The neuroblastoma tumour-suppressor TrkAI and its oncogenic alternative TrkAIII splice variant exhibit geldanamycin-sensitive interactions with Hsp90 in human neuroblastoma cells.
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.
Farina
A R
AR
Department of Experimental Medicine, University of L'Aquila, L'Aquila, Italy.
Tacconelli
A
A
Cappabianca
L
L
Cea
G
G
Chioda
A
A
Romanelli
A
A
Pensato
S
S
Pedone
C
C
Gulino
A
A
Mackay
A R
AR
D017398:Alternative Splicing; D000954:Antigens, Surface / Q000378:metabolism; D016227:Benzoquinones / Q000494:pharmacology*; D004795:Enzyme Stability / Q000187:drug effects; D016147:Genes, Tumor Suppressor / Q000187:drug effects / Q000502:physiology; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism / Q000502:physiology; D047029:Lactams, Macrocyclic / Q000494:pharmacology*; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D009857:Oncogenes / Q000187:drug effects / Q000502:physiology; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D020917:Receptor, trkA / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism
D000954:Antigens, Surface; D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D007527:Isoenzymes; D047029:Lactams, Macrocyclic; D047428:Protein Kinase Inhibitors; D014443:Tyrosine; D020917:Receptor, trkA; C001277:geldanamycin