title: Heat shock protein 90 inhibition depletes TrkA levels and signaling in human acute leukemia cells.
abstract: Nerve growth factor (NGF) induces autophosphorylation and downstream progrowth and prosurvival signaling from the receptor tyrosine kinase TrkA. Overexpression or activating mutation of TrkA has been described in human acute myeloid leukemia cells. In the present study, we show the chaperone association of TrkA with heat shock protein 90 (hsp90) and the inhibitory effect of the hsp90 inhibitor, 17-DMAG, on TrkA levels and signaling in cultured and primary myeloid leukemia cells. Treatment with 17-DMAG disrupted the binding of TrkA with hsp90 and the cochaperone cdc37, resulting in polyubiquitylation, proteasomal degradation, and depletion of TrkA. Exposure to 17-DMAG inhibited NGF-induced p-TrkA, p-AKT, and p-ERK1/2 levels, as well as induced apoptosis of K562, 32D cells with ectopic expression of wild-type TrkA or the constitutively active mutant Delta TrkA, and of primary myeloid leukemia cells. Additionally, 17-DMAG treatment inhibited NGF-induced neurite formation in the rat pheochromocytoma PC-12 cells. Cotreatment with 17-DMAG and K-252a, an inhibitor of TrkA-mediated signaling, induced synergistic loss of viability of cultured and primary myeloid leukemia cells. These findings show that TrkA is an hsp90 client protein, and inhibition of hsp90 depletes TrkA and its progrowth and prosurvival signaling in myeloid leukemia cells. These findings also support further evaluation of the combined activity of an hsp90 inhibitor and TrkA antagonist against myeloid leukemia cells.
authors.affiliation: Medical College of Georgia Cancer Center, 1120 15th Street CN 2133, Augusta, GA 30912, USA.
authors.forename: Rekha
authors.initials: R
authors.lastname: Rao
authors.forename: Srilatha
authors.initials: S
authors.lastname: Nalluri
authors.forename: Warren
authors.initials: W
authors.lastname: Fiskus
authors.forename: Ramesh
authors.initials: R
authors.lastname: Balusu
authors.forename: Atul
authors.initials: A
authors.lastname: Joshi
authors.forename: Uma
authors.initials: U
authors.lastname: Mudunuru
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Buckley
authors.forename: Kelly
authors.initials: K
authors.lastname: Robbins
authors.forename: Celalettin
authors.initials: C
authors.lastname: Ustun
authors.forename: Gary W
authors.initials: GW
authors.lastname: Reuther
authors.forename: Kapil N
authors.initials: KN
authors.lastname: Bhalla
mesh_terms: D000818:Animals; D016227:Benzoquinones / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology; D002227:Carbazoles / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D026121:Indole Alkaloids / Q000494:pharmacology; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism* / Q000473:pathology*; D020932:Nerve Growth Factor / Q000494:pharmacology; D016716:PC12 Cells; D010766:Phosphorylation / Q000187:drug effects; D025821:Polyubiquitin / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D051381:Rats; D020917:Receptor, trkA / Q000378:metabolism*; D015398:Signal Transduction; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects; D054875:Ubiquitination / Q000187:drug effects
chemical_list: D016227:Benzoquinones; D002227:Carbazoles; D018841:HSP90 Heat-Shock Proteins; D026121:Indole Alkaloids; D047029:Lactams, Macrocyclic; C448659:17-(dimethylaminoethylamino)-17-demethoxygeldanamycin; D025821:Polyubiquitin; D020932:Nerve Growth Factor; C049985:staurosporine aglycone; D020917:Receptor, trkA; D046988:Proteasome Endopeptidase Complex