Synergetic cytotoxic activity toward breast cancer cells enhanced by the combination of Antp-TPR hybrid peptide targeting Hsp90 and Hsp70-targeted peptide

BMC Cancer

Tomohisa Horibe¹  Aya Torisawa¹  Masayuki Kohno¹  Koji Kawakami¹ 

[1] Department of Pharmacoepidemiology, Graduate School of Medicine and Public Health, Kyoto University, Yoshida Konoecho, 606-8501 Sakyo-ku, Kyoto, Japan

关键词: Bioluminescence;    Breast cancer;    Molecular chaperone;    Hybrid peptide;    Heat shock protein 70;    Heat shock protein 90;    Client proteins;   

Others  :  1134696
DOI  :  10.1186/1471-2407-14-615

 received in 2013-12-02, accepted in 2014-08-21,  发布年份 2014

【 摘 要 】

Background

Heat shock protein (Hsp) 90 and Hsp70 are indispensable for cell survival under conditions of stress. They bind to client proteins to assist in protein stabilization, translocation of polypeptides across the cell membrane, and recovery of proteins from aggregates in the cell. Therefore, these proteins have recently emerged as important targets in the treatment of cancer. We previously reported that the newly designed Antp-TPR hybrid peptide targeting Hsp90 induced cytotoxic activity to cancer cells both in vitro and in vivo.

Methods

To further improve the cytotoxic activity of Antp-TPR toward cancer cells, we investigated the effect of a Hsp70-targeted peptide, which was made cell-permeable by adding the polyarginine with a linker sequence, on the cytotoxic activity of Antp-TPR in breast cancer cell lines.

Results

It was revealed that Antp-TPR in the presence of a Hsp70-targeted peptide induced effective cytotoxic activity toward breast cancer cells through the descrease of Hsp90 client proteins such as p53, Akt, and cRaf. Moreover, the combined treatment with these peptides did not induce the up-regulation of Hsp70 protein, as determined by western blotting, a promoter assay using a luminometer, and single-cell level imaging with the LV200 system, although a small-molecule inhibitor of Hsp90, 17-allylamino-demethoxygeldanamycin (17-AAG), did induce the up-regulation of this protein. We also found that treatment with Antp-TPR, Hsp70-targeted peptide, or a combination of the two did not induce an increase in the glutathione concentrations in the cancer cells.

Conclusion

These findings suggest that targeting both Hsp90 and Hsp70 with Antp-TPR and Hsp70-targeted peptide is an attractive approach for selective cancer cell killing that might provide potent and selective therapeutic options for the treatment of cancer.

【 授权许可】

   
2014 Horibe et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin 2011, 61:69-90.
• [2]Workman P, Burriws F, Neckers L, Rosen N: Drugging the cancer chaperone HSP90: combinatorial therapeutic exploitation of oncogene addiction and tumor stress. Ann N Y Acad Sci 2007, 1113:202-216.
• [3]Chiosis G, Timaul MN, Lucas B, Munster PN, Zheng FF, Sepp-Lorenzino L, Rosen N: A small molecule designed to bind to the adenine nucleotide pocket of Hsp90 causes Her2 degradation and the growth arrest and differentiation of breast cancer cells. Chem Biol 2001, 8:289-299.
• [4]Modi S, Stopeck AT, Gordon MS, Mendelson D, Solit DB, Bagatell R, Ma W, Wheler J, Rosen N, Norton L, Cropp GF, Johnson RG, Hannah AL, Hudis CA: Combination of trastuzumab and tanespimycin (17-AAG, KOS-953) is safe and active in trastuzumab-refractory HER-2 overexpressing breast cancer: a phase I dose-escalation study. J Clin Oncol 2007, 25:5410-5417.
• [5]Caldas-Lopes E, Cerchietti L, Ahn JH, Clement CC, Robles AI, Rodina A, Moulick K, Taldone T, Gozman A, Guo Y, Wu N, de Stanchina E, White J, Gross SS, Ma Y, Varticovski L, Melnick A, Chiosis G: Hsp90 inhibitor PU-H71, a multimodal inhibitor of malignancy, induces complete responses in triple-negative breast cancer models. Proc Natl Acad Sci U S A 2009, 106:8368-8373.
• [6]Röhl A, Rohrberg J, Buchner J: The chaperone Hsp90: changing partners for demanding clients. Trends Biochem Sci 2013, 38:253-262.
• [7]Soga S, Akinaga S, Shiotsu Y: Hsp90 inhibitors as anti-cancer agents, from basic discoveries to clinical development. Curr Pharm Des 2013, 19:366-376.
• [8]Maloney A, Workman P: HSP90 as a new therapeutic target for cancer therapy: the story unfolds. Expert Opin Biol Ther 2002, 2:3-24.
• [9]Garcia-Carbonero R, Carnero A, Paz-Ares L: Inhibition of HSP90 molecular chaperones: moving into the clinic. Lancet Oncol 2013, 14:e358-369.
• [10]Li J, Buchner J: Structure, function and regulation of the hsp90 machinery. Biomed J 2013, 36:106-117.
• [11]Powers MV, Jones K, Barillari C, Westwood I, van Montfort RL, Workman P: Targeting HSP70: the second potentially druggable heat shock protein and molecular chaperone? Cell Cycle 2010, 9:1542-1550.
• [12]Rérole AL, Gobbo J, De Thonel A, Schmitt E, de Pais Barros JP, Hammann A, Lanneau D, Fourmaux E, Deminov O, Micheau O, Lagrost L, Colas P, Kroemer G, Garrido C: Peptides and aptamers targeting HSP70: a novel approach for anticancer chemotherapy. Cancer Res 2011, 71:484-495.
• [13]Horibe T, Kohno M, Haramoto M, Ohara K, Kawakami K: Designed hybrid TPR peptide targeting Hsp90 as a novel anticancer agent. J Transl Med 2011, 9:8. BioMed Central Full Text
• [14]Horibe T, Torisawa A, Kohno M, Kawakami K: Molecular mechanism of cytotoxicity induced by Hsp90-targeted Antp-TPR hybrid peptide in glioblastoma cells. Mol Cancer 2012, 11:59. BioMed Central Full Text
• [15]Leu JI, Pimkina J, Frank A, Murphy ME, George DL: A small molecule inhibitor of inducible heat shock protein 70. Mol Cell 2009, 36:15-27.
• [16]Leu JI, Pimkina J, Pandey P, Murphy ME, George DL: Heat shock protein-70 inhibition by the small-molecule 2-phenylethynesulfonamide impairs protein clearance pathways in tumor cells. Mol Cancer Res 2011, 9:936-947.
• [17]Kabouridis PS: Biological application of protein transduction technology. Trends Biotechnol 2003, 21:498-503.
• [18]Futaki S, Suzuki T, Ohashi W, Yagami T, Tanaka S, Ueda K, Sugiura Y: Arginine-rich peptides. J Biol Chem 2001, 276:5836-5840.
• [19]Futaki S: Membrane-permeable arginine-rich peptides and the translocation mechanism. Adv Drug Deliv Rev 2005, 57:547-558.
• [20]Kohno M, Horibe T, Haramoto M, Yano Y, Ohara K, Nakajima O, Matsuzaki K, Kawakami K: A novel hybrid peptide targeting EGFR-expressing cancers. Eur J Cancer 2011, 47:773-783.
• [21]Saito S, Furuno A, Sakurai J, Sakamoto A, Park HR, Shin-ya K, Tsuruno T, Tomida A: Chemical genomics identifies the unfolded protein response as a target for selective cancer cell killing during glucose deprivation. Cancer Res 2009, 69:4225-4234.
• [22]Akiyoshi R, Suzuki H: The measurement of cytosolic ATP during apoptosis: Bioluminescence imaging at the single cell level. In Bioluminescence and Chemiluminescence. Volume 15. Edited by Shen X, Yang XL, Zhang XR, Cui ZJ, Kricka LJ, Stanley PE. World Scientific; 2008::339-342.
• [23]Horibe T, Torisawa A, Akiyoshi R, Hatta-Ohashi Y, Suzuki H, Kawakami K: Transfection efficiency of normal and cancer cell lines and monitoring of promoter activity by single-cell bioluminescence imaging. Luminescence 2014, 29:96-100.
• [24]Zhang H, Chung D, Yang YC, Neely L, Tsurumoto S, Fan J, Zhang L, Biamonte M, Brekken J, Lundgren K, Burrows F: Identification of new biomarkers for clinical trials of Hsp90 inhibitors. Mol Cancer Ther 2006, 5:1256-1264.
• [25]Song D, Chaerkady R, Tan AC, García-García E, Nalli A, Suárez-Gauthier A, López-Ríos F, Zhang XF, Solomon A, Tong J, Read M, Fritz C, Jimeno A, Pandey A, Hidalgo M: Antitumor activity and molecular effects of the novel heat shock protein 90 inhibitor, IPI-504, in pancreatic cancer. Mol Cancer Ther 2008, 7:3275-3284.
• [26]McCollum AK, TenEyck CJ, Sauer BM, Toft DO, Erlichman C: Up-regulation of heat shock protein 27 induces resistance to 17-allylamino-demethoxygeldanamycin through a glutathione-mediated mechanism. Cancer Res 2006, 66:10967-10975.
• [27]Kawamoto M, Horibe T, Kohno M, Kawakami K: Her2 targeted hybrid peptide that blocks her2 tyrosine kinase disintegrates cancer cell membrane and inhibits tumor growth in vivo. Mol Cancer Ther 2013, 12:384-393.
• [28]Hollestelle A, Elstrodt F, Nagel JH, Kallemeijn WW, Schutte M: Phosphatidylinositol-3-OH kinase or RAS pathway mutations in human breast cancer cell lines. Mol Cancer Res 2007, 5:195-201.
• [29]O’Brien NA, Browne BC, Chow L, Wang Y, Ginther C, Arboleda J, Duffy MJ, Crown J, O’Donovan N, Slamon DJ: Activated phosphoinositide 3-kinase/AKT signaling confers resistance to trastuzumab but not lapatinib. Mol Cancer Ther 2010, 9:1489-1502.
• [30]Jhaveri K, Modi S: Hsp90 inhibitors for cancer therapy and overcoming drug resistance. Adv Pharmacol 2012, 65:471-517.
• [31]Zamaraeva MV, Sabirov RZ, Maeno E, Ando-Akatsuka Y, Bessonova SV, Okada Y: Cells die with increased cytosolic ATP during apoptosis: a bioluminescence study with intracellular luciferase. Cell Death Differ 2005, 12:1390-1397.
• [32]Schlecht R, Scholz SR, Dahmen H, Wegener A, Sirrenberg C, Musil D, Bomke J, Eggenweiler HM, Mayer MP, Bukau B: Functional analysis of Hsp70 inhibitors. PLoS One 2013, 8:e78443.
• [33]Granato M, Lacconi V, Peddis M, Lotti LV, Renzo LD, Gonnella R, Trivedi P, Frati L, D’Orazi G, Faggioni A, Cirone M: HSP70 inhibition by 2-phenylethynesulfonamide induces lysosomal cathepsin D release and immunogenic cell death in primary effusion lymphoma. Cell Death Dis 2013, 4:e730.