Hsp90 mediates insulin-like growth factor 1 and interleukin-1beta signaling in an age-dependent manner in equine articular chondrocytes.
OBJECTIVE
Many metabolic processes in chondrocytes thought to contribute to age-related changes in the extracellular matrix are influenced by known roles of Hsp90. Age-related decreases in the level of Hsp90 have been documented in numerous cell types and could contribute to cartilage degeneration. The aim of this study was to investigate the roles of age and Hsp90 in insulin-like growth factor 1 (IGF-1) and interleukin-1beta (IL-1beta) signaling in chondrocytes.


METHODS
Levels of Hsp90 messenger RNA (mRNA) and protein, with respect to age, were determined by quantitative real-time polymerase chain reaction (PCR) and Western blot analysis, respectively. The Hsp90 inhibitor geldanamycin (50 nM, 100 nM, or 500 nM) was used to assess age-related responses to Hsp90 with concurrent IGF-1 or IL-1beta stimulation of chondrocytes. Quantitative real-time PCR was used to measure COL2A1 and matrix metalloproteinase 13 (MMP13) gene expression; Western blot analysis was performed to determine the phosphorylation status of p42/44 and Akt/protein kinase B.


RESULTS
The effects of Hsp90 inhibition with geldanamycin were concentration dependent. Inhibition of Hsp90 with 100 nM or 500 nM geldanamycin blocked IGF-1-induced cell proliferation, Akt and p42/44 activation, and COL2A1 expression. Basal and IL-1beta-induced up-regulation of MMP13 mRNA was blocked by all concentrations of geldanamycin tested. Gain-of-function assays with Hsp90 resulted in increased expression of MMP13 mRNA.


CONCLUSIONS
These results suggest that Hsp90 is involved in opposing signaling pathways of cartilage homeostasis, and that catabolic responses are more sensitive to Hsp90 inhibition than are anabolic responses. Further studies are needed to determine the role of Hsp90 inhibition in osteoarthritis in order to assess its potential as a therapeutic target.
Boehm
Amber K
AK
Cornell University, Veterinary Medical Center, Ithaca, New York 14853, USA.
Seth
Mayank
M
Mayr
Karen G
KG
Fortier
Lisa A
LA
D000375:Aging; D000818:Animals; D016227:Benzoquinones / Q000494:pharmacology; D002358:Cartilage, Articular / Q000254:growth & development / Q000502:physiology*; D019902:Chondrocytes / Q000502:physiology*; D024043:Collagen Type II; D018507:Gene Expression Regulation, Developmental; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics*; D006736:Horses; D007334:Insulin-Like Growth Factor I / Q000502:physiology*; D053583:Interleukin-1beta / Q000502:physiology*; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D020782:Matrix Metalloproteinases / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction
D016227:Benzoquinones; D024043:Collagen Type II; D018841:HSP90 Heat-Shock Proteins; D053583:Interleukin-1beta; D047029:Lactams, Macrocyclic; D012333:RNA, Messenger; D007334:Insulin-Like Growth Factor I; D020782:Matrix Metalloproteinases; C001277:geldanamycin