76 results on '"Cappabianca L"'
Search Results
2. TRKAIII COMMUNICATES ER-STRESS TO THE MITO-CHONDRIA IN NEUROBLASTOMA CELLS, RESULTING INGLYCOLYTIC METABOLIC ADAPTATION
- Author
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Farina, A. R., Cappabianca, L., Ruggeri, P., Gneo, L., and Mackay, A. R.
- Subjects
TRKA III - Published
- 2018
3. Advanced Merkel cell carcinoma of the lower extremity treated with surgery and isolated pelvic and limb perfusion using Melphalan: A case of unexpected long-term survival
- Author
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Guadagni, S., primary, Chiominto, A., additional, Mackay, A.R., additional, Farina, A.R., additional, Cappabianca, L., additional, Puccica, I., additional, Valiyeva, S., additional, and Clementi, M., additional
- Published
- 2019
- Full Text
- View/download PDF
4. Stigma di genere e prevenzione del bullismo omofobico
- Author
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AMODEO, ANNA LISA, VALERIO, PAOLO, Cascone M., Garzillo F., Cappabianca L., Di Lello E., Amodeo, ANNA LISA, Cascone, M., Garzillo, F., Cappabianca, L., Di Lello, E., and Valerio, Paolo
- Subjects
prevenzione ,insegnanti ,stigma di genere - Abstract
Il bullismo omofobico costituisce una forma di discriminazione che utilizza un gergo connotato sessualmente, rivolto non soltanto a giovani LGBT ma anche a tutte quelle persone che vengono percepite come non aderenti agli stereotipi sessuali normativi. Le forme con cui può essere messo in atto sono molteplici e vanno dall’esclusione, all’isolamento, alla minaccia, fino ad arrivare a vere e proprie forme di violenze ed aggressioni, agite in nome dell’omofobia, del sessismo e dei valori associati all’eterosessismo. Il rischio di sviluppare nel medio e lungo termine disturbi di tipo internalizing in chi ripetutamente subisce questo tipo di prevaricazione è elevato. Risulta, pertanto, necessario attuare interventi di prevenzione, sensibilizzazione e formazione ai temi del rispetto delle differenze di genere, soprattutto in quei contesti che hanno come obiettivo l’educazione e lo sviluppo psicosociale della persona. La scuola innanzitutto, che può costituire una culla di sviluppo dell’omofobia e delle prevaricazioni legate allo stigma di genere. Scopo del presente contributo è quello di descrivere l’intervento svolto con un gruppo di docenti, all’interno di un più ampio progetto di prevenzione del bullismo omofobico. L’intervento, consistito in un ciclo di 6 incontri di discussione di gruppo, ha avuto come risultato quello di rendere i docenti maggiormente capaci di supportare i propri allievi nel processo di autodeterminazione del proprio orientamento sessuale e, pertanto, di tentare di ridurre il rischio di subire prevaricazioni omofobiche.
- Published
- 2010
5. NGF FLIPs TrkA onto the death TRAIL in neuroblastoma cells
- Author
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Ruggeri, P, primary, Cappabianca, L, additional, Farina, A R, additional, Gneo, L, additional, and Mackay, A R, additional
- Published
- 2016
- Full Text
- View/download PDF
6. NGF sensitizes TrkA SH-SY5Y neuroblastoma cells to TRAIL-induced apoptosis
- Author
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Ruggeri, P, primary, Cappabianca, L, additional, Farina, A R, additional, Gneo, L, additional, and Mackay, A R, additional
- Published
- 2016
- Full Text
- View/download PDF
7. The alternative TrkAIII splice variant expressed by human neuroblastomas and glioblastomas targets the centrosome and promotes genetic instability
- Author
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Farina, Ar, Tacconelli, A, Cappabianca, L, and Mackay, ANDREW REAY
- Published
- 2009
8. A NOVEL HYPOXIA-REGULATED ALTERNATIVE TRKA SPLICE VARIANT AF POTENTIAL PHYSIOLOGICAL AND PATHOLOGICAL IMPORTANCE
- Author
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Tacconelli, A., Farina, Antonietta, Cappabianca, L., Gulino, A., and Mackay, A. R.
- Published
- 2005
9. RECONSTITUTION OF TIMP-2 EXPRESSION IN SH-SY5Y NEUROBLASTOMA CELL BY 5-AZACYTIDINA IS MEDIATED TRANSCRIPTIONALLY BY NF-Y THROUGH AN INVERTED CAAT SITE
- Author
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Cappabianca, L, Farina, Antonietta, Tacconelli, A, Mantovani, R, Gulino, A, and Mackay, A. R.
- Published
- 2003
10. TIOREDOXIN INHIBITS MICROVASCULAR ENDHOTHELIAL CAPILLARY TUBULE FORMATION
- Author
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Farina, Antonietta, Tacconelli, A, Cappabianca, L, DE SANTIS, G, Gulino, A, and Mackay, A.
- Published
- 2003
11. Al trans Retinoic Acid Induces NF-kB activation, Matrix Metalloproteinase-9 expression and Enhances Basement Membrane Invasivity of Differentiation Resistant Human SK-N-BE 9N Neuroblastoma Cells
- Author
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Farina, A. R., Masciulli, Mp, Tacconelli, A, Cappabianca, L, DE SANCTIS, G, Gulino, Alberto, and Mackay, A. R.
- Published
- 2002
12. ALL-TRANS-RETINOIC INDUCES NUCLEAR FACTOR-B ACTIVATION AND MATRIX METALLOPROTEINASE-9 EXPRESSION AND ENHANCES BASEMENT MEMBRANE INVASIVITY OF DIFFERENTIATION-RESISTANT HUMAN SK-N-BE 9N NEUROBLASTOMA CELL
- Author
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Farina, Antonietta, Masciulli, M. P., Tacconelli, A., Cappabianca, L., DE SANTIS, G., Gulino, A., and Mackay, A. R.
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- 2002
13. All-trans-retinoic acid induces nuclear factor kappa B activation and matrix metalloproteinase-9 expression and enhances basement membrane invasivity of differentiation-resistant human SK-N-BE 9N neuroblastoma cells
- Author
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Ar, Farina, Mp, Masciulli, Tacconelli A, Cappabianca L, De Santis G, Gulino A, and Andrew Reay Mackay
- Published
- 2002
14. Thioredoxin alters matrix metalloproteinase/tissue inhibitors of metalloproteinase balance and stimulates human SK-N-SH neuroblastoma cell invasion
- Author
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Farina, Ar, Tacconelli, A., Cappabianca, L., Masciulli, Mp, Holmgren, A., Beckett, Gj, Gulino, Alberto, and Mackay, A. R.
- Published
- 2000
15. RETINOIC ACID ENHANCES INVASION THROUGH RECONSTITUTED BASEMENT MEMBRANE BY HUMAN SK-N-SH NEUROBLASTOMA CELLS INVOLVES MEMBRANE-ASSOCIATED TISSUE TYPE PLASMINOGEN ACTIVATOR
- Author
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Tiberio, A, Farina, Ar, Tacconelli, A, Cappabianca, L, Gulino, A, and Mackay, ANDREW REAY
- Published
- 1997
16. Identification of plasminogen in Matrigel and its activation by plasminogen activator endogenous to this tumor basement membrane extract
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Farina, A. R., Tiberio, A., Tacconelli, A., Cappabianca, L., Gulino, Alberto, and Mackay, A. R.
- Published
- 1996
17. Follicle stimulating hormone increases the expression of tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2 and induces TIMP-1 AP-1 site binding complexes in prepubertal rat Sertoli cells
- Author
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Ulisse, Salvatore, Farina, Ar, Piersanti, D, Tiberio, A, Cappabianca, L, D’Orazi, G, Jannini, Ea, Maljk, O, Stetler Stevenson WG, D'Armiento, Massimino, Gulino, Alberto, and Mackay, Ar
- Subjects
TIMP-2 ,TIMP-1 ,Sertoli cells ,FSH ,AP-1 binding site - Published
- 1995
18. Identification of Plasminogen in MatrigelTM and Its Activation by Reconstitution of This Basement Membrane Extract
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Farina, A.R., primary, Tiberio, A., additional, Tacconelli, A., additional, Cappabianca, L., additional, Gulino, A., additional, and Mackay, A.R., additional
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- 1996
- Full Text
- View/download PDF
19. Follicle-stimulating hormone increases the expression of tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2 and induces TIMP-1 AP-1 site binding complex(es) in prepubertal rat Sertoli cells.
- Author
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Ulisse, S, primary, Farina, A R, additional, Piersanti, D, additional, Tiberio, A, additional, Cappabianca, L, additional, D'Orazi, G, additional, Jannini, E A, additional, Malykh, O, additional, Stetler-Stevenson, W G, additional, and D'Armiento, M, additional
- Published
- 1994
- Full Text
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20. Positive and negative regulation of the composite octamer motif of the interleukin 2 enhancer by AP-1, Oct-2, and retinoic acid receptor.
- Author
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de Grazia, U, primary, Felli, M P, additional, Vacca, A, additional, Farina, A R, additional, Maroder, M, additional, Cappabianca, L, additional, Meco, D, additional, Farina, M, additional, Screpanti, I, additional, Frati, L, additional, and Gulino, A, additional
- Published
- 1994
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21. Frequency response of thin film heads with longitudinal and transverse anisotropy
- Author
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Mallary, M., primary, Torabi, A., additional, Batra, S., additional, Smith, A.B., additional, Cappabianca, L., additional, and Goller, W.W., additional
- Published
- 1990
- Full Text
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22. Identification of Plasminogen in MatrigelTMand Its Activation by Reconstitution of This Basement Membrane Extract
- Author
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Farina, A.R., Tiberio, A., Tacconelli, A., Cappabianca, L., Gulino, A., Gulino, A., and Mackay, A.R.
- Abstract
MatrigelTM, a basement membrane (BM) extract of the Engelbreth-Holm-Swarm (EHS) sarcoma, used in tumor invasion assays, was found to contain plasminogen. Plasminogen was identified, using Western blot analysis and casein zymograms, by comparison with human plasminogen. Matrigel contained approximately 20–100 ng of plasminogen per 100 μg of protein as determined by these assays. Matrigel reconstitution and incubation at 37°C caused activation of plasminogen, which was serine protease dependent and involved tissue plasminogen activator (tPA) as an anti-tPA antibody which inhibited activation. This reconstitution and incubation also caused leupeptin-inhibitable degradation of Matrigel components as assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Degradation of the BM extract copolymerized in zymograms was caused by human plasminogen and plasminogen in the Matrigel. Maximal plasmin activity, following incubation of Matrigel at 37°C for 16 h, was equivalent to approximately 10 ng of purified plasmin using the plasmin substrate d-Val-Leu-Lys p-nitroanilide. Matrigel, therefore, contained all the components of the plasmin-generating system, including plasminogen. The plasmin generated degraded Matrigel components and exogenous substrates. Our data suggest that, since this tumor BM acts as a reservoir for enzymes of the plasmin-generating system, caution should be taken by investigators interpreting data concerning the effects of Matrigel on cell behavior and, in particular, cellular invasion.
- Published
- 1996
- Full Text
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23. Transcriptional regulation of intercellular adhesion molecule 1 by phorbol ester in human neuroblastoma cell line SK-N-SH involves jun- and fos-containing activator protein 1 site binding complex(es)
- Author
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Antonietta Rosella FARINA, Cappabianca, L., Mackay, A. R., Tiberio, A., Tacconelli, A., Tessitore, A., Frati, L., Martinotti, S., and Gulino, A.
- Subjects
TUMOR-NECROSIS-FACTOR ,HUMAN-ENDOTHELIAL-CELLS ,RETINOIC ACID ,ICAM-1 EXPRESSION ,DNA-BINDING ,INTERFERON-GAMMA ,FACTOR-ALPHA ,C-FOS ,INFLAMMATORY CYTOKINES ,GENE-TRANSCRIPTION ,Transcriptional Activation ,Transcription, Genetic ,Proto-Oncogene Proteins c-jun ,Regulatory Sequences, Nucleic Acid ,Neuroblastoma ,Humans ,RNA, Messenger ,Cyclic AMP Response Element-Binding Protein ,NF-kappa B ,DNA, Neoplasm ,Intercellular Adhesion Molecule-1 ,TATA Box ,Gene Expression Regulation, Neoplastic ,Transcription Factor AP-1 ,Tetradecanoylphorbol Acetate ,Proto-Oncogene Proteins c-fos ,Protein Binding - Abstract
In this study, the regulatory elements involved in ICAM-1 transcriptional response to phorbol ester (12-0-tetradecanoylphorbol-13-acetate; TPA) have been investigated in the human neuroblastoma cell line, SK-N-SH. TPA induced intercellular adhesion molecule 1 (ICAM-1) protein expression in SK-N-SH cells within 24 h of treatment as judged by indirect immunofluorescence. Basal ICAM-1 mRNA levels were barely detectable in untreated SK-N-SH cells but were induced by TPA to a maximal level with 4 h and were reduced thereafter. Analysis of the 5' promoter sequence of ICAM-1 revealed two regions that functioned equally in the TPA induction of ICAM-1 transcription. The first region (-145 to -227) contained a nuclear factor-kappa B (NF kappa B) element. The second region (-316 to -390) contained a putative TPA-responsive element (TRE; TGATTCA) and a TATA box. Deletion and point mutation of the latter region indicated that the TRE was indeed the functional element within this region and acted fully and independently of all other elements including the TATA box at position -352. This TRE bound TPA induced specific nuclear complexes in vitro containing junD, c-jun, c-fos, and fra2 but not cAMP-responsive element binding/activating transcription factor family proteins. ICAM-TRE binding activity was induced within 30 min following TPA treatment. This preceded the appearance of ICAM-NF kappa B site binding activity. Cotransfection of c-jun and c-fos expression vectors into SK-N-SH cells induced transactivation from ICAM-1 promoter constructs containing the intact but not mutated TRE site. Primer extension analyses revealed that TPA had induced transcription exclusively at two sites -40 and -41 bp upstream of the translation start site. These data show that the ICAM-TRE and its cognate jun- and fos-containing transcription factors play a predominant role in the transcriptional response of ICAM-1 to the protein kinase C activator TPA in SK-N-SH cells.
24. Identification of plasminogen in Matrigel(TM) and its activation by reconstitution of this basement membrane extract
- Author
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Antonietta Rosella FARINA, Tiberio, A., Tacconelli, A., Cappabianca, L., Gulino, A., and Mackay, A. R.
25. Follicle-Stimulating Hormone Increases the Expression of Tissue Inhibitors of Metalloproteinases TIMP-1 and TIMP-2 and Induces TIMP-1 AP-1 Site Binding Complex(es) in Prepubertal Rat Sertoli Cells
- Author
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Ulisse, S., Farina, A. R., Piersanti, D., Tiberio, A., Cappabianca, L., D Orazi, G., Jannini, E. A., Malykh, O., William Stetler-Stevenson, D Armiento, M., Gulino, A., and Mackay, A. R.
- Subjects
Male ,Proto-Oncogene Proteins c-jun ,FOS TRANSCRIPTION ,Matrix metalloproteinase ,chemistry.chemical_compound ,Follicle-stimulating hormone ,Endocrinology ,Sexual Maturation ,Cyclic AMP Response Element-Binding Protein ,Cells, Cultured ,APE LEUKEMIA-VIRUS ,Metalloendopeptidases ,Tissue Inhibitor of Metalloproteinases ,Sertoli cell ,Stimulation, Chemical ,GRANULOSA-CELLS ,medicine.anatomical_structure ,MESSENGER-RNA ,Proto-Oncogene Proteins c-fos ,RESPONSIVE ELEMENT ,MATRIX METALLOPROTEINASE ,endocrine system ,medicine.medical_specialty ,Molecular Sequence Data ,Cycloheximide ,Biology ,APE LEUKEMIA-VIRUS, PHORBOL ESTER, RESPONSIVE ELEMENT, MATRIX METALLOPROTEINASE, GENE-TRANSCRIPTION, FOS TRANSCRIPTION, GRANULOSA-CELLS, PROTEIN-KINASE, MESSENGER-RNA, JUN ,Internal medicine ,medicine ,Animals ,RNA, Messenger ,Protein kinase A ,JUN ,Protein kinase C ,Glycoproteins ,Tissue Inhibitor of Metalloproteinase-2 ,Messenger RNA ,Binding Sites ,Sertoli Cells ,Base Sequence ,GENE-TRANSCRIPTION ,PHORBOL ESTER ,Proteins ,PROTEIN-KINASE ,Blotting, Northern ,Molecular biology ,Rats ,chemistry ,Tetradecanoylphorbol Acetate ,Follicle Stimulating Hormone ,Oligonucleotide Probes - Abstract
Primary cultures of prepubertal rat Sertoli cells secrete two major tissue inhibitors of metalloproteinases: TIMP-1 (M(r) 28K) and TIMP-2 (M(r) 21 K). FSH stimulated Sertoli cell TIMP-1 and TIMP-2 activity in a time- and dose-dependent manner and also stimulated TIMP-1 and TIMP-2 protein and messenger RNA levels. These effects were mimicked by the cAMP analog, 8-bromo-cAMP, and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine. The protein kinase C activating phorbol ester phorbol myristate acetate (TPA) stimulated TIMP-1 but not TIMP-2 activity and messenger RNA levels. Cycloheximide and actinomycin-D inhibited basal TIMP-1 and TIMP-2 activity and inhibited the ability of FSH, 8-bromo-cAMP, and TPA to stimulate TIMP activity. The protein kinase A (PKA) inhibitor AMP Rp isomer did not affect basal TIMP-1 and TIMP-2 activity or TPA-stimulated TIMP-1 activity. However, the PKA inhibitor markedly reduced FSH and 3-isobutyl-1-methylxanthine stimulation of TIMP-1 and TIMP-2 activity. FSH, 8-bromo-cAMP, and TPA stimuli induced DNA binding complexes capable of binding to a TIMP-1 AP-1 site consensus sequence oligonucleotide. The AP-1 site binding complex(es) induced by all three treatments reacted with antibodies directed broadly against fos and jun protooncogene families and against the specific family members c-fos, junB, and junD but not c-jun proteins. Constitutive cAMP response element binding activity capable of binding an artificial cAMP response element binding site oligonucleotide was demonstrated in Sertoli cell nuclear extracts. This activity was minimally modulated by FSH, 8-bromo-cAMP, or TPA treatment. In summary, Sertoli cells secrete TIMP-1 and TIMP-2 that can be coordinately up-regulated by FSH through a cAMP, PKA-dependent pathway. a convergence of TPA, FSH, and cAMP mediated signals in prepubertal Sertoli cells may occur with the induction of specific AP-1 site binding complex(es) containing jun and fos proteins. Our data suggest that FSH stimulation of TIMP-2 expression may be regulated independently to that of TIMP-1. We propose that the ability of FSH to stimulate Sertoli cell TIMP activity suggests a central role for this hormone in the control of extracellular matrix turnover during testicular development at the level of metalloproteinase inhibition.
26. The Alternative TrkAIII Splice Variant Targets the Centrosome and Promotes Genetic Instability
- Author
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Farina, Antonietta, Tacconelli, A, Cappabianca, LUCIA ANNAMARIA, Cea, G, Panella, S, Chioda, A, Romanelli, A, Pedone, C, Gulino, A, Mackay, A. R., Mackay, ANDREW REAY, Farina, A. R., Tacconelli, A., Cappabianca, L., Cea, G., Panella, S., Chioda, A., Romanelli, Alessandra, Pedone, Carlo, Gulino, A., and Mackay, A. R.
- Subjects
Genome instability ,Chromosomal Proteins, Non-Histone ,Recombinant Fusion Proteins ,Cell Cycle Proteins ,Protein tyrosine phosphatase ,Protein Serine-Threonine Kinases ,Biology ,Genomic Instability ,Cell Line ,Mice ,Neuroblastoma ,Tubulin ,Endopeptidases ,Animals ,Humans ,Protein Isoforms ,Receptor, trkA ,Molecular Biology ,Separase ,Centrosome ,Calcium-Binding Proteins ,Cell Cycle ,Alternative splicing ,Polo kinase ,Exons ,Intracellular Membranes ,Articles ,Cell Biology ,Cell biology ,Enzyme Activation ,Alternative Splicing ,RNA splicing ,Cancer research ,Rabbits ,Signal transduction ,Signal Transduction - Abstract
The hypoxia-regulated alternative TrkAIII splice variant expressed by human neuroblastomas exhibits oncogenic potential, driven by in-frame exon 6 and 7 alternative splicing, leading to omission of the receptor extracellular immunoglobulin C(1) domain and several N-glycosylation sites. Here, we show that the TrkAIII oncogene promotes genetic instability by interacting with and exhibiting catalytic activity at the centrosome. This function depends upon intracellular TrkAIII accumulation and spontaneous interphase-restricted activation, in cytoplasmic tyrosine kinase (tk) domain orientation, predominantly within structures that closely associate with the fully assembled endoplasmic reticulum intermediate compartment and Golgi network. This facilitates TrkAIII tk-mediated binding of gamma-tubulin, which is regulated by endogenous protein tyrosine phosphatases and geldanamycin-sensitive interaction with Hsp90, paving the way for TrkAIII recruitment to the centrosome. At the centrosome, TrkAIII differentially phosphorylates several centrosome-associated components, increases centrosome interaction with polo kinase 4, and decreases centrosome interaction with separase, the net results of which are centrosome amplification and increased genetic instability. The data characterize TrkAIII as a novel internal membrane-associated centrosome kinase, unveiling an important alternative mechanism to "classical" cell surface oncogenic receptor tk signaling through which stress-regulated alternative TrkAIII splicing influences the oncogenic process.
- Published
- 2009
- Full Text
- View/download PDF
27. The neuroblastoma tumour-suppressor TrkAI and its oncogenic alternative TrkAIII splice variant exhibit geldanamycin-sensitive interactions with Hsp90 in human neuroblastoma cells
- Author
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Carlo Pedone, Lucia Cappabianca, Antonella Chioda, Alberto Gulino, Antonietta R. Farina, Andrew R. Mackay, Antonella Tacconelli, Soccorsa Pensato, Gesilia Cea, Alessandra Romanelli, Farina, A. R., Tacconelli, A., Cappabianca, L., Cea, G., Chioda, A., Romanelli, Alessandra, Pensato, S., Pedone, Carlo, Gulino, A., and Mackay, A. R.
- Subjects
Cancer Research ,Tumor suppressor gene ,Lactams, Macrocyclic ,Tropomyosin receptor kinase A ,Transfection ,Receptor tyrosine kinase ,Hsp90 inhibitor ,chemistry.chemical_compound ,neuroblastoma ,Enzyme Stability ,Genetics ,Benzoquinones ,Tumor Cells, Cultured ,Humans ,Genes, Tumor Suppressor ,HSP90 Heat-Shock Proteins ,Phosphorylation ,Receptor, trkA ,geldanamycin ,Molecular Biology ,Protein Kinase Inhibitors ,biology ,Alternative splicing ,Oncogenes ,trkai ,Geldanamycin ,Hsp90 ,hsp90 ,Isoenzymes ,Alternative Splicing ,Protein Transport ,trkaiii ,chemistry ,Antigens, Surface ,biology.protein ,Cancer research ,Tyrosine ,Tyrosine kinase ,Protein Binding - Abstract
Hsp90 chaperones stabilize many tyrosine kinases including several oncogenes, which are inhibited or induced to degrade by the Hsp90 inhibitor geldanamycin (GA). As a consequence, GA has been developed for future chemotherapeutic use in several tumour types including neuroblastoma (NB). Alternative splicing of the neurotrophin receptor tyrosine kinase TrkA may have a pivotal function in regulating NB behaviour, with reports suggesting that tumour-suppressing signals from TrkA may be converted to oncogenic signals by stress-regulated alternative TrkAIII splicing. Within this context, it is important to know whether Hsp90 interacts with TrkA variants in NB cells and how GA influences this. Here, we report that both TrkAI and TrkAIII are Hsp90 clients in human NB cells. TrkAI exhibits GA-sensitive interaction with Hsp90 required for receptor endoplasmic reticulum export, maturation, cell surface stabilization and ligand-mediated activation, whereas TrkAIII exhibits GA-sensitive interactions with Hsp90 required for spontaneous activity and to a lesser extent stability. We show that GA inhibits proliferation and induces apoptosis of TrkAI expressing NB cells, whereas TrkAIII reduces the sensitivity of NB cells to GA-induced elimination. Our data suggest that GA-sensitive interactions with Hsp90 are critical for both TrkAI tumour suppressor and TrkAIII oncogenic function in NB and that TrkAIII expression exerts a negative impact on GA-induced NB cell eradication, which can be counteracted by a novel TrkAIII-specific peptide nucleic acid inhibitor.
- Published
- 2009
28. Group counseling as methodology to prevent homophobic bullying in an action-research project
- Author
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VALERIO, PAOLO, AMODEO, ANNA LISA, L. Cappabianca, M. Canta, R. De Falco, I. Vasta, Valerio, Paolo, Amodeo, ANNA LISA, Cappabianca, L., Canta, M., De Falco, R., Vastola, I., L., Cappabianca, M., Canta, R., De Falco, and I., Vasta
- Subjects
bullismo omofobico ,prevenzione ,counselling di gruppo - Abstract
This scientific work is represented by an action-research project that has been thought with the aim to prevent the homophobic bullying. This project arises from the necessity to train and to sensitize on this problem teachers, parents and other education agency that bear on the development of sexuality social representation. One of the techniques that has been used on this type of project is group counselling with subjects that belong to the same age. This technique is a short term intervention realized in a group with subject that don’t present any psychopathological problems. Group counselling is articulated in two cycles of three sessions each; sessions take place weekly and last for one hour and fifteen minutes. The discussions are conducted by two psychologists, one of whom has the role of conductor and the other that has a non-participant observer role. At the end of each session the task of the second psychologist is to write down the principal topics that have been carried by during the group session. The aim of these group sessions are to think about the topic of the gender identity and try to deal with the problem of homophobic bullying. In the poster the whole project, with a particular focus on the main topics that have came to light during group sessions will be discussed.
- Published
- 2009
29. Pharyngeal disorders: Diagnosis with combined videofluoroscopy and manometry (videfluoromanometry)
- Author
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Cappabianca, S., Brunese, L., Reginelli, A., Pezzullo, M. G., Gianluca GATTA, Grassi, R., Rotondo, A., S. Cappabianca, L. Brunese, A. Reginelli, M.G. Pezzullo, G. Gatta, R Grassi, A. Rotondo, Cappabianca, S., Brunese, L., Reginelli, A., Pezzullo, M. G., Gatta, G., Grassi, R., and Rotondo, A.
- Subjects
Medicine (all)
30. Molecular Characterization and Inhibition of a Novel Stress-Induced Mitochondrial Protecting Role for Misfolded TrkAIII in Human SH-SY5Y Neuroblastoma Cells.
- Author
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Cappabianca L, Ruggieri M, Sebastiano M, Sbaffone M, Martelli I, Ruggeri P, Di Padova M, Farina AR, and Mackay AR
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- Humans, Cell Line, Tumor, Protein Folding, Signal Transduction drug effects, Stress, Physiological drug effects, Endoplasmic Reticulum Chaperone BiP metabolism, Receptor, trkA metabolism, Neuroblastoma metabolism, Neuroblastoma pathology, Mitochondria metabolism, Mitochondria drug effects
- Abstract
Pediatric neuroblastomas (NBs) are heterogeneous, aggressive, therapy-resistant embryonal tumors that originate from cells of neural crest origin committed to the sympathoadrenal progenitor cell lineage. Stress- and drug-resistance mechanisms drive post-therapeutic relapse and metastatic progression, the characterization and inhibition of which are major goals in improving therapeutic responses. Stress- and drug-resistance mechanisms in NBs include alternative TrkAIII splicing of the neurotrophin receptor tropomyosin-related kinase A ( NTRK1/TrkA ), which correlates with post-therapeutic relapse and advanced-stage metastatic disease. The TrkAIII receptor variant exerts oncogenic activity in NB models by mechanisms that include stress-induced mitochondrial importation and activation. In this study, we characterize novel targetable and non-targetable participants in this pro-survival mechanism in TrkAIII-expressing SH-SY5Y NB cells, using dithiothreitol (DTT) as an activator and a variety of inhibitors by regular and immunoprecipitation Western blotting of purified mitochondria and IncuCyte cytotoxicity assays. We report that stress-induced TrkAIII misfolding initiates this mechanism, resulting in Grp78, Ca
2+ -calmodulin, adenosine ribosylating factor (Arf) and Hsp90-regulated mitochondrial importation. TrkAIII imported into inner mitochondrial membranes is cleaved by Omi/high temperature requirement protein A2 (HtrA2) then activated by a mechanism dependent upon calmodulin kinase II (CaMKII), alpha serine/threonine kinase (Akt), mitochondrial Ca2+ uniporter and reactive oxygen species (ROS), involving inhibitory mitochondrial protein tyrosine phosphatase (PTPase) oxidation, resulting in phosphoinositide 3 kinase (PI3K) activation of mitochondrial Akt, which enhances stress resistance. This novel pro-survival function for misfolded TrkAIII mitigates the cytotoxicity of mitochondrial Ca2+ homeostasis disrupted during integrated stress responses, and is prevented by clinically approved Trk and Akt inhibitors and also by inhibitors of 78kDa glucose regulated protein (Grp78), heat shock protein 90 (Hsp90), Ca2+ -calmodulin and PI3K. This identifies Grp78, Ca2+ -calmodulin, Hsp90, PI3K and Akt as novel targetable participants in this mechanism, in addition to TrkAIII, the inhibition of which has the potential to enhance the stress-induced elimination of TrkAIII-expressing NB cells, with the potential to improve therapeutic outcomes in NBs that exhibit TrkAIII expression and activation.- Published
- 2024
- Full Text
- View/download PDF
31. A Study of Alternative TrkA Splicing Identifies TrkAIII as a Novel Potentially Targetable Participant in PitNET Progression.
- Author
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Sbaffone M, Jaffrain-Rea ML, Cappabianca L, Carbonara F, Gianno F, Feola T, Ruggieri M, Zelli V, Maccarone R, Guadagni S, Clementi M, Arcella A, Esposito V, Carozza G, Martelli I, Farina AR, and Mackay AR
- Abstract
Pituitary neuroendocrine tumors (PitNETs) are generally benign but comprise an aggressive, invasive, therapy-resistant, metastatic subset, underpinning a need for novel therapeutic targets. PitNETs exhibit low mutation rates but are associated with conditions linked to alternative splicing, an alternative oncogene pathway activation mechanism. PitNETs express the neurotrophin receptor TrkA, which exhibits oncogenic alternative TrkAIII splicing in other neuroendocrine tumors. We, therefore, assessed whether TrkAIII splicing represents a potential oncogenic participant in PitNETs. TrkAIII splicing was RT-PCR assessed in 53 PitNETs and TrkA isoform(s) expression and activation were assessed by confocal immunofluorescence. TrkAIII splicing was also compared to HIF1α, HIF2α, SF3B1, SRSF2, U2AF1, and JCPyV large T antigen mRNA expression, Xbp1 splicing, and SF3B1 mutation. TrkAIII splicing was detected in all invasive and most non-invasive PitNETs and was significantly elevated in invasive cases. In PitNET lineages, TrkAIII splicing was significantly elevated in invasive PIT1 PitNETs and high in invasive and non-invasive SF1 and TPIT lineages. Immunoreactivity consistent with TrkAIII activation characterized PitNET expressing TrkAIII mRNA, and invasive Pit1 PitNETs exhibited elevated HIF2α expression. TrkAIII splicing did not associate with SF3B1 mutations, altered SF3B1 , SRSF2 , and U2AF1 or JCPyV large T antigen expression, or Xbp1 splicing. Therefore, TrkAIII splicing is common in PitNETs, is elevated in invasive, especially PIT1 tumors, can result in intracellular TrkAIII activation, and may involve hypoxia. The data support a role for TrkAIII splicing in PitNET pathogenesis and progression and identify TrkAIII as a novel potential target in refractory PitNETs.
- Published
- 2024
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32. The Alternative TrkAIII Splice Variant, a Targetable Oncogenic Participant in Human Cutaneous Malignant Melanoma.
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Cappabianca L, Zelli V, Pellegrini C, Sebastiano M, Maccarone R, Clementi M, Chiominto A, Ruggeri P, Cardelli L, Ruggieri M, Sbaffone M, Fargnoli MC, Guadagni S, Farina AR, and Mackay AR
- Subjects
- Humans, Receptor, trkA genetics, Receptor, trkA metabolism, Neoplasm Recurrence, Local, Alternative Splicing genetics, Melanoma, Cutaneous Malignant, Neuroblastoma genetics, Melanoma genetics
- Abstract
Post-therapeutic relapse, poor survival rates and increasing incidence justify the search for novel therapeutic targets and strategies in cutaneous malignant melanoma (CMM). Within this context, a potential oncogenic role for TrkA in CMM is suggested by reports of NTRK1 amplification, enhanced TrkA expression and intracellular TrkA activation associated with poor prognosis. TrkA, however, exhibits tumour-suppressing properties in melanoma cell lines and has recently been reported not to be associated with CMM progression. To better understand these contradictions, we present the first analysis of potential oncogenic alternative TrkA mRNA splicing, associated with TrkA immunoreactivity, in CMMs, and compare the behaviour of fully spliced TrkA and the alternative TrkAIII splice variant in BRAF(V600E) -mutated A375 melanoma cells. Alternative TrkA splicing in CMMs was associated with unfolded protein response (UPR) activation. Of the several alternative TrkA mRNA splice variants detected, TrkAIII was the only variant with an open reading frame and, therefore, oncogenic potential. TrkAIII expression was more frequent in metastatic CMMs, predominated over fully spliced TrkA mRNA expression in ≈50% and was invariably linked to intracellular phosphorylated TrkA immunoreactivity. Phosphorylated TrkA species resembling TrkAIII were also detected in metastatic CMM extracts. In A375 cells, reductive stress induced UPR activation and promoted TrkAIII expression and, in transient transfectants, promoted TrkAIII and Akt phosphorylation, enhancing resistance to reductive stress-induced death, which was prevented by lestaurtinib and entrectinib. In contrast, fully spliced TrkA was dysfunctional in A375 cells. The data identify fully spliced TrkA dysfunction as a novel mechanism for reducing melanoma suppression, support a causal relationship between reductive stress, UPR activation, alternative TrkAIII splicing and TrkAIII activation and characterise a targetable oncogenic pro-survival role for TrkAIII in CMM.
- Published
- 2023
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33. Doxorubicin-Induced TrkAIII Activation: A Selection Mechanism for Resistant Dormant Neuroblastoma Cells.
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Cappabianca L, Sebastiano M, Ruggieri M, Sbaffone M, Zelli V, Farina AR, and Mackay AR
- Subjects
- Alternative Splicing, Benzamides, Calmodulin, Cell Line, Tumor, Crizotinib therapeutic use, Doxorubicin pharmacology, Doxorubicin therapeutic use, Humans, Indazoles, Oxygen therapeutic use, Proto-Oncogene Proteins c-akt, Ryanodine Receptor Calcium Release Channel, Neuroblastoma drug therapy, Neuroblastoma genetics, Receptor, trkA metabolism
- Abstract
Patients with advanced neuroblastoma (NB) receive multimodal clinical therapy, including the potent anthracycline chemotherapy drug doxorubicin (Dox). The acquisition of Dox resistance, however, is a major barrier to a sustained response and leads to a poor prognosis in advanced disease states, reinforcing the need to identify and inhibit Dox resistance mechanisms. In this context, we report on the identification and inhibition of a novel Dox resistance mechanism. This mechanism is characterized by the Dox-induced activation of the oncogenic TrkAIII alternative splice variant, resulting in increased Dox resistance, and is blocked by lestaurtinib, entrectinib, and crizotinib tyrosine kinase and LY294002 IP3-K inhibitors. Using time lapse live cell imaging, conventional and co-immunoprecipitation Western blots, RT-PCR, and inhibitor studies, we report that the Dox-induced TrkAIII activation correlates with proliferation inhibition and is CDK1- and Ca
2+ -uniporter-independent. It is mediated by ryanodine receptors; involves Ca2+ -dependent interactions between TrkAIII, calmodulin and Hsp90; requires oxygen and oxidation; occurs within assembled ERGICs; and does not occur with fully spliced TrkA. The inhibitory effects of lestaurtinib, entrectinib, crizotinib, and LY294002 on the Dox-induced TrkAIII and Akt phosphorylation and resistance confirm roles for TrkAIII and IP3-K consistent with Dox-induced, TrkAIII-mediated pro-survival IP3K/Akt signaling. This mechanism has the potential to select resistant dormant TrkAIII-expressing NB cells, supporting the use of Trk inhibitors during Dox therapy in TrkAIII-expressing NBs.- Published
- 2022
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34. Temporary Botulinum Immobilization of Residuum Muscles for Facilitation of the Initial Ingrowth of Skin to the Porous Skin and Bone Integrated Pylon in the Technology of Direct Skeletal Attachment: Large Animal Model.
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Bohart Z, Cassidy C, Merrill D, Villani M, Villani R, Cappabianca L, and Pitkin M
- Abstract
Enhancing the technology of bone-anchored limb prosthetics, we present a modified porcine model for developing an infection-free integration between the skin and a percutaneous bone implant. The deeply porous Skin and Bone Integrated Pylon (SBIP) presented an infection-free skin-implant interface both after implantation into the dorsum and after implantation into the residuum after below-knee amputation. However, deep ingrowth of skin into the porous cladding of the SBIP was achieved better in the dorsal procedure, while implantation to the residuum sometimes developed a stoma, probably due to the high mobility of the skin and soft tissues in the pig's thigh. Uncontrolled high skin mobility during the first week after implantation constituted a limitation for the porcine animal model, which we tried to address in the current study. As our previous studies showed that casting of the leg residuum did not sufficiently limit the skin's movement around the implant, we tested a modified protocol of the implantation, which included injection of botulinum toxin into the thigh muscles. During the course of the study, we identified proper botulinum toxin componentry, dosage, and the period after injections to achieve a maximal effect of immobilization of the muscles affecting skin movements. To verify the immobilization, we used kinetic data on the asymmetry of loading during gait with the Strideway System, Tekscan, Inc., Boston, MA, USA. We found that injections in the four muscles of the distal thigh of the left hind leg with MYOBLOC® (rimabotulinumtoxinB; 5,000 units/muscle) were sufficient to provide noticeable immobilization by the fourth week after the procedure. This conclusion was made based on the analysis of the dynamics of asymmetry in vertical ground reactions on the injected (left hind) and uninvolved (right hind) legs during gait over an instrumented walkway., Competing Interests: Conflict of Interest: DM, MV, RV, and LC were employed by company DaVinci Biomedical Research Products. MP was employed by Poly-Orth International. The remaining 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.
- Published
- 2022
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35. Hypoxia-induced alternative splicing: the 11th Hallmark of Cancer.
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Farina AR, Cappabianca L, Sebastiano M, Zelli V, Guadagni S, and Mackay AR
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- Disease Progression, Humans, Neoplasms genetics, Alternative Splicing, Epithelial-Mesenchymal Transition, Hypoxia physiopathology, Neoplasms pathology
- Abstract
Hypoxia-induced alternative splicing is a potent driving force in tumour pathogenesis and progression. In this review, we update currents concepts of hypoxia-induced alternative splicing and how it influences tumour biology. Following brief descriptions of tumour-associated hypoxia and the pre-mRNA splicing process, we review the many ways hypoxia regulates alternative splicing and how hypoxia-induced alternative splicing impacts each individual hallmark of cancer. Hypoxia-induced alternative splicing integrates chemical and cellular tumour microenvironments, underpins continuous adaptation of the tumour cellular microenvironment responsible for metastatic progression and plays clear roles in oncogene activation and autonomous tumour growth, tumor suppressor inactivation, tumour cell immortalization, angiogenesis, tumour cell evasion of programmed cell death and the anti-tumour immune response, a tumour-promoting inflammatory response, adaptive metabolic re-programming, epithelial to mesenchymal transition, invasion and genetic instability, all of which combine to promote metastatic disease. The impressive number of hypoxia-induced alternative spliced protein isoforms that characterize tumour progression, classifies hypoxia-induced alternative splicing as the 11th hallmark of cancer, and offers a fertile source of potential diagnostic/prognostic markers and therapeutic targets.
- Published
- 2020
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36. Discovery, characterization and potential roles of a novel NF-YAx splice variant in human neuroblastoma.
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Cappabianca L, Farina AR, Di Marcotullio L, Infante P, De Simone D, Sebastiano M, and Mackay AR
- Subjects
- Animals, Cell Differentiation genetics, Humans, Mice, Neuroblastoma pathology, RNA Splicing, Transcription, Genetic, Transfection, CCAAT-Binding Factor genetics, Neuroblastoma genetics
- Abstract
Background: Identification of novel cancer-associated splice variants is of potential diagnostic, prognostic and therapeutic importance. NF-Y transcription factor is comprised of NF-YA, NF-YB and NF-YC subunits, binds inverted CCAAT-boxes in ≈70% of gene promoters, regulates > 1000 cancer-associated genes and proteins involved in proliferation, staminality, differentiation, apoptosis, metabolism and is subject to component alternative splicing. RT-PCR evaluation of alternative NF-YA splicing in primary human neuroblastomas (NBs), led to discovery of a novel NF-YAx splice variant, also expressed during mouse embryo development and induced by doxorubicin in NB cells. Here, we report the discovery and characterisation of NF-YAx and discus its potential roles in NB., Methods: NF-YAx cDNA was RT-PCR-cloned from a stage 3 NB (provided by the Italian Association of Haematology and Paediatric Oncology, Genova, IT), sequenced and expressed as a protein using standard methods and compared to known fully-spliced NF-YAl and exon B-skipped NF-YAs isoforms in: EMSAs for capacity to form NF-Y complexes; by co-transfection, co-immunoprecipitation and Western blotting for capacity to bind Sp1; by IF for localisation; in AO/EtBr cell-death and colony formation assays for relative cytotoxicity, and by siRNA knockdown, use of inhibitors and Western blotting for potential mechanisms of action. Stable SH-SY5Y transfectants of all three NF-YA isoforms were also propagated and compared by RT-PCR and Western blotting for differences in cell-death and stem cell (SC)-associated gene expression, in cell-death assays for sensitivity to doxorubicin and in in vitro proliferation, substrate-independent growth and in vivo tumour xenograft assays for differences in growth and tumourigenic capacity., Results: NF-YAx was characterized as a novel variant with NF-YA exons B, D and partial F skipping, detected in 20% of NF-YA positive NBs, was the exclusive isoform in a stage 3 NB, expressed in mouse stage E11.5-14 embryos and induced by doxorubicin in SH-SY5Y NB cells. The NF-YAx protein exhibited nuclear localisation, competed with other isoforms in CCAAT box-binding NF-Y complexes but, in contrast to other isoforms, did not bind Sp1. NF-YAx expression in neural-related progenitor and NB cells repressed Bmi1 expression, induced KIF1Bβ expression and promoted KIF1Bβ-dependent necroptosis but in NB cells also selected tumourigenic, doxorubicin-resistant, CSC-like sub-populations, resistant to NF-YAx cytotoxicity., Conclusions: The discovery of NF-YAx in NBs, its expression in mouse embryos and induction by doxorubicin in NB cells, unveils a novel NF-YA splice mechanism and variant, regulated by and involved in development, genotoxic-stress and NB. NF-YAx substitution of other isoforms in NF-Y complexes and loss of capacity to bind Sp1, characterises this novel isoform as a functional modifier of NF-Y and its promotion of KIF1Bβ-dependent neural-lineage progenitor and NB cell necroptosis, association with doxorubicin-induced necroptosis and expression in mouse embryos coinciding with KIF1Bβ-dependent sympathetic neuroblast-culling, confirm a cytotoxic function and potential role in suppressing NB initiation. On the other hand, the in vitro selection of CSC-like NB subpopulations resistant to NF-YAx cytotoxicity not only helps to explain high-level exclusive NF-YAx expression in a stage 3 NB but also supports a role for NF-YAx in disease progression and identifies a potential doxorubicin-inducible mechanism for post-therapeutic relapse.
- Published
- 2019
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37. A pilot study of alternative TrkAIII splicing in Merkel cell carcinoma: a potential oncogenic mechanism and novel therapeutic target.
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Cappabianca L, Guadagni S, Maccarone R, Sebastiano M, Chiominto A, Farina AR, and Mackay AR
- Subjects
- Adult, Aged, Aged, 80 and over, Carcinoma, Merkel Cell, Female, Humans, Male, Middle Aged, Pilot Projects, Receptor, trkA genetics
- Abstract
Background: Merkel cell carcinomas (MCCs) are rare, aggressive, therapeutically-challenging skin tumours that are increasing in incidence and have poor survival rates. The majority are caused by genomic Merkel cell polyomavirus (MCPyV) integration and MCPyV T-antigen expression. Recently, a potential oncogenic role for the tropomyosin-related tyrosine kinase A receptor (TrkA) has been proposed in MCC. Alternative TrkAIII splicing is a TrkA oncogenic activation mechanism that can be promoted by SV40 large T-antigen, an analogue of MCPyV large T-antigen. In this pilot study, therefore, we have evaluated TrkAIII splicing as a novel potential oncogenic mechanism and therapeutic target in MCPyV positive MCC., Methods: Formalin-fixed paraffin-embedded MCC tissues, consisting of 10 stage IV, 1 stage IIIB, 1 stage IIB, 4 stage IIA and 2 stage I tumours, from patients diagnosed and treated from September 2006 to March, 2019, at the University of L'Aquila, L'Aquila, Italy, were compared to 3 primary basal cell carcinomas (BCCs), 3 primary squamous cell carcinomas (SCCs) and 2 normal skin samples by RT-PCR for MCPyV large T-antigen, small T-antigen, VP-1 expression and alternative TrkAIII splicing and by indirect IF for evidence of intracellular TrkA isoform expression and activation., Results: 9 of 10 Recurrent stage IV MCCs were from patients (P.1-3) treated with surgery plus loco-regional Melphalan chemotherapy and remaining MMCs, including 1 stage IV tumour, were from patients treated with surgery alone (P. 4-11). All MCPyV positive MCCs exhibiting MCPyV large T-antigen expression (17 of 18MCCs, 90%) exhibited alternative TrkAIII mRNA splicing (100%), which was exclusive in a significant number and predominant (> 50%) in all stage IV MCCs and the majority of stage 1-III MCCs. MCCs with higher TrkAIII to 18S rRNA expression ratios also exhibited strong or intermediate immunoreactivity to anti-TrkA antibodies, consistent with cytoplasmic TrkAIII expression and activation. In contrast, the MCPyV negative MCC, BCCs, SCCs and normal skin tissues all exhibited exclusive fully-spliced TrkA mRNA expression, associated with variable immunoreactivity for non-phosphorylated but not phosphorylated TrkA., Conclusions: MCPyV positive MCCs but not MCPyV negative MCC, BCCs and SCCs exhibit predominant alternative TrkAIII splicing, with evidence of intracellular TrkAIII activation. This establishes a new potential MCC subset, unveils a novel potential MCPyV oncogenic mechanism and identifies TrkAIII as a novel potential therapeutic target in MCPyV positive MCC.
- Published
- 2019
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38. The oncogenic neurotrophin receptor tropomyosin-related kinase variant, TrkAIII.
- Author
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Farina AR, Cappabianca L, Ruggeri P, Gneo L, Pellegrini C, Fargnoli MC, and Mackay AR
- Subjects
- Alternative Splicing, Animals, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Biomarkers, Tumor, Cell Transformation, Neoplastic genetics, Cell Transformation, Neoplastic metabolism, Gene Deletion, Gene Expression Regulation, Neoplastic, Genetic Association Studies, Genetic Predisposition to Disease, Genomic Instability, Humans, Neoplasms drug therapy, Neoplasms metabolism, Neoplasms pathology, Oncogene Proteins metabolism, Receptor, trkA metabolism, Signal Transduction, Stress, Physiological, Unfolded Protein Response, Genetic Variation, Neoplasms genetics, Oncogene Proteins genetics, Receptor, trkA genetics
- Abstract
Oncogenes derived from the neurotrophin receptor tropomyosin-related kinase TrkA act as drivers in sub-populations of a wide-range of human cancers. This, combined with a recent report that both adult and childhood cancers driven by novel oncogenic TrkA chimeric-fusions exhibit profound, long-lived therapeutic responses to the Trk inhibitor Larotrectinib, highlights the need to improve clinical detection of TrkA oncogene-driven cancers in order to maximise this novel therapeutic potential. Cancers potentially driven by TrkA oncogenes include a proportion of paediatric neuroblastomas (NBs) that express the alternative TrkA splice variant TrkAIII, which exhibits exon 6, 7 and 9 skipping and oncogenic-activity that depends upon deletion of the extracellular D4 Ig-like domain. In contrast to fully spliced TrkA, which exhibits tumour suppressor activity in NB and associates with good prognosis, TrkAIII associates with advanced stage metastatic disease, post therapeutic relapse and worse prognosis, induces malignant transformation of NIH-3T3 cells and exhibits oncogenic activity in NB models. TrkAIII induction in NB cells is stress-regulated by conditions that mimic hypoxia or perturbate the ER with potential to change TrkA tumour-suppressing signals into oncogenic TrkAIII signals within the stressful tumour microenvironment. In contrast to cell surface TrkA, TrkAIII re-localises to intracellular pre-Golgi membranes, centrosomes and mitochondria, within which it exhibits spontaneous ligand-independent activation, triggering a variety of mechanisms that promote tumorigenicity and malignant behaviour, which impact the majority of cancer hallmarks. In this review, we present updates on TrkAIII detection and association with human malignancies, the multiple ways TrkAIII exerts oncogenic activity and potential therapeutic approaches for TrkAIII expressing cancers, with particular reference to NB.
- Published
- 2018
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39. TrkAIII signals endoplasmic reticulum stress to the mitochondria in neuroblastoma cells, resulting in glycolytic metabolic adaptation.
- Author
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Farina AR, Cappabianca L, Gneo L, Ruggeri P, and Mackay AR
- Abstract
Alternative TrkAIII splicing characterises advanced stage metastatic disease and post-therapeutic relapse in neuroblastoma (NB), and in NB models TrkAIII exhibits oncogenic activity. In this study, we report a novel role for TrkAIII in signaling ER stress to the mitochondria in SH-SY5Y NB cells that results in glycolytic metabolic adaptation. The ER stress-inducing agents DTT, A23187 and thapsigargin activated the ER stress-response in control pcDNA SH-SY5Y and TrkAIII expressing SH-SY5Y cells and in TrkAIII SH-SY5Y cells increased TrkAIII targeting to mitochondria and internalisation into inner-mitochondrial membranes. Within inner-mitochondrial membranes, TrkAIII was subjected to Omi/HtrA2-dependent cleavage to tyrosine phosphorylated 45-48kDa carboxyl terminal active fragments, localised predominantly in tyrosine kinase-domain mitochondrial matrix orientation. This stress-induced activation of mitochondrial TrkAIII was associated with increased ROS production, prevented by the ROS scavenger Resveratrol and underpinned by changes in Ca2+ movement, implicating ROS/Ca2+ interplay in overcoming the mitochondrial TrkAIII activation threshold. Stress-induced, cleavage-activation of mitochondrial TrkAIII resulted in mitochondrial PDHK1 tyrosine phosphorylation, leading to glycolytic metabolic adaptation. This novel mitochondrial role for TrkAIII provides a potential self-perpetuating, drug reversible way through which tumour microenvironmental stress may maintain the metastasis promoting "Warburg effect" in TrkAIII expressing NBs., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of Interest
- Published
- 2017
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40. TRAIL induces pro-apoptotic crosstalk between the TRAIL-receptor signaling pathway and TrkAIII in SH-SY5Y cells, unveiling a potential therapeutic "Achilles heel" for the TrkAIII oncoprotein in neuroblastoma.
- Author
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Gneo L, Ruggeri P, Cappabianca L, Farina AR, Di Ianni N, and Mackay AR
- Subjects
- Apoptosis Regulatory Proteins genetics, Apoptosis Regulatory Proteins metabolism, Cell Line, Tumor, Dose-Response Relationship, Drug, Humans, Neuroblastoma genetics, Neuroblastoma metabolism, Neuroblastoma pathology, Phosphorylation, Protein Tyrosine Phosphatases, Non-Receptor genetics, Protein Tyrosine Phosphatases, Non-Receptor metabolism, RNA Interference, Receptor, trkA genetics, Receptors, TNF-Related Apoptosis-Inducing Ligand genetics, Receptors, TNF-Related Apoptosis-Inducing Ligand metabolism, Time Factors, Transfection, src-Family Kinases genetics, src-Family Kinases metabolism, Antineoplastic Agents pharmacology, Apoptosis drug effects, Drug Resistance, Neoplasm genetics, Neuroblastoma drug therapy, Receptor Cross-Talk, Receptor, trkA metabolism, Receptors, TNF-Related Apoptosis-Inducing Ligand agonists, Signal Transduction drug effects, TNF-Related Apoptosis-Inducing Ligand pharmacology
- Abstract
TrkAIII expression in neuroblastoma (NB) associates with advanced stage disease, worse prognosis, post therapeutic relapse, and in NB models TrkAIII exhibits oncogenic activity and promotes chemotherapeutic-resistance. Here, we report a potential therapeutic "Achilles heel" for the TrkAIII oncoprotein in a SH-SY5Y NB model that is characterised by one-way TRAIL-induced, pro-apoptotic crosstalk between the TRAIL receptor signaling pathway and TrkAIII that results in the delayed induction of apoptosis. In TrkAIII SH-SY5Y cells, blocked in the intrinsic apoptosis pathway by elevated constitutive Bcl-2, Bcl-xL and Mcl-1 expression, TRAIL induced delayed caspase-dependent apoptosis via the extrinsic pathway and completely abrogated tumourigenic capacity in vitro. This effect was initiated by TRAIL-induced SHP-dependent c-Src activation, the induction of TrkAIII/SHP-1/c-Src complexing leading to SHP-mediated TrkAIII de-phosphorylation, subsequent induction of complexing between de-phosphorylated TrkAIII and cFLIP associated with a time-dependent increase the caspase-8 to cFLIP ratio at activated death receptors, resulting in delayed caspase cleavage and caspase-dependent apoptosis. We also confirm rate-limiting roles for c-FLIP and Mcl-1 in regulating the sensitivity of TrkAIII SH-SY5Y cells to TRAIL-induced apoptosis via the extrinsic and intrinsic pathways, respectively. Our study unveils a novel mechanism for the TRAIL-induced apoptosis of TrkAIII expressing NB cells that depends upon SHP/Src-mediated crosstalk between the TRAIL-receptor signaling pathway and TrkAIII, and supports a novel potential pro-apoptotic therapeutic use for TRAIL in TrkAIII expressing NB.
- Published
- 2016
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41. Retrograde TrkAIII transport from ERGIC to ER: a re-localisation mechanism for oncogenic activity.
- Author
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Farina AR, Cappabianca L, Ruggeri P, Gneo L, Maccarone R, and Mackay AR
- Subjects
- Alternative Splicing genetics, Carcinogenesis genetics, Cell Line, Tumor, Glycosylation drug effects, Humans, Mutation genetics, Neuroblastoma drug therapy, Neuroblastoma genetics, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Protein Structure, Tertiary genetics, Protein Transport drug effects, Protein Transport genetics, Receptor, trkA genetics, Signal Transduction drug effects, Signal Transduction genetics, Endoplasmic Reticulum metabolism, Golgi Apparatus metabolism, Neuroblastoma metabolism, Oncogene Protein v-akt metabolism, Receptor, trkA metabolism
- Abstract
In human SH-SY5Y neuroblastoma (NB) cells, nascent immature N-glycosylated 110kDa TrkA moves rapidly from the endoplasmic reticulum (ER) to the Golgi Network (GN), where it matures into the 140kDa receptor prior to being transported to the cell surface, creating GN and cell surface pools of inactive receptor maintained below the spontaneous activation threshold by a full compliment of inhibitory domains and endogenous PTPases. In contrast, the oncogenic alternative TrkAIII splice variant is not expressed at the cell surface but re-localises to intracellular membranes, within which it exhibits spontaneous ERGIC/COPI-associated activation and oncogenic Akt signalling. In this study, we characterise the mechanism responsible for TrkAIII re-localisation. Spontaneous TrkAIII activation, facilitated by D4 IG-like domain and N-glycosylation site omission, increases spontaneous activation potential by altering intracellular trafficking, inhibiting cell surface expression and eliminating an important inhibitory domain. TrkAIII, spontaneously activated within the permissive ERGIC/COPI compartment, rather than moving in an anterograde direction to the GN exhibits retrograde transport back to the ER, where it is inactivated. This sets-up self-perpetuating TrkAIII re-cycling between the ERGIC and ER, that ensures continual accumulation above the spontaneous activation threshold of the ERGIC/COPI compartment. This is reversed by TrkA tyrosine kinase inhibitors, which promote anterograde transport of inactivated TrkAIII to the GN, resulting in GN-associated TrkAIII maturation to a 120kDa species that is degraded at the proteasome.
- Published
- 2015
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42. The TrkAIII oncoprotein inhibits mitochondrial free radical ROS-induced death of SH-SY5Y neuroblastoma cells by augmenting SOD2 expression and activity at the mitochondria, within the context of a tumour stem cell-like phenotype.
- Author
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Ruggeri P, Farina AR, Di Ianni N, Cappabianca L, Ragone M, Ianni G, Gulino A, and Mackay AR
- Subjects
- Carcinogenesis drug effects, Cell Death drug effects, Cell Differentiation drug effects, Cell Line, Tumor, Cell Survival drug effects, Disease Progression, Drug Resistance, Neoplasm, Gene Knockdown Techniques, Humans, Mitochondria drug effects, NF-kappa B metabolism, Neoplastic Stem Cells drug effects, Oncogene Proteins antagonists & inhibitors, Oncogene Proteins metabolism, Phenotype, Phosphotransferases (Alcohol Group Acceptor) metabolism, Protein Isoforms antagonists & inhibitors, Protein Isoforms metabolism, Protein Kinase Inhibitors pharmacology, Receptor, trkA antagonists & inhibitors, Spheroids, Cellular drug effects, Spheroids, Cellular pathology, Superoxide Dismutase deficiency, Gene Expression Regulation, Neoplastic drug effects, Mitochondria metabolism, Neoplastic Stem Cells pathology, Neuroblastoma pathology, Reactive Oxygen Species metabolism, Receptor, trkA metabolism, Superoxide Dismutase genetics
- Abstract
The developmental and stress-regulated alternative TrkAIII splice variant of the NGF receptor TrkA is expressed by advanced stage human neuroblastomas (NBs), correlates with worse outcome in high TrkA expressing unfavourable tumours and exhibits oncogenic activity in NB models. In the present study, we report that constitutive TrkAIII expression in human SH-SY5Y NB cells inhibits Rotenone, Paraquat and LY83583-induced mitochondrial free radical reactive oxygen species (ROS)-mediated death by stimulating SOD2 expression, increasing mitochondrial SOD2 activity and attenuating mitochondrial free radical ROS production, in association with increased mitochondrial capacity to produce H2O2, within the context of a more tumour stem cell-like phenotype. This effect can be reversed by the specific TrkA tyrosine kinase inhibitor GW441756, by the multi-kinase TrkA inhibitors K252a, CEP-701 and Gö6976, which inhibit SOD2 expression, and by siRNA knockdown of SOD2 expression, which restores the sensitivity of TrkAIII expressing SH-SY5Y cells to Rotenone, Paraquat and LY83583-induced mitochondrial free radical ROS production and ROS-mediated death. The data implicate the novel TrkAIII/SOD2 axis in promoting NB resistance to mitochondrial free radical-mediated death and staminality, and suggest that the combined use of TrkAIII and/or SOD2 inhibitors together with agents that induce mitochondrial free radical ROS-mediated death could provide a therapeutic advantage that may also target the stem cell niche in high TrkA expressing unfavourable NB.
- Published
- 2014
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43. TrkAIII promotes microtubule nucleation and assembly at the centrosome in SH-SY5Y neuroblastoma cells, contributing to an undifferentiated anaplastic phenotype.
- Author
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Farina AR, Di Ianni N, Cappabianca L, Ruggeri P, Ragone M, Ianni G, Gulino A, and Mackay AR
- Subjects
- Cell Line, Tumor, Centrosome pathology, Gene Expression Regulation, Neoplastic, Humans, Microtubules metabolism, Microtubules pathology, Neoplasm Staging, Neuroblastoma pathology, Phosphorylation, Protein Binding, Receptor, trkA metabolism, Signal Transduction genetics, Alternative Splicing genetics, Neuroblastoma genetics, Receptor, trkA genetics, Tubulin metabolism
- Abstract
The alternative TrkAIII splice variant is expressed by advanced stage human neuroblastomas (NBs) and exhibits oncogenic activity in NB models. In the present study, employing stable transfected cell lines and assays of indirect immunofluorescence, immunoprecipitation, Western blotting, microtubule regrowth, tubulin kinase, and tubulin polymerisation, we report that TrkAIII binds α -tubulin and promotes MT nucleation and assembly at the centrosome. This effect depends upon spontaneous TrkAIII activity, TrkAIII localisation to the centrosome and pericentrosomal area, and the capacity of TrkAIII to bind, phosphorylate, and polymerise tubulin. We propose that this novel role for TrkAIII contributes to MT involvement in the promotion and maintenance of an undifferentiated anaplastic NB cell morphology by restricting and augmenting MT nucleation and assembly at the centrosomal MTOC.
- Published
- 2013
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44. Constitutive autotaxin transcription by Nmyc-amplified and non-amplified neuroblastoma cells is regulated by a novel AP-1 and SP-mediated mechanism and abrogated by curcumin.
- Author
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Farina AR, Cappabianca L, Ruggeri P, Di Ianni N, Ragone M, Merolle S, Sano K, Stracke ML, Horowitz JM, Gulino A, and Mackay AR
- Subjects
- Cell Line, Tumor, Cyclic AMP genetics, Gene Deletion, Genes, Reporter genetics, Humans, Phosphoric Diester Hydrolases deficiency, Response Elements drug effects, Response Elements genetics, Transcriptional Activation drug effects, Curcumin pharmacology, Neuroblastoma pathology, Phosphoric Diester Hydrolases genetics, Proto-Oncogene Proteins c-myc metabolism, Sp Transcription Factors metabolism, Transcription Factor AP-1 metabolism, Transcription, Genetic drug effects
- Abstract
The motility, angiogenesis and metastasis-stimulating factor Autotaxin (Atx), over expressed by human neuroblastomas (NB), is constitutively expressed by human Nmyc-amplified SK-N-BE and non-Nmyc-amplified SH-SY5Y NB cells. Here, we characterise a novel Atx transcriptional mechanism, utilised by both cell lines, that is restricted to the first 285bp of the Atx promoter and involves AP-1 and SP transcription factors, acting through a CRE/AP-1-like element at position -142 to -149 and a GAbox at position -227 to -235 relative to the Atx translational start site. This novel transcriptional mechanism can be inhibited by internally initiated SP-3 and the natural phenol curcumin., (Copyright © 2012 Federation of European Biochemical Societies. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
45. Alendronate promotes plasmin-mediated MMP-9 inactivation by exposing cryptic plasmin degradation sites within the MMP-9 catalytic domain.
- Author
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Farina AR, Cappabianca L, Di Ianni N, Ruggeri P, Ragone M, Merolle S, Gulino A, and Mackay AR
- Subjects
- Catalysis, Catalytic Domain, Cations, Cell Line, Tumor, Chelating Agents pharmacology, Dose-Response Relationship, Drug, Edetic Acid chemistry, Hemopexin chemistry, Humans, Plasminogen chemistry, Protein Structure, Tertiary, Recombinant Proteins chemistry, Alendronate pharmacology, Fibrinolysin metabolism, Matrix Metalloproteinase 9 metabolism
- Abstract
Irreversible MMP-9 inhibition is considered a significant therapeutic goal in inflammatory, vascular and tumour pathology. We report that divalent cation chelators Alendronate and EDTA not only directly inhibited MMP-9 but also promoted irreversible plasmin-mediated MMP-9 inactivation by exposing cryptic plasmin-degradation sites within the MMP-9 catalytic-domain and producing an inhibitory hemopexin-domain fragment. This effect was also observed using MDA-MB-231 breast cancer cells, which activated exogenous plasminogen to degrade endogenous proMMP-9 in the presence of Alendronate or EDTA. Degradation-mediated inactivation of proMMP-9 occurred in the absence of transient activation, attesting to the incapacity of plasmin to directly activate proMMP-9 and direct MMP-9 inhibition by Alendronate and EDTA. Our study provides a novel rational for therapeutic Alendronate use in MMP-9-dependent pathology characterised by plasminogen activation., (Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
46. Thioredoxin stimulates MMP-9 expression, de-regulates the MMP-9/TIMP-1 equilibrium and promotes MMP-9 dependent invasion in human MDA-MB-231 breast cancer cells.
- Author
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Farina AR, Cappabianca L, DeSantis G, Di Ianni N, Ruggeri P, Ragone M, Merolle S, Tonissen KF, Gulino A, and Mackay AR
- Subjects
- Breast Neoplasms genetics, Breast Neoplasms pathology, Female, Histone Deacetylases genetics, Histone Deacetylases metabolism, Humans, Matrix Metalloproteinase 9 genetics, NF-kappa B p50 Subunit genetics, NF-kappa B p50 Subunit metabolism, Neoplasm Invasiveness, Neoplasm Proteins genetics, Response Elements genetics, Thioredoxins genetics, Tissue Inhibitor of Metalloproteinase-1 genetics, Transcription Factor RelA genetics, Transcription Factor RelA metabolism, Transcription, Genetic genetics, Breast Neoplasms metabolism, Gene Expression Regulation, Enzymologic, Gene Expression Regulation, Neoplastic, Matrix Metalloproteinase 9 biosynthesis, Neoplasm Proteins metabolism, Thioredoxins metabolism, Tissue Inhibitor of Metalloproteinase-1 metabolism
- Abstract
Increased expression of thioredoxin (Trx)-1 and matrix metalloproteinase (MMP)-9 associates with malignant breast cancer progression. Here, we describe a functional relationship between Trx-1 and MMP-9 in promoting MDA-MB-231 breast cancer cell invasive behaviour. Trx-1 overexpression stimulated MMP-9 expression, de-regulated the MMP-9/TIMP-1 equilibrium and augmented MMP-9 involvement in a more invasive phenotype. Trx-1 augmented MMP-9 transcription through NF-κB, AP-1 and SP1 elements; stimulated p50/p65 NF-κB activity and recruitment to the MMP-9 promoter; and facilitated MMP-9 promoter-accessibility to NF-κB by preventing HDAC recruitment and maintaining MMP-9 promoter histone acetylation. Our data provide a functional basis for Trx-1 and MMP-9 association in malignant breast cancer and identify Trx-1 and NF-κB as potentially druggable targets for reducing MMP-9 involvement in malignant behaviour., (Copyright © 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
47. TrkAIII expression in the thymus.
- Author
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Tacconelli A, Farina AR, Cappabianca L, Cea G, Panella S, Chioda A, Gallo R, Cinque B, Sferra R, Vetuschi A, Campese AF, Screpanti I, Gulino A, and Mackay AR
- Subjects
- Animals, Animals, Newborn, Antigens, CD metabolism, Cells, Cultured, Embryo, Mammalian, Epithelial Cells physiology, Flow Cytometry methods, Gene Expression Regulation, Neoplastic, Humans, Immunohistochemistry methods, Jurkat Cells, Mice, Neuroblastoma, Thymus Extracts metabolism, Receptor, trkA genetics, Receptor, trkA metabolism, Thymus Gland cytology, Thymus Gland metabolism
- Abstract
The alternative TrkAIII splice variant is expressed by murine and human thymus. Alternative TrkAIII splicing predominates in postembryonic day E13 (E17 and E18), postnatal murine (3 week and 3 month) and human thymuses, with TrkAIII mRNA expressed by selected thymocyte subsets and thymic epithelial cells (TECs) and a 100 kDa immunoprecipitable TrkAIII-like protein detected in purified thymocyte and whole thymus extracts. FACS and immunohistochemical analysis indicate a non-cell surface localisation for the TrkAIII-like protein in cortical CD4+/CD8+ double positive and, to a lesser extent, single positive thymocyte subsets at the cortex/medulla boundary and in Hassle's corpuscles, reticular epithelial and dendritic cells of the thymic medulla. TrkA(I/II) expression, on the other hand, predominates in sub-capsular regions of the thymus. TrkAIII-like immunoreactivity at the cortex/medulla boundary associates with regions of thymocyte proliferation and not apoptosis. A potential role for thymic hypoxia in thymocyte alternative TrkAIII splicing is supported by reversal to TrkAI splicing by normoxic but not hypoxic culture and induction of Jurkat T cell alternative TrkAIII splicing by the hypoxia mimic CoCl2. In contrast, TEC expression of TrkAIII predominates in both normoxic and hypoxic culture conditions. The data support a potential role for TrkAIII in thymic development and function, of particular relevance to intermediate stage CD4+/CD8+ thymocyte subsets and TECs, which potentially reflects a reversible thymocyte and more permanent TEC adaptation to thymic environment. Since intracellular TrkAIII neither binds nor responds to NGF and can impede regular NGF/TrkA signalling (Tacconelli et al., Cancer Cell, 2004), its expression would be expected to provide an alternative and/or impediment to regular NGF/TrkA signalling within the developing and developed thymus of potential functional importance.
- Published
- 2007
- Full Text
- View/download PDF
48. Alternative TrkAIII splicing: a potential regulated tumor-promoting switch and therapeutic target in neuroblastoma.
- Author
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Tacconelli A, Farina AR, Cappabianca L, Gulino A, and Mackay AR
- Subjects
- Animals, Gene Expression Regulation, Neoplastic, Humans, Receptor, trkA antagonists & inhibitors, Receptor, trkA metabolism, Signal Transduction, Alternative Splicing genetics, Neuroblastoma genetics, Neuroblastoma metabolism, Neuroblastoma pathology, Receptor, trkA genetics
- Abstract
An association between elevated tyrosine kinase receptor (Trk)-A expression and better prognosis; the absence of mutation-activated TrkA oncogenes; the induction of apoptosis, growth arrest, morphological differentiation and inhibition of xenograft growth; and angiogenesis by TrkA gene transduction, provide the basis for the current concept of an exclusively tumor-suppressor role for TrkA in the aggressive pediatric tumor, neuroblastoma. This concept, however, has recently been challenged by the discovery of a novel hypoxia-regulated alternative TrkAIII splice variant, initial data for which suggest predominant expression in advanced-stage neuroblastoma. TrkAIII exhibits neuroblastoma xenograft tumor-promoting activity associated with the induction of a more angiogenic and stress-resistant neuroblastoma phenotype and antagonises nerve growth factor/TrkAI antioncogenic signaling. In this short review, the authors integrate this novel information into a modified concept that places alternative TrkA splicing as a potential pivotal regulator of neuroblastoma behavior and identifies the TrkAIII alternative splice variant as a potential biomarker of patient prognosis and novel therapeutic target.
- Published
- 2005
- Full Text
- View/download PDF
49. TrkAIII. A novel hypoxia-regulated alternative TrkA splice variant of potential physiological and pathological importance.
- Author
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Tacconelli A, Farina AR, Cappabianca L, Gulino A, and Mackay AR
- Subjects
- Cell Differentiation genetics, Cell Line, Tumor, Cell Proliferation, DNA, Neoplasm genetics, Gene Expression Regulation, Neoplastic, Humans, Neoplastic Stem Cells pathology, Neoplastic Stem Cells physiology, Neural Crest cytology, Neural Crest physiology, Neuroblastoma genetics, Neuroblastoma pathology, Neuroblastoma physiopathology, Signal Transduction genetics, Alternative Splicing, Cell Hypoxia, Genetic Variation, Receptor, trkA genetics, Receptor, trkA physiology
- Abstract
Nerve growth factor receptor TrkA is critical for development and maturation of central and peripheral nervous systems, regulating proliferation, differentiation and apoptosis. In cancer, TrkA frequently exhibits suppressor activity in nonmutated form and oncogenic activity upon mutation. Our identification of a novel hypoxia-regulated alternative TrkAIII splice variant, expressed by neural crest-derived neuroblastic tumors, that exhibits neuroblastoma tumor promoting activity, adds significantly to our understanding of potential TrkA involvement in cancer. Our observation that hypoxia, which characterizes the tumor micro-environment, stimulates alternative TrkAIII splicing, provides a way by which TrkA tumor suppressing signals may convert to tumor promoting signals during progression and is consistent with conservation and pathological subversion by neural crest-derived neuroblastic tumors of a mechanism of potential physiological importance to normal neural stem/neural crest progenitors.
- Published
- 2005
- Full Text
- View/download PDF
50. TrkA alternative splicing: a regulated tumor-promoting switch in human neuroblastoma.
- Author
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Tacconelli A, Farina AR, Cappabianca L, Desantis G, Tessitore A, Vetuschi A, Sferra R, Rucci N, Argenti B, Screpanti I, Gulino A, and Mackay AR
- Subjects
- Adaptor Proteins, Signal Transducing metabolism, Amino Acid Sequence, Animals, Apoptosis drug effects, Base Sequence, Cell Line, Cloning, Molecular, Doxorubicin pharmacology, Gene Expression Regulation, Neoplastic, Humans, Hypoxia genetics, Mice, Molecular Sequence Data, NF-kappa B metabolism, Neovascularization, Pathologic, Nerve Growth Factor metabolism, Nerve Growth Factor pharmacology, Neuroblastoma blood supply, Neuroblastoma genetics, Phosphatidylinositol 3-Kinases metabolism, Phospholipase C gamma, Protein Binding, Receptor, trkA antagonists & inhibitors, Receptor, trkA chemistry, Shc Signaling Adaptor Proteins, Signal Transduction, Src Homology 2 Domain-Containing, Transforming Protein 1, Type C Phospholipases metabolism, Alternative Splicing genetics, Neuroblastoma metabolism, Neuroblastoma pathology, Receptor, trkA genetics, Receptor, trkA metabolism
- Abstract
We identify a novel alternative TrkA splice variant, TrkAIII, with deletion of exons 6, 7, and 9 and functional extracellular IG-C1 and N-glycosylation domains, that exhibits expression restricted to undifferentiated early neural progenitors, human neuroblastomas (NBs), and a subset of other neural crest-derived tumors. This NGF-unresponsive isoform is oncogenic in NIH3T3 cells and promotes tumorigenic NB cell behavior in vitro and in vivo (cell survival, xenograft growth, angiogenesis) resulting from spontaneous tyrosine kinase activity and IP3K/Akt/NF-kappaB but not Ras/MAPK signaling. TrkAIII antagonizes NGF/TrkAI signaling, which is responsible for NB growth arrest and differentiation through Ras/MAPK, and its expression is promoted by hypoxia at the expense of NGF-responsive receptors, providing a mechanism for converting NGF/TrkA/Ras/MAPK antioncogenic signals to TrkAIII/IP3K/Akt/NF-kappaB tumor-promoting signals during tumor progression.
- Published
- 2004
- Full Text
- View/download PDF
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