello A, Cimmino M, Di Giacomo V, Malasomma C, Romano MF, and Corcione N
- Abstract
SARS-CoV-2 viral-derived particles have been proposed to have a causal role in tissue inflammation. Macrophage is the culprit cell in the pathogenesis of destructive inflammatory response to the SARS-CoV-2 virus. We investigated whether the spike protein might play a role in perturbing the physiological process of resolution of inflammation. Using an in vitro model of M2 polarized macrophages, we found that recombinant spike protein produced typical M1 morphological features in these alternative differentiated cells. In the presence of spike, M2-macrophages lose their elongated morphology, become rounded and acquire a strong capability to stimulate lymphocyte activation and proliferation. Moreover, in M2 macrophages, spike activated the signal transducer and activator-1 (STAT1) the pivotal mediator of pro-inflammatory macrophages. We observed STAT1 activation also in endothelial cells cultured with recombinant spike, accompanied by Bax upregulation and cell death. Blockade of beta3 integrin with the RGD mimetic tirofiban reverted the spike-induced costimulatory effects on M2 macrophages. Also, tirofiban counteracted STAT1 and Bax activation in endothelial cells cultured with spike and reduced endothelial cell death. In conclusion, we found that some proinflammatory effects of the spike protein can involve the integrin pathway and provide elements supporting use of RGD mimetics against SARS-Cov-2., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)
- Published
- 2024
- Full Text
- View/download PDF
14. The FKBP51s Splice Isoform Predicts Unfavorable Prognosis in Patients with Glioblastoma.
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Giordano C, Marrone L, Romano S, Della Pepa GM, Donzelli CM, Tufano M, Capasso M, Lasorsa VA, Quintavalle C, Guerri G, Martucci M, Auricchio A, Gessi M, Sala E, Olivi A, Romano MF, and Gaudino S
- Subjects
- Humans, Prognosis, Female, Male, Middle Aged, Tumor-Associated Macrophages immunology, Tumor-Associated Macrophages metabolism, Aged, Biomarkers, Tumor metabolism, Biomarkers, Tumor genetics, Magnetic Resonance Imaging, Adult, Glioblastoma genetics, Glioblastoma pathology, Glioblastoma immunology, Glioblastoma metabolism, Glioblastoma mortality, Glioblastoma diagnostic imaging, Tacrolimus Binding Proteins genetics, Tacrolimus Binding Proteins metabolism, Tumor Microenvironment immunology, Protein Isoforms genetics, Protein Isoforms metabolism, Brain Neoplasms pathology, Brain Neoplasms genetics, Brain Neoplasms metabolism, Brain Neoplasms immunology, Brain Neoplasms mortality
- Abstract
The primary treatment for glioblastoma (GBM) is removing the tumor mass as defined by MRI. However, MRI has limited diagnostic and predictive value. Tumor-associated macrophages (TAM) are abundant in GBM tumor microenvironment (TME) and are found in peripheral blood (PB). FKBP51 expression, with its canonical and spliced isoforms, is constitutive in immune cells and aberrant in GBM. Spliced FKBP51s supports M2 polarization. To find an immunologic signature that combined with MRI could advance in diagnosis, we immunophenotyped the macrophages of TME and PB from 37 patients with GBM using FKBP51s and classical M1-M2 markers. We also determined the tumor levels of FKBP51s, PD-L1, and HLA-DR. Tumors expressing FKBP51s showed an increase in various M2 phenotypes and regulatory T cells in PB, indicating immunosuppression. Tumors expressing FKBP51s also activated STAT3 and were associated with reduced survival. Correlative studies with MRI and tumor/macrophages cocultures allowed to interpret TAMs. Tumor volume correlated with M1 infiltration of TME. Cocultures with spheroids produced M1 polarization, suggesting that M1 macrophages may infiltrate alongside cancer stem cells. Cocultures of adherent cells developed the M2 phenotype CD163/FKBP51s expressing pSTAT6, a transcription factor enabling migration and invasion. In patients with recurrences, increased counts of CD163/FKBP51s monocyte/macrophages in PB correlated with callosal infiltration and were accompanied by a concomitant decrease in TME-infiltrating M1 macrophages. PB PD-L1/FKBP51s connoted necrotic tumors. In conclusion, FKBP51s identifies a GBM subtype that significantly impairs the immune system. Moreover, FKBP51s marks PB macrophages associated with MRI features of glioma malignancy that can aid in patient monitoring., Significance: Our research suggests that by combining imaging with analysis of monocyte/macrophage subsets in patients with GBM, we can enhance our understanding of the disease and assist in its treatment. We discovered a similarity in the macrophage composition between the TME and PB, and through association with imaging, we could interpret macrophages. In addition, we identified a predictive biomarker that drew more attention to immune suppression of patients with GBM., (© 2024 The Authors; Published by the American Association for Cancer Research.)
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- 2024
- Full Text
- View/download PDF
15. Metabolic vulnerability of cancer stem cells and their niche.
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Marrone L, Romano S, Malasomma C, Di Giacomo V, Cerullo A, Abate R, Vecchione MA, Fratantonio D, and Romano MF
- Abstract
Cancer stem cells (CSC) are the leading cause of the failure of anti-tumor treatments. These aggressive cancer cells are preserved and sustained by adjacent cells forming a specialized microenvironment, termed niche, among which tumor-associated macrophages (TAMs) are critical players. The cycle of tricarboxylic acids, fatty acid oxidation path, and electron transport chain have been proven to play central roles in the development and maintenance of CSCs and TAMs. By improving their oxidative metabolism, cancer cells are able to extract more energy from nutrients, which allows them to survive in nutritionally defective environments. Because mitochondria are crucial bioenergetic hubs and sites of these metabolic pathways, major hopes are posed for drugs targeting mitochondria. A wide range of medications targeting mitochondria, electron transport chain complexes, or oxidative enzymes are currently investigated in phase 1 and phase 2 clinical trials against hard-to-treat tumors. This review article aims to highlight recent literature on the metabolic adaptations of CSCs and their supporting macrophages. A focus is provided on the resistance and dormancy behaviors that give CSCs a selection advantage and quiescence capacity in particularly hostile microenvironments and the role of TAMs in supporting these attitudes. The article also describes medicaments that have demonstrated a robust ability to disrupt core oxidative metabolism in preclinical cancer studies and are currently being tested in clinical trials., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Marrone, Romano, Malasomma, Di Giacomo, Cerullo, Abate, Vecchione, Fratantonio and Romano.)
- Published
- 2024
- Full Text
- View/download PDF
16. Adversarial Learning for MRI Reconstruction and Classification of Cognitively Impaired Individuals.
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Zhou X, Balachandra AR, Romano MF, Chin SP, Au R, and Kolachalama VB
- Abstract
Game theory-inspired deep learning using a generative adversarial network provides an environment to competitively interact and accomplish a goal. In the context of medical imaging, most work has focused on achieving single tasks such as improving image resolution, segmenting images, and correcting motion artifacts. We developed a dual-objective adversarial learning framework that simultaneously 1) reconstructs higher quality brain magnetic resonance images (MRIs) that 2) retain disease-specific imaging features critical for predicting progression from mild cognitive impairment (MCI) to Alzheimer's disease (AD). We obtained 3-Tesla, T1-weighted brain MRIs of participants from the Alzheimer's Disease Neuroimaging Initiative (ADNI, N=342) and the National Alzheimer's Coordinating Center (NACC, N = 190) datasets. We simulated MRIs with missing data by removing 50% of sagittal slices from the original scans (i.e., diced scans). The generator was trained to reconstruct brain MRIs using the diced scans as input. We introduced a classifier into the GAN architecture to discriminate between stable (i.e., sMCI) and progressive MCI (i.e., pMCI) based on the generated images to facilitate encoding of disease-related information during reconstruction. The framework was trained using ADNI data and externally validated on NACC data. In the NACC cohort, generated images had better image quality than the diced scans (Structural similarity (SSIM) index: 0.553 ± 0.116 versus 0.348 ± 0.108). Furthermore, a classifier utilizing the generated images distinguished pMCI from sMCI more accurately than with the diced scans (F1-score: 0.634 ± 0.019 versus 0.573 ± 0.028). Competitive deep learning has potential to facilitate disease-oriented image reconstruction in those at risk of developing Alzheimer's disease.
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- 2024
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17. Deep learning for risk-based stratification of cognitively impaired individuals.
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Romano MF, Zhou X, Balachandra AR, Jadick MF, Qiu S, Nijhawan DA, Joshi PS, Mohammad S, Lee PH, Smith MJ, Paul AB, Mian AZ, Small JE, Chin SP, Au R, and Kolachalama VB
- Abstract
Quantifying the risk of progression to Alzheimer's disease (AD) could help identify persons who could benefit from early interventions. We used data from the Alzheimer's Disease Neuroimaging Initiative (ADNI, n = 544, discovery cohort) and the National Alzheimer's Coordinating Center (NACC, n = 508, validation cohort), subdividing individuals with mild cognitive impairment (MCI) into risk groups based on cerebrospinal fluid amyloid-β levels and identifying differential gray matter patterns. We then created models that fused neural networks with survival analysis, trained using non-parcellated T1-weighted brain MRIs from ADNI data, to predict the trajectories of MCI to AD conversion within the NACC cohort (integrated Brier score: 0.192 [discovery], and 0.108 [validation]). Using modern interpretability techniques, we verified that regions important for model prediction are classically associated with AD. We confirmed AD diagnosis labels using postmortem data. We conclude that our framework provides a strategy for risk-based stratification of individuals with MCI and for identifying regions key for disease prognosis., Competing Interests: V.B.K. reports honoraria from invited scientific presentations not exceeding $5000/year. He also serves as a consultant to Davos Alzheimer’s Collaborative and AstraZeneca. R.A. is a scientific advisor to Signant Health and consultant to Biogen. The remaining authors declare no competing interests., (© 2023 The Author(s).)
- Published
- 2023
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18. Scaffold proteins of cancer signaling networks: The paradigm of FK506 binding protein 51 (FKBP51) supporting tumor intrinsic properties and immune escape.
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Marrone L, D'Agostino M, Giordano C, Giacomo VD, Urzini S, Malasomma C, Gammella MP, Tufano M, Romano S, and Romano MF
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- Humans, Signal Transduction, Tacrolimus Binding Proteins genetics, Tacrolimus Binding Proteins chemistry, Tacrolimus Binding Proteins metabolism, Neoplasms genetics
- Abstract
Scaffold proteins are crucial regulators of signaling networks, and their abnormal expression may favor the development of tumors. Among the scaffold proteins, immunophilin covers a unique role as 'protein-philin' (Greek 'philin' = friend) that interacts with proteins to guide their proper assembly. The growing list of human syndromes associated with the immunophilin defect underscores the biological relevance of these proteins that are largely opportunistically exploited by cancer cells to support and enable the tumor's intrinsic properties. Among the members of the immunophilin family, the FKBP5 gene was the only one identified to have a splicing variant. Cancer cells impose unique demands on the splicing machinery, thus acquiring a particular susceptibility to splicing inhibitors. This review article aims to overview the current knowledge of the FKBP5 gene functions in human cancer, illustrating how cancer cells exploit the scaffolding function of canonical FKBP51 to foster signaling networks that support their intrinsic tumor properties and the spliced FKBP51s to gain the capacity to evade the immune system., Competing Interests: The authors declare that they have no conflicts of interest to report regarding the present study., (© 2023 Marrone et al.)
- Published
- 2023
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19. FKBP51 plays an essential role in Akt ubiquitination that requires Hsp90 and PHLPP.
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Tufano M, Marrone L, D'Ambrosio C, Di Giacomo V, Urzini S, Xiao Y, Matuozzo M, Scaloni A, Romano MF, and Romano S
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- Humans, Phosphorylation, Signal Transduction, Ubiquitination, Melanoma genetics, Proto-Oncogene Proteins c-akt genetics, Proto-Oncogene Proteins c-akt metabolism, Tacrolimus Binding Proteins genetics, Tacrolimus Binding Proteins metabolism, HSP90 Heat-Shock Proteins genetics, HSP90 Heat-Shock Proteins metabolism, Phosphoprotein Phosphatases genetics, Phosphoprotein Phosphatases metabolism
- Abstract
FKBP51 plays a relevant role in sustaining cancer cells, particularly melanoma. This cochaperone participates in several signaling pathways. FKBP51 forms a complex with Akt and PHLPP, which is reported to dephosphorylate Akt. Given the recent discovery of a spliced FKBP51 isoform, in this paper, we interrogate the canonical and spliced isoforms in regulation of Akt activation. We show that the TPR domain of FKBP51 mediates Akt ubiquitination at K63, which is an essential step for Akt activation. The spliced FKBP51, lacking such domain, cannot link K63-Ub residues to Akt. Unexpectedly, PHLPP silencing does not foster phosphorylation of Akt, and its overexpression even induces phosphorylation of Akt. PHLPP stabilizes levels of E3-ubiquitin ligase TRAF6 and supports K63-ubiquitination of Akt. The interactome profile of FKBP51 from melanoma cells highlights a relevant role for PHLPP in improving oncogenic hallmarks, particularly, cell proliferation., (© 2023. The Author(s).)
- Published
- 2023
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20. Biological relevance of ZNF224 expression in chronic lymphocytic leukemia and its implication IN NF-kB pathway regulation.
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Catapano R, Sepe L, Toscano E, Paolella G, Chiurazzi F, Barbato SP, Bruzzese D, Arianna R, Grosso M, Romano S, Romano MF, Costanzo P, and Cesaro E
- Abstract
Chronic lymphocytic leukemia (CLL) is a heterogeneous disease, whose presentation and clinical course are highly variable. Identification of novel prognostic factors may contribute to improving the CLL classification and providing indications for treatment options. The zinc finger protein ZNF224 plays a key role in cell transformation, through the control of apoptotic and survival pathways. In this study, we evaluated the potential application of ZNF224 as a novel marker of CLL progression and therapy responsiveness. To this aim, we analyzed ZNF224 expression levels in B lymphocytes from CLL patients at different stages of the disease and in patients showing different treatment outcomes. The expression of ZNF224 was significantly increased in disease progression and dramatically decreased in patients in complete remission after chemotherapy. Gene expression correlation analysis performed on datasets of CLL patients revealed that ZNF224 expression was well correlated with that of some prognostic and predictive markers. Moreover, bioinformatic analysis coupled ZNF224 to NF-κB pathway, and experimental data demonstrated that RNA interference of ZNF224 reduced the activity of the NF-κB survival pathway in CLL cells. Consistently with a pro-survival role, ZNF224 knockdown raised spontaneous and drug-induced apoptosis and inhibited the proliferation of peripheral blood mononuclear cells from CLL patients. Our findings provide evidence for the involvement of ZNF224 in the survival of CLL cells via NF-κB pathway modulation, and also suggest ZNF224 as a prognostic and predictive molecular marker of CLL disease., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Catapano, Sepe, Toscano, Paolella, Chiurazzi, Barbato, Bruzzese, Arianna, Grosso, Romano, Romano, Costanzo and Cesaro.)
- Published
- 2022
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21. FKBP51 Affects TNF-Related Apoptosis Inducing Ligand Response in Melanoma.
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Tufano M, Cesaro E, Martinelli R, Pacelli R, Romano S, and Romano MF
- Abstract
Melanoma is one of the most immunogenic tumors and has the highest potential to elicit specific adaptive antitumor immune responses. Immune cells induce apoptosis of cancer cells either by soluble factors or by triggering cell-death pathways. Melanoma cells exploit multiple mechanisms to escape immune system tumoricidal control. FKBP51 is a relevant pro-oncogenic factor of melanoma cells supporting NF-κB-mediated resistance and cancer stemness/invasion epigenetic programs. Herein, we show that FKBP51-silencing increases TNF-related apoptosis-inducing ligand (TRAIL)-R2 (DR5) expression and sensitizes melanoma cells to TRAIL-induced apoptosis. Consistent with the general increase in histone deacetylases, as by the proteomic profile, the immune precipitation assay showed decreased acetyl-Yin Yang 1 (YY1) after FKBP51 depletion, suggesting an impaired repressor activity of this transcription factor. ChIP assay supported this hypothesis. Compared with non-silenced cells, a reduced acetyl-YY1 was found on the DR5 promoter, resulting in increased DR5 transcript levels. Using Crispr/Cas9 knockout (KO) melanoma cells, we confirmed the negative regulation of DR5 by FKBP51. We also show that KO cells displayed reduced levels of acetyl-EP300 responsible for YY1 acetylation, along with reduced acetyl-YY1. Reconstituting FKBP51 levels contrasted the effects of KO on DR5, acetyl-YY1, and acetyl-EP300 levels. In conclusion, our finding shows that FKBP51 reduces DR5 expression at the transcriptional level by promoting YY1 repressor activity. Our study supports the conclusion that targeting FKBP51 increases the expression level of DR5 and sensitivity to TRAIL-induced cell death, which can improve the tumoricidal action of immune cells., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Tufano, Cesaro, Martinelli, Pacelli, Romano and Romano.)
- Published
- 2021
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22. PD-L1 Expression Fluctuates Concurrently with Cyclin D in Glioblastoma Cells.
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Tufano M, D'Arrigo P, D'Agostino M, Giordano C, Marrone L, Cesaro E, Romano MF, and Romano S
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- Cell Line, Tumor, Cell Proliferation physiology, Fibroblasts metabolism, Flow Cytometry methods, Humans, B7-H1 Antigen metabolism, Brain Neoplasms metabolism, Cyclin D metabolism, Glioblastoma metabolism
- Abstract
Despite Glioblastoma (GBM) frequently expressing programmed cell death ligand-1 (PD-L1), treatment with anti-programmed cell death-1 (PD1) has not yielded brilliant results. Intratumor variability of PD-L1 can impact determination accuracy. A previous study on mouse embryonic fibroblasts (MEFs) reported a role for cyclin-D in control of PD-L1 expression. Because tumor-cell growth within a cancer is highly heterogeneous, we looked at whether PD-L1 and its cochaperone FKBP51s were influenced by cell proliferation, using U251 and SF767 GBM-cell-lines. PD-L1 was measured by Western blot, flow cytometry, confocal-microscopy, quantitative PCR (qPCR), CCND1 by qPCR, FKBP51s by Western blot and confocal-microscopy. Chromatin-Immunoprecipitation assay (xChIp) served to assess the DNA-binding of FKBP51 isoforms. In the course of cell culture, PD-L1 appeared to increase concomitantly to cyclin-D on G1/S transition, to decrease during exponential cell growth progressively. We calculated a correlation between CCND1 and PD-L1 gene expression levels. In the temporal window of PD-L1 and CCND1 peak, FKBP51s localized in ER. When cyclin-D declined, FKBP51s went nuclear. XChIp showed that FKBP51s binds CCND1 gene in a closed-chromatin configuration. Our finding suggests that the dynamism of PD-L1 expression in GBM follows cyclin-D fluctuation and raises the hypothesis that FKBP51s might participate in the events that govern cyclin-D oscillation.
- Published
- 2021
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23. Opposing roles for striatonigral and striatopallidal neurons in dorsolateral striatum in consolidating new instrumental actions.
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Smith ACW, Jonkman S, Difeliceantonio AG, O'Connor RM, Ghoshal S, Romano MF, Everitt BJ, and Kenny PJ
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- Animals, Behavior, Animal, Corpus Striatum cytology, Male, Mice, Mice, Inbred C57BL, Rats, Rats, Long-Evans, Receptors, Dopamine D1 genetics, Receptors, Dopamine D1 metabolism, Receptors, Dopamine D2 genetics, Receptors, Dopamine D2 metabolism, Conditioning, Operant, Corpus Striatum physiology, Neurons physiology
- Abstract
Comparatively little is known about how new instrumental actions are encoded in the brain. Using whole-brain c-Fos mapping, we show that neural activity is increased in the anterior dorsolateral striatum (aDLS) of mice that successfully learn a new lever-press response to earn food rewards. Post-learning chemogenetic inhibition of aDLS disrupts consolidation of the new instrumental response. Similarly, post-learning infusion of the protein synthesis inhibitor anisomycin into the aDLS disrupts consolidation of the new response. Activity of D1 receptor-expressing medium spiny neurons (D1-MSNs) increases and D2-MSNs activity decreases in the aDLS during consolidation. Chemogenetic inhibition of D1-MSNs in aDLS disrupts the consolidation process whereas D2-MSN inhibition strengthens consolidation but blocks the expression of previously learned habit-like responses. These findings suggest that D1-MSNs in the aDLS encode new instrumental actions whereas D2-MSNs oppose this new learning and instead promote expression of habitual actions., (© 2021. The Author(s).)
- Published
- 2021
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24. Combining Magnetic Resonance Imaging with Systemic Monocyte Evaluation for the Implementation of GBM Management.
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Giordano C, Sabatino G, Romano S, Della Pepa GM, Tufano M, D'Alessandris QG, Cottonaro S, Gessi M, Balducci M, Romano MF, Olivi A, Gaudino S, and Colosimo C
- Subjects
- Adult, Aged, Antigens, CD blood, Antigens, Differentiation, Myelomonocytic blood, B7-H1 Antigen blood, Female, Humans, Lipopolysaccharide Receptors blood, Male, Middle Aged, Prospective Studies, Receptors, Cell Surface blood, Tacrolimus Binding Proteins blood, Brain Neoplasms blood, Brain Neoplasms diagnostic imaging, Brain Neoplasms therapy, Flow Cytometry, Glioblastoma blood, Glioblastoma diagnostic imaging, Glioblastoma therapy, Magnetic Resonance Imaging, Monocytes metabolism, Real-Time Polymerase Chain Reaction
- Abstract
Magnetic resonance imaging (MRI) is the gold standard for glioblastoma (GBM) patient evaluation. Additional non-invasive diagnostic modalities are needed. GBM is heavily infiltrated with tumor-associated macrophages (TAMs) that can be found in peripheral blood. FKBP51s supports alternative-macrophage polarization. Herein, we assessed FKBP51s expression in circulating monocytes from 14 GBM patients. The M2 monocyte phenotype was investigated by qPCR and flow cytometry using antibodies against PD-L1, CD163, FKBP51s, and CD14. MRI assessed morphologic features of the tumors that were aligned to flow cytometry data. PD-L1 expression on circulating monocytes correlated with MRI tumor necrosis score. A wider expansion in circulating CD163/monocytes was measured. These monocytes resulted in a dramatic decrease in patients with an MRI diagnosis of complete but not partial surgical removal of the tumor. Importantly, in patients with residual tumor, most of the peripheral monocytes that in the preoperative stage were CD163/FKBP51s- had turned into CD163/FKBP51s+. After Stupp therapy, CD163/FKBP51s+ monocytes were almost absent in a case of pseudoprogression, while two patients with stable or true disease progression showed sustained levels in such circulating monocytes. Our work provides preliminary but meaningful and novel results that deserve to be confirmed in a larger patient cohort, in support of potential usefulness in GBM monitoring of CD163/FKBP51s/CD14 immunophenotype in adjunct to MRI.
- Published
- 2021
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25. Multimodal Microvascular Mapping for Head and Neck, Skull Base Research and Education: An Anatomical Donor Study.
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House AE, Romano MF, Orczykowski ME, Zumwalt A, and Devaiah AK
- Abstract
Objective This study was aimed to develop a method combining computed tomography (CT) and fluorescence imaging, allowing identification of microvasculature in anatomical donors and facilitating translational research and education. Methods We investigated homogeneity and radiopacity of 30 different mixtures including radiopaque substances povidone-iodine (Betadine), barium sulfate (BaSO
4 ), and bismuth subsalicylate (Pepto-Bismol) varying in suspension and dilution with agar, latex, or gelatin. Three candidate mixtures were selected for testing the extent of perfusion in renal vasculature to establish methodology. From these candidate mixtures, two were selected for mixture with fluorescein and infusion into cadavers based on their ability to perfuse renal vasculature. The extent to which these two candidate mixtures combined with fluorescein were able to perfuse vasculature in a cadaver head was used to determine which mixture was superior. Results BaSO4 and bismuth subsalicylate-based mixtures demonstrated superior opacity in vials. In terms of solidifying agents, gelatin-based mixtures demonstrated increased friability and lower melting points compared with the other agents, so only latex and agar-based mixtures were used moving forward past the vial stage. Combinations of BaSO4 and latex and BaSO4 and 3% agar were found to perfuse kidneys superiorly to the mixture containing bismuth subsalicylate. Finally, in cadaver heads, the mixture containing BaSO4 , agar, and fluorescein was found to perfuse the smallest vasculature. Conclusion A final combination of BaSO4 , 3% agar, and fluorescein proves to be a powerful and novel combination enabling CT imaging, fluorescence imaging, and dissection of vasculature. This paves the way for future translational research and education., Competing Interests: Conflict of Interest A.K.D. reports in addition, A.K.D. has a patent retractable endoscopic suction pending. All the other authors report no conflict of interest., (Thieme. All rights reserved.)- Published
- 2021
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26. Eradication of CSCs: the roadmap for curing cancer.
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Romano S, Cesaro E, Tufano M, and Romano MF
- Abstract
Competing Interests: CONFLICTS OF INTEREST The authors declare no potential conflicts of interest.
- Published
- 2020
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27. Precision Calcium Imaging of Dense Neural Populations via a Cell-Body-Targeted Calcium Indicator.
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Shemesh OA, Linghu C, Piatkevich KD, Goodwin D, Celiker OT, Gritton HJ, Romano MF, Gao R, Yu CJ, Tseng HA, Bensussen S, Narayan S, Yang CT, Freifeld L, Siciliano CA, Gupta I, Wang J, Pak N, Yoon YG, Ullmann JFP, Guner-Ataman B, Noamany H, Sheinkopf ZR, Park WM, Asano S, Keating AE, Trimmer JS, Reimer J, Tolias AS, Bear MF, Tye KM, Han X, Ahrens MB, and Boyden ES
- Subjects
- Animals, Artifacts, Brain metabolism, Brain pathology, Calcium-Binding Proteins, Cell Body metabolism, Green Fluorescent Proteins, Mice, Neurons metabolism, Neuropil, Zebrafish, Brain diagnostic imaging, Calcium metabolism, Cell Body pathology, Neurons pathology, Optical Imaging methods
- Abstract
Methods for one-photon fluorescent imaging of calcium dynamics can capture the activity of hundreds of neurons across large fields of view at a low equipment complexity and cost. In contrast to two-photon methods, however, one-photon methods suffer from higher levels of crosstalk from neuropil, resulting in a decreased signal-to-noise ratio and artifactual correlations of neural activity. We address this problem by engineering cell-body-targeted variants of the fluorescent calcium indicators GCaMP6f and GCaMP7f. We screened fusions of GCaMP to natural, as well as artificial, peptides and identified fusions that localized GCaMP to within 50 μm of the cell body of neurons in mice and larval zebrafish. One-photon imaging of soma-targeted GCaMP in dense neural circuits reported fewer artifactual spikes from neuropil, an increased signal-to-noise ratio, and decreased artifactual correlation across neurons. Thus, soma-targeting of fluorescent calcium indicators facilitates usage of simple, powerful, one-photon methods for imaging neural calcium dynamics., Competing Interests: Declaration of Interests O.A.S., C.L., K.D.P., W.M.P., and E.S.B. declare that they applied for a U.S. patent based on the work presented in this paper, Application No.: PCT/US2019/065773., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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28. Alternative macrophage polarisation associated with resistance to anti-PD1 blockade is possibly supported by the splicing of FKBP51 immunophilin in melanoma patients.
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Troiani T, Giunta EF, Tufano M, Vigorito V, Arrigo P, Argenziano G, Ciardiello F, Romano MF, and Romano S
- Subjects
- Aged, Antibodies, Monoclonal, Humanized therapeutic use, B7-H1 Antigen antagonists & inhibitors, Female, Humans, Immune Checkpoint Inhibitors therapeutic use, Immunotherapy methods, Macrophage Activation immunology, Macrophages metabolism, Male, Melanoma drug therapy, Melanoma metabolism, Middle Aged, Nivolumab therapeutic use, Protein Isoforms, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory metabolism, Drug Resistance, Neoplasm immunology, Macrophages immunology, Melanoma immunology, Tacrolimus Binding Proteins metabolism
- Abstract
Background: FKBP51 immunophilin is abundantly expressed by immune cells. Co-inhibitory immune receptor signalling generates the splicing isoform FKBP51s. Tregs stained by FKBP51s are increased in melanoma patients and their counts are associated with anti-CTLA-4 response. An expansion of FKBP51s
+ PD-L1+ monocytes was measured in a group of non-responding patients to anti-CTLA-4. The aim of this work was to confirm the predictive value of response of FKBP51s+ Tregs in a cohort of patients undergoing anti-PD1 treatment and shed light on a monocyte subset co-expressing PD-L1/FKBP51s., Methods: Co-cultures of organoids and autologous lymphocytes were used to confirm that tumour T-cell interaction can induce FKBP51s. PBMC immunophenotype and flow cytometry served to assess and monitor FKBP51s+ Treg and FKBP51s+ PD-L1+ monocytes in 22 advanced melanoma patients treated with anti-PD1. Silencing and overexpression of FKBP51s in human macrophages served to address the protein role in the tolerant macrophages' behaviour., Results: FKBP51s+ Tregs count was increased in responders and had a prognostic value. Non-responders showed an early increase in FKBP51s+ PD-L1+ monocytes during anti-PD1 treatment. Manipulation of FKBP51s modulated the macrophage-phenotype, with forced protein expression promoting aspects associated with tolerance., Conclusions: FKBP51s may guide in the selection and monitoring of melanoma patient candidates to immune-checkpoint-targeted therapy. Manipulation of FKBP51s may overcome resistance.- Published
- 2020
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29. Neural crest-derived tumor neuroblastoma and melanoma share 1p13.2 as susceptibility locus that shows a long-range interaction with the SLC16A1 gene.
- Author
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Avitabile M, Succoio M, Testori A, Cardinale A, Vaksman Z, Lasorsa VA, Cantalupo S, Esposito M, Cimmino F, Montella A, Formicola D, Koster J, Andreotti V, Ghiorzo P, Romano MF, Staibano S, Scalvenzi M, Ayala F, Hakonarson H, Corrias MV, Devoto M, Law MH, Iles MM, Brown K, Diskin S, Zambrano N, Iolascon A, and Capasso M
- Subjects
- Adrenal Gland Neoplasms pathology, Cell Differentiation genetics, Cell Movement genetics, Chromosomes, Human, Pair 1 genetics, Female, Genetic Predisposition to Disease, Genome-Wide Association Study, Humans, Male, Melanoma pathology, Neural Crest pathology, Neuroblastoma pathology, Polymorphism, Single Nucleotide genetics, Skin Neoplasms pathology, Melanoma, Cutaneous Malignant, Adrenal Gland Neoplasms genetics, Melanoma genetics, Monocarboxylic Acid Transporters genetics, Neuroblastoma genetics, Skin Neoplasms genetics, Symporters genetics
- Abstract
Neuroblastoma (NB) and malignant cutaneous melanoma (CMM) are neural crest cells (NCC)-derived tumors and may have a shared genetic basis, but this has not been investigated systematically by genome-wide association studies (GWAS). We took a three-staged approach to conduct cross-disease meta-analysis of GWAS for NB and CMM (2101 NB cases and 4202 controls; 12 874 CMM cases and 23 203 controls) to identify shared loci. Findings were replicated in 1403 NB cases and 1403 controls of European ancestry and in 636 NB, 508 CMM cases and 2066 controls of Italian origin. We found a cross-association at locus 1p13.2 (rs2153977, odds ratio = 0.91, P = 5.36 × 10-8). We also detected a suggestive (P < 10-7) NB-CMM cross-association at 2q37.1 with opposite effect on cancer risk. Pathway analysis of 110 NB-CMM risk loci with P < 10-4 demonstrated enrichment of biological processes such as cell migration, cell cycle, metabolism and immune response, which are essential of human NCC development, underlying both tumors. In vitro and in silico analyses indicated that the rs2153977-T protective allele, located in an NB and CMM enhancer, decreased expression of SLC16A1 via long-range loop formation and altered a T-box protein binding site. Upon depletion of SLC16A1, we observed a decrease of cellular proliferation and invasion in both NB and CMM cell lines, suggesting its role as oncogene. This is the largest study to date examining pleiotropy across two NC cell-derived tumors identifying 1p13.2 as common susceptibility locus for NB and CMM risk. We demonstrate that combining genome-wide association studies results across cancers with same origins can identify new loci common to neuroblastoma and melanoma arising from tissues which originate from neural crest cells. Our results also show 1p13.2 confer risk to neuroblastoma and melanoma by regulating SLC16A1., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2020
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30. Population imaging of neural activity in awake behaving mice.
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Piatkevich KD, Bensussen S, Tseng HA, Shroff SN, Lopez-Huerta VG, Park D, Jung EE, Shemesh OA, Straub C, Gritton HJ, Romano MF, Costa E, Sabatini BL, Fu Z, Boyden ES, and Han X
- Subjects
- Action Potentials physiology, Animals, Hippocampus diagnostic imaging, Mice, Optogenetics, Environmental Biomarkers genetics, Hippocampus cytology, Neurons physiology, Optical Imaging methods, Wakefulness physiology
- Abstract
A longstanding goal in neuroscience has been to image membrane voltage across a population of individual neurons in an awake, behaving mammal. Here we describe a genetically encoded fluorescent voltage indicator, SomArchon, which exhibits millisecond response times and is compatible with optogenetic control, and which increases the sensitivity, signal-to-noise ratio, and number of neurons observable several-fold over previously published fully genetically encoded reagents
1-8 . Under conventional one-photon microscopy, SomArchon enables the routine population analysis of around 13 neurons at once, in multiple brain regions (cortex, hippocampus, and striatum) of head-fixed, awake, behaving mice. Using SomArchon, we detected both positive and negative responses of striatal neurons during movement, as previously reported by electrophysiology but not easily detected using modern calcium imaging techniques9-11 , highlighting the power of voltage imaging to reveal bidirectional modulation. We also examined how spikes relate to the subthreshold theta oscillations of individual hippocampal neurons, with SomArchon showing that the spikes of individual neurons are more phase-locked to their own subthreshold theta oscillations than to local field potential theta oscillations. Thus, SomArchon reports both spikes and subthreshold voltage dynamics in awake, behaving mice.- Published
- 2019
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31. The splicing FK506-binding protein-51 isoform plays a role in glioblastoma resistance through programmed cell death ligand-1 expression regulation.
- Author
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D'Arrigo P, Digregorio M, Romano S, Tufano M, Rea A, Hausch F, Dedobbeleer M, Vigorito V, Russo S, Bauder M, Rogister B, and Romano MF
- Abstract
Gliomas aberrantly express programmed cell death ligand-1 (PD-L1), which has a pivotal role in immunoevasion. The splicing isoform of FKBP5 , termed FKBP51s, is a PD-L1 foldase, assisting the immune checkpoint molecule in maturation and expression on the plasma membrane. The concept that PD-L1 supports tumor-intrinsic properties is increasingly emerging. The aim of the present work was to confirm the pro-tumoral effect of PD-L1 on human glioma cell survival, stemness capacity and resistance, and to address the issue of whether, by targeting its foldase either chemically or by silencing, the aggressive tumor features could be attenuated. PD-L1-depleted glioma cells have a reduced threshold for apoptosis, while PD-L1 forced expression increases resistance. Similar results were obtained with FKBP51s modulation. The ability of PD-L1 to counteract cell death was hampered by FKBP51s silencing. PD-L1 expression was particularly high in glioma cells with a cancer-stem-cell profile. Moreover, PD-L1 sustained the spheroid formation capability of glioma cells. Targeting of FKBP51s by small-interfering RNA (siRNA) or the specific inhibitor SAFit2, reduced the number of formed spheroids, along with PD-L1 expression. Finally, in an orthotopic mouse model of glioblastoma, daily treatment with SAFit2 significantly reduced tumor PD-L1 expression, and tumor growth. In treated mice, caspase-3 activation and reduced vimentin expression were observed in excised tumors. In conclusion, targeting of FKBP51s hampers PD-L1 and its pro-tumoral properties, thereby affecting the self-renewal and growth capacities of glioblastoma cells in vitro and in vivo., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© The Author(s) 2019.)
- Published
- 2019
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32. Prognostic Value of a Tissue Doppler Index of Systodiastolic Function in Patients with Asymptomatic Heart Failure.
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Pugliese NR, Fabiani I, La Carrubba S, Carerj S, Conte L, Colonna P, Caso P, Benedetto F, Antonini-Canterin F, Romano MF, Citro R, and Di Bello V
- Abstract
Introduction: Doppler echocardiography with early diastolic transmitral velocity (E)/early mitral annular diastolic velocity (E') ratio has been proposed as the best predictor for evaluating left ventricle (LV) filling pressure. A dimensionless index E/(E' × S') ratio (S' = systolic mitral annulus velocity) resulted in readily, reproducible, and reliable predictor of LV filling pressure. We assessed the prognostic impact of E/E' × S') in patients with asymptomatic heart failure (HF)., Materials and Methods: We calculated E/(E' × S') in 337 patients (179 male, 53%; age 54.7 ± 13.7 years) using the average of septal and lateral mitral annular velocities. We considered a composite endpoint as follows: all-cause death, acute myocardial infarction, stroke, and HF exacerbation., Results: Baseline ejection fraction resulted 60.2 ± 11.8%; E/E' × S') was 1.45 ± 0.8, with S' 7.4 ± 2.4 cm/s and E/E' 9.5 ± 5.4. After a 22-month median follow-up, there were 42 events: 5 deaths (12%), 3 acute myocardial infarctions (7%), 1 stroke (2%), and 33 HF hospitalizations (79%). In patients reaching the composite endpoint, E/(E' × S') resulted 2.07 ± 1.1 versus 1.3 ± 0.7 in event-free population ( P < 0.001). In a Cox-regression analysis, adjusted for confounding clinical factors and conventional echo parameters, E/(E' × S') ( P < 0.001), age ( P < 0.001), and male gender ( P = 0.03) resulted independent predictors of the composite endpoint., Conclusions: E/(E' × S') was an independent predictor for the future cardiac events in asymptomatic HF., Competing Interests: There are no conflicts of interest.
- Published
- 2018
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33. FKBP51s signature in peripheral blood mononuclear cells of melanoma patients as a possible predictive factor for immunotherapy.
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Romano S, Simeone E, D'Angelillo A, D'Arrigo P, Russo M, Capasso M, Lasorsa VA, Zambrano N, Ascierto PA, and Romano MF
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- Aged, Female, Humans, Male, Immunophenotyping methods, Immunotherapy methods, Leukocytes, Mononuclear immunology, Melanoma immunology, Tacrolimus Binding Proteins metabolism
- Abstract
The inhibitory immune checkpoint PD-L1/PD1 promotes the alternative splicing of the FKBP5 gene, resulting in increased expression of its variant 4 in the peripheral blood mononuclear cells of melanoma patients. The variant 4 transcript is translated into the truncated FKBP51s protein. Given the importance of co-inhibitory signalling in tumour immune escape, here we tested the potential for using FKBP51s expression to predict immunotherapy outcomes. To do this, we immunophenotyped PBMCs from 118 melanoma patients and 77 age- and sex-matched healthy controls. Blood samples were collected before patients underwent ipilimumab treatment. In 64 of the 118 patients, FKBP51s expression was also assessed in regulatory T cells (Tregs). We found that each PBMC subset analysed contained an FKBP51s
pos fraction, and that this fraction was greater in the melanoma patients than healthy controls. In CD4 T lymphocytes, the FKBP51sneg fraction was significantly impaired. Tregs count was increased in melanoma patients, which is in line with previous studies. Also, by analyses of FKBP51s in Tregs, we identified a subgroup of ipilimumab nonresponder patients (p = 0.002). In conclusion, FKBP51s-based immunophenotyping of melanoma patients revealed several profiles related to a negative immune regulatory control and identified an unknown Treg subset. These findings are likely to be useful in the selection of the patients that are candidate for immunotherapy.- Published
- 2017
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34. A regulatory role for the co-chaperone FKBP51s in PD-L1 expression in glioma.
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D'Arrigo P, Russo M, Rea A, Tufano M, Guadagno E, Del Basso De Caro ML, Pacelli R, Hausch F, Staibano S, Ilardi G, Parisi S, Romano MF, and Romano S
- Abstract
Background: FKBP51 is a co-chaperone with isomerase activity, abundantly expressed in glioma. We previously identified a spliced isoform (FKBP51s) and highlighted a role for this protein in the upregulation of Programmed Death Ligand 1 (PD-L1) expression in melanoma. Because gliomas can express PD-L1 causing a defective host anti-tumoral immunity, we investigated whether FKBP51s was expressed in glioma and played a role in PD-L1 regulation in this tumour., Methods: We used D54 and U251 glioblastoma cell lines that constitutively expressed PD-L1. FKBP51s was measured by immunoblot, flow cytometry and microscopy. In patient tumours, IHC and qPCR were used to measure protein and mRNA levels respectively. FKBP51s depletion was achieved by siRNAs, and its enzymatic function was inhibited using selective inhibitors (SAFit). We investigated protein maturation using N-glycosidase and cell fractionation approaches., Results: FKBP51s was expressed at high levels in glioma cells. Glycosylated-PD-L1 was increased and reduced by FKBP51s overexpression or silencing, respectively. Naïve PD-L1 was found in the endoplasmic reticulum (ER) of glioma cells complexed with FKBP51s, whereas the glycosylated form was measured in the Golgi apparatus. SAFit reduced PD-L1 levels (constitutively expressed and ionizing radiation-induced). SAFit reduced cell death of PBMC co-cultured with glioma., Conclusions: Here we addressed the mechanism of post-translational regulation of PD-L1 protein in glioma. FKBP51s upregulated PD-L1 expression on the plasma membrane by catalysing the protein folding required for subsequent glycosylation. Inhibition of FKBP51s isomerase activity by SAFit decreased PD-L1 levels. These findings suggest that FKBP51s is a potential target of immunomodulatory strategies for glioblastoma treatment., Competing Interests: CONFLICTS OF INTEREST No conflicts of interest is declared.
- Published
- 2017
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35. FKBP51 Immunohistochemical Expression: A New Prognostic Biomarker for OSCC?
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Russo D, Merolla F, Mascolo M, Ilardi G, Romano S, Varricchio S, Napolitano V, Celetti A, Postiglione L, Di Lorenzo PP, Califano L, Dell'Aversana GO, Astarita F, Romano MF, and Staibano S
- Subjects
- Adult, Aged, Aged, 80 and over, Biomarkers, Tumor, Carcinoma, Squamous Cell diagnosis, Carcinoma, Squamous Cell therapy, Cyclin-Dependent Kinase Inhibitor p16 genetics, Cyclin-Dependent Kinase Inhibitor p16 metabolism, Female, Gene Expression, Human papillomavirus 16 physiology, Humans, Immunohistochemistry, Kaplan-Meier Estimate, Lymphatic Metastasis, Male, Middle Aged, Mouth Neoplasms diagnosis, Mouth Neoplasms therapy, Neoplasm Grading, Neoplasm Staging, Prognosis, Tacrolimus Binding Proteins genetics, Carcinoma, Squamous Cell metabolism, Carcinoma, Squamous Cell mortality, Mouth Neoplasms metabolism, Mouth Neoplasms mortality, Tacrolimus Binding Proteins metabolism
- Abstract
Up-to-date, several molecular markers of prognosis have been studied in Oral Squamous Cell Carcinoma (OSCC), but none entered in the clinical setting. Therapy of OSCC tumors mainly relies on surgery, radiotherapy and partially on chemotherapy; there is an urgent need for biomarkers able to better stratify OSCC patients' risk to address targeted therapeutic strategies. The role of immune response in the pathogenesis and biological behavior of OSCC has been investigated by several authors, and promising results have been obtained with immune checkpoint inhibitors. We already investigated the role of the immune modulator FK506-binding protein 51 (FKBP51), a FK506-binding immunophilin, in cutaneous melanoma biology, and its expression in several human solid tumors. In the present study, we aimed to assess the value of FKBP51 expression in OSCC tumor cells as a marker of outcome. We collected clinical data from 72 patients who underwent surgery for Squamous Cell Carcinoma (SCC) of the tongue, floor, lips and palate. FKBP51 expression was assessed by immunohistochemistry on paraffin-embedded tumor tissues. In addition, we evaluated the human papillomavirus (HPV) status of primary tumors by immunohistochemistry, viral subtyping and In Situ Hybridization (ISH) assay. We found that high FKBP51-expressing tumors characterized the OSCCs with the worst prognosis: the high immunohistochemical expression of FKBP51 associated with death occurring within five years from the diagnosis with a sensitivity of 88.46% and a specificity of 91.67%. The estimated positive predictive value of the test was 88.45% and negative predictive value 91.67%. We tested FKBP51 mRNA presence, by RT-PCR assay, in a selected series of OSCC tumors, and we found that mRNA correlated well to the protein expression and to the clinical outcome. Applying the Bayes formula, we estimated an 88% probability of dying within five years from the diagnosis of OSCC patients with a high FKBP51 immunohistochemical (IHC) test result (>51% of FKBP51 positive tumor cells). On the basis of our analysis, we propose tumor tissue expression of FKBP51 protein as a reliable prognostic marker for OSCC tumors.
- Published
- 2017
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36. Web Health Monitoring Survey: A New Approach to Enhance the Effectiveness of Telemedicine Systems.
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Romano MF, Sardella MV, and Alboni F
- Abstract
Background: Aging of the European population and interest in a healthy population in western countries have contributed to an increase in the number of health surveys, where the role of survey design, data collection, and data analysis methodology is clear and recognized by the whole scientific community. Survey methodology has had to couple with the challenges deriving from data collection through information and communications technology (ICT). Telemedicine systems have not used patients as a source of information, often limiting them to collecting only biometric data. A more effective telemonitoring system would be able to collect objective and subjective data (biometric parameters and symptoms reported by the patients themselves), and to control the quality of subjective data collected: this goal be achieved only by using and merging competencies from both survey methodology and health research., Objective: The objective of our study was to propose new metrics to control the quality of data, along with the well-known indicators of survey methodology. Web questionnaires administered daily to a group of patients for an extended length of time are a Web health monitoring survey (WHMS) in a telemedicine system., Methods: We calculated indicators based on paradata collected during a WHMS study involving 12 patients, who signed in to the website daily for 2 months., Results: The patients' involvement was very high: the patients' response rate ranged between 1.00 and 0.82, with an outlier of 0.65. Item nonresponse rate was very low, ranging between 0.0% and 7.4%. We propose adherence to the chosen time to connect to the website as a measure of involvement and cooperation by the patients: the difference from the median time ranged between 11 and 24 minutes, demonstrating very good cooperation and involvement from all patients. To measure habituation to the questionnaire, we also compared nonresponse rates to the items between the first and the second month of the study, and found no significant difference. We computed the time to complete the questionnaire both as a measure of possible burden for patient, and to detect the risk of automatic responses. Neither of these hypothesis was confirmed, and differences in time to completion seemed to depend on health conditions. Focus groups with patients confirmed their appreciation for this "new" active role in a telemonitoring system., Conclusions: The main and innovative aspect of our proposal is the use of a Web questionnaire to virtually recreate a checkup visit, integrating subjective (patient's information) with objective data (biometric information). Our results, although preliminary and if need of further study, appear promising in proposing more effective telemedicine systems. Survey methodology could have an effective role in this growing field of research and applications.
- Published
- 2016
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37. Inhibition of PID1/NYGGF4/PCLI1 gene expression highlights its role in the early events of the cell cycle in NIH3T3 fibroblasts.
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Monteleone F, Vitale M, Caratù G, D'Ambrosio C, Di Giovanni S, Gorrese M, Napolitano F, Romano MF, Del Vecchio L, Succoio M, Scaloni A, and Zambrano N
- Subjects
- Animals, Carrier Proteins biosynthesis, Cell Proliferation genetics, Mice, NIH 3T3 Cells, RNA Interference, RNA, Small Interfering genetics, Structure-Activity Relationship, Carrier Proteins genetics, Carrier Proteins metabolism, Cell Cycle genetics, Down-Regulation genetics, Fibroblasts cytology, Fibroblasts metabolism
- Abstract
The PID1/NYGGF4/PCLI1 gene encodes for a protein with a phosphotyrosine-binding domain, which interacts with the lipoprotein receptor-related protein 1. Previous work by us and others suggested a function of the gene in cell proliferation of NIH3T3 fibroblasts and 3T3-L1 pre-adipocytes. The molecular characterization of PCLI1 protein, ectopically expressed in NIH3T3 fibroblasts, revealed two phosphorylation sites at Ser154 and Ser165. In order to clarify the functions of this gene, we analyzed the effects of its downregulation on cellular proliferation and cell cycle progression in NIH3T3 cell cultures. Downregulation of PID1/NYGGF4/PCLI1 mRNA levels by short hairpin RNAs (shRNAs) elicited decreased proliferation rate in mammalian cell lines; cell cycle analysis of serum-starved, synchronized NIH3T3 fibroblasts showed an increased accumulation of shRNA-interfered cells in the G1 phase. Decreased levels of FOS and MYC mRNAs were accordingly associated with these events. The molecular scenario emerging from our data suggests that PID1/NYGGF4/PCLI1 controls cellular proliferation and cell cycle progression in NIH3T3 cells.
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- 2016
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38. FKBP51 employs both scaffold and isomerase functions to promote NF-κB activation in melanoma.
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Romano S, Xiao Y, Nakaya M, D'Angelillo A, Chang M, Jin J, Hausch F, Masullo M, Feng X, Romano MF, and Sun SC
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- Cell Line, Tumor, Humans, I-kappa B Kinase metabolism, Melanoma enzymology, Protein Interaction Domains and Motifs, TNF Receptor-Associated Factor 2 metabolism, Tacrolimus Binding Proteins chemistry, Melanoma metabolism, NF-kappa B metabolism, Tacrolimus Binding Proteins metabolism
- Abstract
Melanoma is the most aggressive skin cancer; its prognosis, particularly in advanced stages, is disappointing largely due to the resistance to conventional anticancer treatments and high metastatic potential. NF-κB constitutive activation is a major factor for the apoptosis resistance of melanoma. Several studies suggest a role for the immunophilin FKBP51 in NF-κB activation, but the underlying mechanism is still unknown. In the present study, we demonstrate that FKBP51 physically interacts with IKK subunits, and facilitates IKK complex assembly. FKBP51-knockdown inhibits the binding of IKKγ to the IKK catalytic subunits, IKK-α and -β, and attenuates the IKK catalytic activity. Using FK506, an inhibitor of the FKBP51 isomerase activity, we found that the IKK-regulatory role of FKBP51 involves both its scaffold function and its isomerase activity. Moreover, FKBP51 also interacts with TRAF2, an upstream mediator of IKK activation. Interestingly, both FKBP51 TPR and PPIase domains are required for its interaction with TRAF2 and IKKγ, whereas only the TPR domain is involved in interactions with IKKα and β. Collectively, these results suggest that FKBP51 promotes NF-κB activation by serving as an IKK scaffold as well as an isomerase. Our findings have profound implications for designing novel melanoma therapies based on modulation of FKBP51., (© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2015
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39. miR-23a, miR-24 and miR-27a protect differentiating ESCs from BMP4-induced apoptosis.
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Musto A, Navarra A, Vocca A, Gargiulo A, Minopoli G, Romano S, Romano MF, Russo T, and Parisi S
- Subjects
- Animals, Apoptosis drug effects, Apoptosis genetics, Cell Differentiation drug effects, Cell Differentiation genetics, Cell Line, Embryonic Stem Cells drug effects, Mice, MicroRNAs genetics, Bone Morphogenetic Protein 4 pharmacology, Embryonic Stem Cells cytology, Embryonic Stem Cells metabolism, MicroRNAs physiology
- Abstract
Numerous studies have indicated that BMP4 signaling is involved in the regulation of the early steps of development. In mouse embryonic stem cells (ESCs), BMP4 is crucial to sustain pluripotency and blocks differentiation towards neural fate. Here, through a systematic analysis of miRNAs in ESCs, we establish that BMP4 signaling regulates miR-23a, 27a and 24-2, through the recruitment of phospho-Smads at the promoter of the gene encoding this miRNA cluster. Suppression of miR-23a/b, 27a/b and 24 does not affect self-renewal or pluripotency, but induces an evident change of ESC differentiation, with a significant increase of the cells undergoing apoptosis after the transition from ESCs to epiblast stem cells (EpiSCs). BMP4 has been previously reported to cause apoptosis during ESC differentiation. By blocking BMP4 signaling, we completely prevent the apoptosis induced by suppression of the miRs. This suggests that the effects of miR suppression are the result of enhanced BMP4 signaling. This hypothesis is further supported by the observation that Smad5, the transcription factor downstream of the BMP4 receptor, is targeted by the miRNAs of the 23a and 23b clusters. Altogether, our results highlight the existence of a regulatory loop, involving Smad5 and the miR-23a clusters, that modulates the apoptotic response of ESCs to BMP4.
- Published
- 2015
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40. Assessing the carcinogenic potential of low-dose exposures to chemical mixtures in the environment: the challenge ahead.
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Goodson WH 3rd, Lowe L, Carpenter DO, Gilbertson M, Manaf Ali A, Lopez de Cerain Salsamendi A, Lasfar A, Carnero A, Azqueta A, Amedei A, Charles AK, Collins AR, Ward A, Salzberg AC, Colacci A, Olsen AK, Berg A, Barclay BJ, Zhou BP, Blanco-Aparicio C, Baglole CJ, Dong C, Mondello C, Hsu CW, Naus CC, Yedjou C, Curran CS, Laird DW, Koch DC, Carlin DJ, Felsher DW, Roy D, Brown DG, Ratovitski E, Ryan EP, Corsini E, Rojas E, Moon EY, Laconi E, Marongiu F, Al-Mulla F, Chiaradonna F, Darroudi F, Martin FL, Van Schooten FJ, Goldberg GS, Wagemaker G, Nangami GN, Calaf GM, Williams G, Wolf GT, Koppen G, Brunborg G, Lyerly HK, Krishnan H, Ab Hamid H, Yasaei H, Sone H, Kondoh H, Salem HK, Hsu HY, Park HH, Koturbash I, Miousse IR, Scovassi AI, Klaunig JE, Vondráček J, Raju J, Roman J, Wise JP Sr, Whitfield JR, Woodrick J, Christopher JA, Ochieng J, Martinez-Leal JF, Weisz J, Kravchenko J, Sun J, Prudhomme KR, Narayanan KB, Cohen-Solal KA, Moorwood K, Gonzalez L, Soucek L, Jian L, D'Abronzo LS, Lin LT, Li L, Gulliver L, McCawley LJ, Memeo L, Vermeulen L, Leyns L, Zhang L, Valverde M, Khatami M, Romano MF, Chapellier M, Williams MA, Wade M, Manjili MH, Lleonart ME, Xia M, Gonzalez MJ, Karamouzis MV, Kirsch-Volders M, Vaccari M, Kuemmerle NB, Singh N, Cruickshanks N, Kleinstreuer N, van Larebeke N, Ahmed N, Ogunkua O, Krishnakumar PK, Vadgama P, Marignani PA, Ghosh PM, Ostrosky-Wegman P, Thompson PA, Dent P, Heneberg P, Darbre P, Sing Leung P, Nangia-Makker P, Cheng QS, Robey RB, Al-Temaimi R, Roy R, Andrade-Vieira R, Sinha RK, Mehta R, Vento R, Di Fiore R, Ponce-Cusi R, Dornetshuber-Fleiss R, Nahta R, Castellino RC, Palorini R, Abd Hamid R, Langie SA, Eltom SE, Brooks SA, Ryeom S, Wise SS, Bay SN, Harris SA, Papagerakis S, Romano S, Pavanello S, Eriksson S, Forte S, Casey SC, Luanpitpong S, Lee TJ, Otsuki T, Chen T, Massfelder T, Sanderson T, Guarnieri T, Hultman T, Dormoy V, Odero-Marah V, Sabbisetti V, Maguer-Satta V, Rathmell WK, Engström W, Decker WK, Bisson WH, Rojanasakul Y, Luqmani Y, Chen Z, and Hu Z
- Subjects
- Animals, Humans, Carcinogenesis chemically induced, Carcinogens, Environmental adverse effects, Environmental Exposure adverse effects, Hazardous Substances adverse effects, Neoplasms chemically induced, Neoplasms etiology
- Abstract
Lifestyle factors are responsible for a considerable portion of cancer incidence worldwide, but credible estimates from the World Health Organization and the International Agency for Research on Cancer (IARC) suggest that the fraction of cancers attributable to toxic environmental exposures is between 7% and 19%. To explore the hypothesis that low-dose exposures to mixtures of chemicals in the environment may be combining to contribute to environmental carcinogenesis, we reviewed 11 hallmark phenotypes of cancer, multiple priority target sites for disruption in each area and prototypical chemical disruptors for all targets, this included dose-response characterizations, evidence of low-dose effects and cross-hallmark effects for all targets and chemicals. In total, 85 examples of chemicals were reviewed for actions on key pathways/mechanisms related to carcinogenesis. Only 15% (13/85) were found to have evidence of a dose-response threshold, whereas 59% (50/85) exerted low-dose effects. No dose-response information was found for the remaining 26% (22/85). Our analysis suggests that the cumulative effects of individual (non-carcinogenic) chemicals acting on different pathways, and a variety of related systems, organs, tissues and cells could plausibly conspire to produce carcinogenic synergies. Additional basic research on carcinogenesis and research focused on low-dose effects of chemical mixtures needs to be rigorously pursued before the merits of this hypothesis can be further advanced. However, the structure of the World Health Organization International Programme on Chemical Safety 'Mode of Action' framework should be revisited as it has inherent weaknesses that are not fully aligned with our current understanding of cancer biology., (© The Author 2015. Published by Oxford University Press.)
- Published
- 2015
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41. Disruptive environmental chemicals and cellular mechanisms that confer resistance to cell death.
- Author
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Narayanan KB, Ali M, Barclay BJ, Cheng QS, D'Abronzo L, Dornetshuber-Fleiss R, Ghosh PM, Gonzalez Guzman MJ, Lee TJ, Leung PS, Li L, Luanpitpong S, Ratovitski E, Rojanasakul Y, Romano MF, Romano S, Sinha RK, Yedjou C, Al-Mulla F, Al-Temaimi R, Amedei A, Brown DG, Ryan EP, Colacci A, Hamid RA, Mondello C, Raju J, Salem HK, Woodrick J, Scovassi AI, Singh N, Vaccari M, Roy R, Forte S, Memeo L, Kim SY, Bisson WH, Lowe L, and Park HH
- Subjects
- Animals, Homeostasis drug effects, Humans, Carcinogenesis chemically induced, Carcinogens, Environmental adverse effects, Cell Death drug effects, Environmental Exposure adverse effects, Hazardous Substances adverse effects, Neoplasms chemically induced, Neoplasms etiology
- Abstract
Cell death is a process of dying within biological cells that are ceasing to function. This process is essential in regulating organism development, tissue homeostasis, and to eliminate cells in the body that are irreparably damaged. In general, dysfunction in normal cellular death is tightly linked to cancer progression. Specifically, the up-regulation of pro-survival factors, including oncogenic factors and antiapoptotic signaling pathways, and the down-regulation of pro-apoptotic factors, including tumor suppressive factors, confers resistance to cell death in tumor cells, which supports the emergence of a fully immortalized cellular phenotype. This review considers the potential relevance of ubiquitous environmental chemical exposures that have been shown to disrupt key pathways and mechanisms associated with this sort of dysfunction. Specifically, bisphenol A, chlorothalonil, dibutyl phthalate, dichlorvos, lindane, linuron, methoxychlor and oxyfluorfen are discussed as prototypical chemical disruptors; as their effects relate to resistance to cell death, as constituents within environmental mixtures and as potential contributors to environmental carcinogenesis., (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2015
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42. FKBP51 increases the tumour-promoter potential of TGF-beta.
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Romano S, D'Angelillo A, D'Arrigo P, Staibano S, Greco A, Brunetti A, Scalvenzi M, Bisogni R, Scala I, and Romano MF
- Abstract
Background: FKBP51 (FKBP5 Official Symbol) is a large molecular weight component of the family of FK506 binding proteins (FKBP). In recent years, research studies from our laboratory highlighted functions for FKBP51 in the control of apoptosis and melanoma progression. FKBP51 expression correlated with the invasiveness and aggressiveness of melanoma. Since a role for TGF-β in the enhanced tumorigenic potential of melanoma cells is widely described, we hypothesized a cooperative effect between FKBP51 and TGF-β in melanoma progression., Methods: SAN and A375 melanoma cell lines were utilized for this study. Balb/c IL2γ NOD SCID served to assess the ability to colonize organs and metastasize of different cell lines, which was evaluated by in vivo imaging. Realtime PCR and western blot served for measurement of mRNA and protein expression, respectively., Results: By comparing the metastatic potential of two melanoma cell lines, namely A375 and SAN, we confirmed that an increased capability to colonize murine organs was associated with increased levels of FKBP51. A375 melanoma cell line expressed FKBP51 mRNA levels 30-fold higher in comparison to the SAN mRNA level and appeared more aggressive than SAN melanoma cell line in an experimental metastasis model. In addition, A375 expressed, more abundantly than SAN, the TGF-β and the pro angiogenic TGF-β receptor type III (TβRIII) factors. FKBP51 silencing produced a reduction of TGF-β and TβRIII gene expression in A375 cell line, in accordance with previous studies. We found that the inducing effect of TGF-β on Sparc and Vimentin expression was impaired in condition of FKBP51 depletion, suggesting that FKBP51 is an important cofactor in the TGF-β signal. Such a hypothesis was supported by co immunoprecipitation assays, showing that FKBP51 interacted with either Smad2,3 and p300. In normal melanocytes, FKBP51 potentiated the effect of TGF-β on N-cadherin expression and conferred a mesenchymal-like morphology to such round-shaped cells., Conclusions: Overall, our findings show that FKBP51 enhances some pro oncogenic functions of TGF-β, suggesting that FKBP51-overexpression may help melanoma to take advantage of the tumor promoting activities of the cytokine.
- Published
- 2014
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43. Translational control in the stress adaptive response of cancer cells: a novel role for the heat shock protein TRAP1.
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Matassa DS, Amoroso MR, Agliarulo I, Maddalena F, Sisinni L, Paladino S, Romano S, Romano MF, Sagar V, Loreni F, Landriscina M, and Esposito F
- Subjects
- Colorectal Neoplasms genetics, Down-Regulation, Endoplasmic Reticulum Chaperone BiP, Eukaryotic Initiation Factor-2 metabolism, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, HCT116 Cells, Humans, Neoplasms metabolism, Neoplasms pathology, Protein Binding, Proteolysis, Ribosomes metabolism, Signal Transduction, Ubiquitination, Colorectal Neoplasms metabolism, Colorectal Neoplasms pathology, Heat-Shock Proteins metabolism, Protein Biosynthesis, Stress, Physiological, TNF Receptor-Associated Factor 1 metabolism
- Abstract
TNF receptor-associated protein 1 (TRAP1), the main mitochondrial member of the heat shock protein (HSP) 90 family, is induced in most tumor types and is involved in the regulation of proteostasis in the mitochondria of tumor cells through the control of folding and stability of selective proteins, such as Cyclophilin D and Sorcin. Notably, we have recently demonstrated that TRAP1 also interacts with the regulatory protein particle TBP7 in the endoplasmic reticulum (ER), where it is involved in a further extra-mitochondrial quality control of nuclear-encoded mitochondrial proteins through the regulation of their ubiquitination/degradation. Here we show that TRAP1 is involved in the translational control of cancer cells through an attenuation of global protein synthesis, as evidenced by an inverse correlation between TRAP1 expression and ubiquitination/degradation of nascent stress-protective client proteins. This study demonstrates for the first time that TRAP1 is associated with ribosomes and with several translation factors in colon carcinoma cells and, remarkably, is found co-upregulated with some components of the translational apparatus (eIF4A, eIF4E, eEF1A and eEF1G) in human colorectal cancers, with potential new opportunities for therapeutic intervention in humans. Moreover, TRAP1 regulates the rate of protein synthesis through the eIF2α pathway either under basal conditions or under stress, favoring the activation of GCN2 and PERK kinases, with consequent phosphorylation of eIF2α and attenuation of cap-dependent translation. This enhances the synthesis of selective stress-responsive proteins, such as the transcription factor ATF4 and its downstream effectors BiP/Grp78, and the cystine antiporter system xCT, thereby providing protection against ER stress, oxidative damage and nutrient deprivation. Accordingly, TRAP1 silencing sensitizes cells to apoptosis induced by novel antitumoral drugs that inhibit cap-dependent translation, such as ribavirin or 4EGI-1, and reduces the ability of cells to migrate through the pores of transwell filters. These new findings target the TRAP1 network in the development of novel anti-cancer strategies.
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- 2013
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44. Somatic overgrowth predisposes to seizures in autism spectrum disorders.
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Valvo G, Baldini S, Brachini F, Apicella F, Cosenza A, Ferrari AR, Guerrini R, Muratori F, Romano MF, Santorelli FM, Tancredi R, and Sicca F
- Subjects
- Adolescent, Behavior, Child, Child, Preschool, Cognition, Family, Female, Humans, Language Development, Male, Young Adult, Body Size, Child Development Disorders, Pervasive complications, Disease Susceptibility complications, Seizures complications
- Abstract
Background: Comorbidity of Autism Spectrum Disorders with seizures or abnormal EEG (Autism-Epilepsy Phenotype) suggests shared pathomechanisms, and might be a starting point to identify distinct populations within the clinical complexity of the autistic spectrum. In this study, we tried to assess whether distinct subgroups, having distinctive clinical hallmarks, emerge from this comorbid condition., Methods: Two-hundred and six individuals with idiopathic Autism Spectrum Disorders were subgrouped into three experimental classes depending on the presence of seizures and EEG abnormalities. Neurobehavioral, electroclinical and auxological parameters were investigated to identify differences among groups and features which increase the risk of seizures. Our statistical analyses used ANOVA, post-hoc multiple comparisons, and the Chi-squared test to analyze continuous and categorical variables. A correspondence analysis was also used to decompose significant Chi-squared and reduce variables dimensions., Results: The high percentage of children with seizures (28.2% of our whole cohort) and EEG abnormalities (64.1%) confirmed that the prevalence of epilepsy in Autism Spectrum Disorders exceeds that of the general population. Seizures were associated with severe intellectual disability, and not with autism severity. Interestingly, tall stature (without macrocephaly) was significantly associated with EEG abnormalities or later onset seizures. However, isolated macrocephaly was equally distributed among groups or associated with early onset seizures when accompanied by tall stature., Conclusions: Tall stature seems to be a phenotypic "biomarker" of susceptibility to EEG abnormalities or late epilepsy in Autism Spectrum Disorders and, when concurring with macrocephaly, predisposes to early onset seizures. Growth pattern might act as an endophenotypic marker in Autism-Epilepsy comorbidity, delineating distinct pathophysiological subtypes and addressing personalized diagnostic work-up and therapeutic approaches.
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- 2013
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45. FK506 binding protein 51 positively regulates melanoma stemness and metastatic potential.
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Romano S, Staibano S, Greco A, Brunetti A, Nappo G, Ilardi G, Martinelli R, Sorrentino A, Di Pace A, Mascolo M, Bisogni R, Scalvenzi M, Alfano B, and Romano MF
- Subjects
- Animals, Biomarkers metabolism, Cell Line, Tumor, Cell Movement, Epithelial-Mesenchymal Transition genetics, Humans, Intermediate Filament Proteins genetics, Intermediate Filament Proteins metabolism, Melanoma genetics, Melanoma metabolism, Mice, Neoplasm Invasiveness, Neoplasms, Experimental, Neoplastic Stem Cells pathology, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, Nestin, RNA, Small Interfering genetics, Signal Transduction, Skin Neoplasms genetics, Skin Neoplasms metabolism, Tacrolimus Binding Proteins antagonists & inhibitors, Tacrolimus Binding Proteins metabolism, Gene Expression Regulation, Neoplastic, Melanoma pathology, Neoplastic Stem Cells metabolism, Skin Neoplasms pathology, Tacrolimus Binding Proteins genetics
- Abstract
Melanoma is the most aggressive skin cancer; there is no cure in advanced stages. Identifying molecular participants in melanoma progression may provide useful diagnostic and therapeutic tools. FK506 binding protein 51 (FKBP51), an immunophilin with a relevant role in developmental stages, is highly expressed in melanoma and correlates with aggressiveness and therapy resistance. We hypothesized a role for FKBP51 in melanoma invasive behaviour. FKBP51 promoted activation of epithelial-to-mesenchymal transition (EMT) genes and improved melanoma cell migration and invasion. In addition, FKBP51 induced some melanoma stem cell (MCSC) genes. Purified MCSCs expressed high EMT genes levels, suggesting that genetic programs of EMT and MCSCs overlap. Immunohistochemistry of samples from patients showed intense FKBP51 nuclear signal and cytoplasmic positivity for the stem cell marker nestin in extravasating melanoma cells and metastatic brains. In addition, FKBP51 targeting by small interfering RNA (siRNA) prevented the massive metastatic substitution of liver and lung in a mouse model of experimental metastasis. The present study provides evidence that the genetic programs of cancer stemness and invasiveness overlap in melanoma, and that FKBP51 plays a pivotal role in sustaining such a program.
- Published
- 2013
- Full Text
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46. Synergy between direct coronary stenting technique and use of the novel thin strut cobalt chromium Skylor™ stent: the MACE in follow up patients treated with Skylor stent [MILES Study].
- Author
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Giordano A, Polimeno M, Corcione N, Fattore L, Di Lorenzo L, Biondi-Zoccai G, Ferraro P, and Romano MF
- Subjects
- Aged, Chromium Alloys therapeutic use, Female, Humans, Kaplan-Meier Estimate, Male, Middle Aged, Percutaneous Coronary Intervention methods, Prosthesis Design, Retrospective Studies, Stents statistics & numerical data, Treatment Outcome, Coronary Restenosis etiology, Myocardial Infarction etiology, Percutaneous Coronary Intervention instrumentation, Stents adverse effects
- Abstract
Background: Despite significant improvements in stent platform, currently available bare-metal stents (BMS) are still associated with restenosis. Thin-strut design cobalt-chromium alloys hold the promise of improving results of BMS, especially when implanted with direct technique. We performed an observational study to appraise outcomes of the novel Skylor™ stent, stratifying outcomes according to stenting technique., Methods and Results: We included all consecutive patients undergoing coronary stenting with Skylor™ at 2 centers between 2006 and 2009. The primary end-point was the long-term rate of major adverse cardiac events (MACE, i.e. death, myocardial infarction (MI), coronary artery bypass grafting (CABG) or target vessel revascularization (TVR)). As pre-specified analysis, we compared patients undergoing direct stenting versus those stent implantation following predilation. A total of 1020 patients were included (1292 Skylor™ stents), with procedural success obtained in 99%. Comparing patients undergoing direct stenting (66%) versus pre-dilation (34%) at 16±7 months of follow-up, MACE had occurred in, respectively, 8% versus 14% (p=0.001), with death in 1% versus 2= (p=0.380), MI in 1% versus 2% (p=0.032), CABG in 0.2% versus 2% (p=0.012), and TVR in 6% versus 9% [p=0.071]. Even at multivariable analysis with propensity adjustment, direct stenting was associated with significantly fewer MACE [hazard ratio 0.60 [0.38-0.93], p=0.024]., Conclusions: This observational study suggests the presence of a beneficial synergy between direct coronary stenting technique and use of the novel thin-strut cobalt-chromium Skylor™ stent in real-world patients undergoing PCI.
- Published
- 2012
- Full Text
- View/download PDF
47. Differential effect of atorvastatin and tacrolimus on proliferation of vascular smooth muscle and endothelial cells.
- Author
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Giordano A, Romano S, Monaco M, Sorrentino A, Corcione N, Di Pace AL, Ferraro P, Nappo G, Polimeno M, and Romano MF
- Subjects
- Aged, Aged, 80 and over, Antigens, CD metabolism, Atorvastatin, Cardiovascular Agents adverse effects, Cells, Cultured, Cyclin B metabolism, DNA Replication drug effects, Dose-Response Relationship, Drug, Drug-Eluting Stents, Endoglin, Human Umbilical Vein Endothelial Cells metabolism, Humans, Male, Mitogen-Activated Protein Kinase 1 metabolism, Mitogen-Activated Protein Kinase 3 metabolism, Muscle, Smooth, Vascular metabolism, Myocytes, Smooth Muscle metabolism, Phosphorylation, Receptors, Cell Surface metabolism, Tacrolimus adverse effects, Time Factors, beta Catenin metabolism, Cardiovascular Agents pharmacology, Cell Proliferation drug effects, Heptanoic Acids pharmacology, Human Umbilical Vein Endothelial Cells drug effects, Hydroxymethylglutaryl-CoA Reductase Inhibitors pharmacology, Muscle, Smooth, Vascular drug effects, Myocytes, Smooth Muscle drug effects, Pyrroles pharmacology, Tacrolimus pharmacology
- Abstract
Although considered promising for use in drug-eluting stents (DES), tacrolimus failed clinically. Tacrolimus inhibits growth factor production but can also act as a growth factor on vascular smooth muscle cells (VSMC). This unexpected proliferative stimulus could reverse the beneficial effects of the drug on restenosis. We hypothesized that tacrolimus' association with statins, which lower cholesterol and impair cell proliferation, could restore tacrolimus' beneficial effect by abrogating the aberrant proliferative stimulus. Additionally, since maintenance of endothelial function represents a challenge for new-generation DES, we investigated the combined effect of tacrolimus and atorvastatin on endothelial cells. Human VSMC and umbilical vein endothelial cells (HUVEC) were incubated with 100 nM tacrolimus and increasing doses of atorvastatin (0-3.0 μM). Atorvastatin plus tacrolimus dose-dependently inhibited VSMC proliferation. The percentage of cells incorporating 5-bromo-2'-deoxyuridine (BrdU) in their DNA was 49 ± 14% under basal conditions, 62 ± 15% (P = 0.01) with tacrolimus, 40 ± 22% with 3 μM atorvastatin, and 30 ± 7% (P < 0.05) with 3 μM atorvastatin plus tacrolimus. Atorvastatin downregulated β-catenin, Erk1 and Erk2, and cyclin B in tacrolimus-stimulated VSMC. In contrast, atorvastatin plus tacrolimus did not affect proliferation of endothelial cells. The percentage of HUVEC incorporating BrdU in their DNA was 47 ± 8% under basal conditions, 58 ± 6% (P = 0.01) with tacrolimus, 45 ± 4% with 3 μM atorvastatin, and 49 ± 1% with 3 μM atorvastatin plus tacrolimus. Both agents stimulated endoglin production by HUVEC. Taken together, these results suggest that, when combined with tacrolimus, atorvastatin exerts a dose-dependent antiproliferative effect on VSMC. In contrast, atorvastatin acts in concert with tacrolimus in HUVEC to stimulate production of endoglin, a factor that has an important role in endothelial repair. Our study supports the conclusion that prevention of postcoronary in-stent restenosis and late thrombosis may benefit of concomitant association of tacrolimus and high doses of atorvastatin.
- Published
- 2012
- Full Text
- View/download PDF
48. In vivo and in vitro assessment of pathways involved in contrast media-induced renal cells apoptosis.
- Author
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Quintavalle C, Brenca M, De Micco F, Fiore D, Romano S, Romano MF, Apone F, Bianco A, Zabatta MA, Troncone G, Briguori C, and Condorelli G
- Subjects
- Acute Kidney Injury chemically induced, Adult, Aged, Aged, 80 and over, Caspase 3 metabolism, Cells, Cultured, Enzyme Assays, Female, Humans, JNK Mitogen-Activated Protein Kinases metabolism, Kidney Tubules drug effects, Male, Middle Aged, Reactive Oxygen Species metabolism, bcl-2 Homologous Antagonist-Killer Protein metabolism, Acute Kidney Injury pathology, Apoptosis drug effects, Contrast Media adverse effects, Kidney Tubules pathology, Signal Transduction drug effects
- Abstract
Contrast-induced nephropathy accounts for >10% of all causes of hospital-acquired renal failure, causes a prolonged in-hospital stay and represents a powerful predictor of poor early and late outcome. Mechanisms of contrast-induced nephropathy are not completely understood. In vitro data suggests that contrast media (CM) induces a direct toxic effect on renal tubular cells through the activation of the intrinsic apoptotic pathway. It is unclear whether this effect has a role in the clinical setting. In this work, we evaluated the effects of CM both in vivo and in vitro. By analyzing urine samples obtained from patients who experienced contrast-induced acute kidney injury (CI-AKI), we verified, by western blot and immunohistochemistry, that CM induces tubular renal cells apoptosis. Furthermore, in cultured cells, CM caused a dose-response increase in reactive oxygen species (ROS) production, which triggered Jun N-terminal kinases (JNK1/2) and p38 stress kinases marked activation and thus apoptosis. Inhibition of JNK1/2 and p38 by different approaches (i.e. pharmacological antagonists and transfection of kinase-death mutants of the upstream p38 and JNK kinases) prevented CM-induced apoptosis. Interestingly, N-acetylcysteine inhibited ROS production, and thus stress kinases and apoptosis activation. Therefore, we conclude that CM-induced tubular renal cells apoptosis represents a key mechanism of CI-AKI.
- Published
- 2011
- Full Text
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49. FK506-binding protein 51 is a possible novel tumoral marker.
- Author
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Romano S, D'Angelillo A, Staibano S, Ilardi G, and Romano MF
- Subjects
- Humans, Neoplasms metabolism, Proto-Oncogene Proteins c-akt metabolism, RNA, Messenger metabolism, Tacrolimus Binding Proteins genetics, Biomarkers, Tumor metabolism, Tacrolimus Binding Proteins metabolism
- Published
- 2010
- Full Text
- View/download PDF
50. Role of FK506-binding protein 51 in the control of apoptosis of irradiated melanoma cells.
- Author
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Romano S, D'Angelillo A, Pacelli R, Staibano S, De Luna E, Bisogni R, Eskelinen EL, Mascolo M, Calì G, Arra C, and Romano MF
- Subjects
- Animals, Apoptosis Regulatory Proteins metabolism, Beclin-1, Cell Line, Tumor, Humans, Melanoma metabolism, Melanoma pathology, Mice, Mice, Nude, NF-kappa B metabolism, RNA, Small Interfering metabolism, Tacrolimus Binding Proteins genetics, Tacrolimus Binding Proteins metabolism, Transplantation, Heterologous, X-Linked Inhibitor of Apoptosis Protein metabolism, bcl-2-Associated X Protein metabolism, Apoptosis, Melanoma radiotherapy, Radiation, Ionizing, Tacrolimus Binding Proteins physiology
- Abstract
FK506-binding protein 51 (FKBP51) is an immunophilin with isomerase activity, which performs important biological functions in the cell. It has recently been involved in the apoptosis resistance of malignant melanoma. The aim of this study was to investigate the possible role of FKBP51 in the control of response to ionizing radiation (Rx) in malignant melanoma. FKBP51-silenced cells showed reduced clonogenic potential after irradiation compared with non-silenced cells. After Rx, we observed apoptosis in FKBP51-silenced cells and autophagy in non-silenced cells. The FKBP51-controlled radioresistance mechanism involves NF-kappaB. FKBP51 was required for the activation of Rx-induced NF-kappaB, which in turn inhibited apoptosis by stimulating X-linked inhibitor of apoptosis protein and promoting authophagy-mediated Bax degradation. Using a tumor-xenograft mouse model, the in vivo pretreatment of tumors with FKBP51-siRNA provoked massive apoptosis after irradiation. Immunohistochemical analysis of 10 normal skin samples and 80 malignant cutaneous melanomas showed that FKBP51 is a marker of melanocyte malignancy, correlating with vertical growth phase and lesion thickness. Finally, we provide evidence that FKBP51 targeting radiosensitizes cancer stem/initiating cells. In conclusion, our study identifies a possible molecular target for radiosensitizing therapeutic strategies against malignant melanoma.
- Published
- 2010
- Full Text
- View/download PDF
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