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BMC Musculoskeletal Disorders
Published in: BMC Musculoskeletal Disorders 1/2022

01-12-2022 | Shock | Research

Upregulation of TGF-β-induced HSP27 by HSP90 inhibitors in osteoblasts

Authors: Gen Kuroyanagi, Haruhiko Tokuda, Kazuhiko Fujita, Tetsu Kawabata, Go Sakai, Woo Kim, Tomoyuki Hioki, Junko Tachi, Rie Matsushima-Nishiwaki, Takanobu Otsuka, Hiroki Iida, Osamu Kozawa

Published in: BMC Musculoskeletal Disorders | Issue 1/2022

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Abstract

Background

Heat shock protein (HSP) 90 functions as a molecular chaperone and is constitutively expressed and induced in response to stress in many cell types. We have previously demonstrated that transforming growth factor-β (TGF-β), the most abundant cytokine in bone cells, induces the expression of HSP27 through Smad2, p44/p42 mitogen-activated protein kinase (MAPK), p38 MAPK, and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) in mouse osteoblastic MC3T3-E1 cells. This study investigated the effects of HSP90 on the TGF-β-induced HSP27 expression and the underlying mechanism in mouse osteoblastic MC3T3-E1 cells.

Methods

Clonal osteoblastic MC3T3-E1 cells were treated with the HSP90 inhibitors and then stimulated with TGF-β. HSP27 expression and the phosphorylation of Smad2, p44/p42 MAPK, p38 MAPK, and SAPK/JNK were evaluated by western blot analysis.

Result

HSP90 inhibitors 17-dimethylaminoethylamino-17-demethoxy-geldanamycin (17-DMAG) and onalespib significantly enhanced the TGF-β-induced HSP27 expression. TGF-β inhibitor SB431542 reduced the enhancement by 17-DMAG or onalespib of the TGF-β-induced HSP27 expression levels. HSP90 inhibitors, geldanamycin, onalespib, and 17-DMAG did not affect the TGF-β-stimulated phosphorylation of Smad2. Geldanamycin did not affect the TGF-β-stimulated phosphorylation of p44/p42 MAPK or p38 MAPK but significantly enhanced the TGF-β-stimulated phosphorylation of SAPK/JNK. Onalespib also increased the TGF-β-stimulated phosphorylation of SAPK/JNK. Furthermore, SP600125, a specific inhibitor for SAPK/JNK, significantly suppressed onalespib or geldanamycin’s enhancing effect of the TGF-β-induced HSP27 expression levels.

Conclusion

Our results strongly suggest that HSP90 inhibitors upregulated the TGF-β-induced HSP27 expression and that these effects of HSP90 inhibitors were mediated through SAPK/JNK pathway in osteoblasts.
Appendix
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Literature
1.
Kampinga HH, Hageman J, Vos MJ, Kubota H, Tanguay RM, Bruford EA, et al. Guidelines for the nomenclature of the human heat shock proteins. Cell Stress Chaperones. 2009;14:105–11.CrossRef
2.
Hang K, Ye C, Chen E, Zhang W, Xue D, Pan Z. Role of the heat shock protein family in bone metabolism. Cell Stress Chaperones. 2018;23:1153–64.CrossRef
3.
Sreedhar AS, Kalmar E, Csermely P, Shen YF. Hsp90 isoforms: functions, expression and clinical importance. FEBS Lett. 2004;562:11–5.CrossRef
4.
Whitesell L, Lindquist SL. HSP90 and the chaperoning of cancer. Nat Rev Cancer. 2005;5:761–72.CrossRef
5.
Trepel J, Mollapour M, Giaccone G, Neckers L. Targeting the dynamic HSP90 complex in cancer. Nat Rev Cancer. 2010;10:537–49.CrossRef
6.
Rong B, Yang S. Molecular mechanism and targeted therapy of Hsp90 involved in lung cancer: new discoveries and developments (review). Int J Oncol. 2018;52:321–36.PubMed
7.
Haque A, Alam Q, Alam MZ, Azhar EI, Sait KH, Anfinan N, et al. Current understanding of HSP90 as a novel therapeutic target: an emerging approach for the treatment of Cancer. Curr Pharm Des. 2016;22:2947–59.CrossRef
8.
Xu W, Neckers L. Targeting the molecular chaperone heat shock protein 90 provides a multifaceted effect on diverse cell signaling pathways of cancer cells. Clin Cancer Res. 2007;13:1625–9.CrossRef
9.
Fuhrmann-Stroissnigg H, Ling YY, Zhao J, McGowan SJ, Zhu Y, Brooks RW, et al. Identification of HSP90 inhibitors as a novel class of senolytics. Nat Commun. 2017;8:422.CrossRef
10.
Sims NA, Morris HA, Moore RJ, Durbridge TC. Increased bone resorption precedes increased bone formation in the ovariectomized rat. Calcif Tissue Int. 1996;59:121–7.CrossRef
11.
Martin TJ, Sims NA. Osteoclast-derived activity in the coupling of bone formation to resorption. Trends Mol Med. 2005;11:76–81.CrossRef
12.
Morikawa M, Derynck R, Miyazono K. TGF-β and the TGF-β family: context-dependent roles in cell and tissue physiology. Cold Spring Harb Perspect Biol. 2016;8:pii: a021873.CrossRef
13.
Wu M, Chen G, Li YP. TGF-β and BMP signaling in osteoblast, skeletal development, and bone formation, homeostasis and disease. Bone Res. 2016;4:16009.CrossRef
14.
Zhang YE. Non-Smad pathways in TGF-β signaling. Cell Res. 2009;19:128–39.CrossRef
15.
Hatakeyama D, Kozawa O, Niwa M, Matsuno H, Ito H, Kato K, et al. Upregulation by retinoic acid of transforming growth factor-β-stimulated heat shock protein 27 induction in osteoblasts: involvement of mitogen-activated protein kinases. Biochim Biophys Acta. 2002;1589:15–30.CrossRef
16.
Hayashi K, Takai S, Matsushima-Nishiwaki R, Hanai Y, Kato K, Tokuda H, et al. (−)-Epigallocatechin gallate reduces transforming growth factor β-stimulated HSP27 induction through the suppression of stress-activated protein kinase/c-Jun N-terminal kinase in osteoblasts. Life Sci. 2008;82:1012–7.CrossRef
17.
Kainuma S, Tokuda H, Yamamoto N, Kuroyanagi G, Fujita K, Kawabata T, et al. Heat shock protein 27 (HSPB1) suppresses the PDGF-BB-induced migration of osteoblasts. Int J Mol Med. 2017;40:1057–66.CrossRef
18.
Kato K, Adachi S, Matsushima-Nishiwaki R, Minamitani C, Natsume H, Katagiri Y, et al. Regulation by heat shock protein 27 of osteocalcin synthesis in osteoblasts. Endocrinology. 2011;152:1872–82.CrossRef
19.
Romanello M, Bivi N, Pines A, Deganuto M, Quadrifoglio F, Moro L, et al. Bisphosphonates activate nucleotide receptors signaling and induce the expression of Hsp90 in osteoblast-like cell lines. Bone. 2006;39:739–53.CrossRef
20.
Miyasaka M, Nakata H, Hao J, Kim YK, Kasugai S, Kuroda S. Low-intensity pulsed ultrasound stimulation enhances heat-shock protein 90 and mineralized nodule formation in mouse calvaria-derived osteoblasts. Tissue Eng Part A. 2015;21:2829–39.CrossRef
21.
Fujita K, Otsuka T, Kawabata T, Sakai G, Matsushima-Nishiwaki R, Kozawa O, et al. Inhibitors of heat shock protein 90 augment endothelin-1-induced heat shock protein 27 through the SAPK/JNK signaling pathway in osteoblasts. Mol Med Rep. 2018;17:8542–7.PubMed
22.
Kim W, Tokuda H, Kawabata T, Fujita K, Sakai G, Nakashima D, et al. Enhancement by HSP90 inhibitor of PGD2-stimulated HSP27 induction in osteoblasts: suppression of SAPK/JNK and p38 MAP kinase. Prostaglandins Other Lipid Mediat. 2019;143:106327.CrossRef
23.
Fujita K, Otsuka T, Kawabata T, Kainuma S, Sakai G, Matsushima-Nishiwaki R, et al. HSP90 limits thrombin-stimulated IL-6 synthesis in osteoblast-like MC3T3-E1 cells: regulation of p38 MAPK. Int J Mol Med. 2018;42:2185–92.PubMed
24.
Sudo H, Kodama H, Amagai Y, Yamamoto S, Kasai S. In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol. 1982;96:191–8.CrossRef
25.
Kozawa O, Tokuda H, Miwa M, Kotoyori J, Oiso Y. Cross-talk regulation between cyclic-AMP production and phosphoinositide hydrolysis induced by prostaglandin E2 in osteoblast-like cells. Exp Cell Res. 1992;198:130–4.CrossRef
26.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680–5.CrossRef
27.
Egorin MJ, Lagattuta TF, Hamburger DR, Covey JM, White KD, Musser SM, et al. Pharmacokinetics, tissue distribution, and metabolism of 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (NSC 707545) in CD2F1 mice and Fischer 344 rats. Cancer Chemother Pharmacol. 2002;49:7–19.CrossRef
28.
Woodhead AJ, Angove H, Carr MG, Chessari G, Congreve M, Coyle JE, et al. Discovery of (2,4-dihydroxy-5-isopropylphenyl)-[5-(4-methylpiperazin-1-ylmethyl)-1,3-dihydroisoindol-2-yl] methanone (AT13387), a novel inhibitor of the molecular chaperone Hsp90 by fragment based drug design. J Med Chem. 2010;53:5956–69.CrossRef
29.
Yamamoto N, Otsuka T, Kondo A, Matsushima-Nishiwaki R, Kuroyanagi G, Kozawa O, et al. Rac limits TGF-β-induced VEGF synthesis in osteoblasts. Mol Cell Endocrinol. 2015;405:35–41.CrossRef
30.
Ochel HJ, Eichhorn K, Gademann G. Geldanamycin: the prototype of a class of antitumor drugs targeting the heat shock protein 90 family of molecular chaperones. Cell Stress Chaperones. 2001;6:105–12.CrossRef
31.
Bennett BL, Sasaki DT, Murray BW, O'Leary EC, Sakata ST, Xu W, et al. SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc Natl Acad Sci U S A. 2001;98:13681–6.CrossRef
32.
Samadi AK, Zhang X, Mukerji R, Donnelly AC, Blagg BS, Cohen MS. A novel C-terminal HSP90 inhibitor KU135 induces apoptosis and cell cycle arrest in melanoma cells. Cancer Lett. 2011;312:158–67.CrossRef
33.
Smith V, Sausville EA, Camalier FR, Fiebig HH, Burger AM. Comparison of 17-dimethylaminoethylamino-17-demethoxy-geldanamycin (17DMAG) and 17-allylamino-17-demethoxygeldanamycin (17AAG) in vitro: effects on Hsp90 and client proteins in melanoma models. Cancer Chemother Pharmacol. 2005;56:126–37.CrossRef
34.
Mellatyar H, Talaei S, Pilehvar-Soltanahmadi Y, Barzegar A, Akbarzadeh A, Shahabi A, et al. Targeted cancer therapy through 17-DMAG as an Hsp90 inhibitor: overview and current state of the art. Biomed Pharmacother. 2018;102:608–17.CrossRef
35.
Chen H, Xing J, Hu X, Chen L, Lv H, Xu C, et al. Inhibition of heat shock protein 90 rescues glucocorticoid-induced bone loss through enhancing bone formation. J Steroid Biochem Mol Biol. 2017;171:236–46.CrossRef
Metadata
Title
Upregulation of TGF-β-induced HSP27 by HSP90 inhibitors in osteoblasts
Authors
Gen Kuroyanagi
Haruhiko Tokuda
Kazuhiko Fujita
Tetsu Kawabata
Go Sakai
Woo Kim
Tomoyuki Hioki
Junko Tachi
Rie Matsushima-Nishiwaki
Takanobu Otsuka
Hiroki Iida
Osamu Kozawa
Publication date
01-12-2022
Publisher
BioMed Central
Keywords
Shock
Shock
Published in
BMC Musculoskeletal Disorders / Issue 1/2022
Electronic ISSN: 1471-2474
DOI
https://doi.org/10.1186/s12891-022-05419-1

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