[en] Pro-survival stress-inducible chaperone HSP110 is the only HSP for which a mutation has been found in a cancer. Multicenter clinical studies demonstrated a direct association between HSP110 inactivating mutation presence and excellent prognosis in colorectal cancer patients. Here, we have combined crystallographic studies on human HSP110 and in silico modeling to identify HSP110 inhibitors that could be used in colorectal cancer therapy. Two molecules (foldamers 33 and 52), binding to the same cleft of HSP110 nucleotide-binding domain, were selected from a chemical library (by co-immunoprecipitation, AlphaScreening, Interference-Biolayer, Duo-link). These molecules block HSP110 chaperone anti-aggregation activity and HSP110 association to its client protein STAT3, thereby inhibiting STAT3 phosphorylation and colorectal cancer cell growth. These effects were strongly decreased in HSP110 knockdown cells. Foldamer's 33 ability to inhibit tumor growth was confirmed in two colorectal cancer animal models. Although tumor cell death (apoptosis) was noted after treatment of the animals with foldamer 33, no apparent toxicity was observed, notably in epithelial cells from intestinal crypts. Taken together, we identified the first HSP110 inhibitor, a possible drug-candidate for colorectal cancer patients whose unfavorable outcome is associated to HSP110.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Gozzi, Gustavo J; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Gonzalez, Daniel; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Boudesco, Christophe; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Dias, Alexandre M M; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Gotthard, Guillaume; European Synchrotron Radiation Facility, F-38043, Grenoble, France.
Uyanik, Burhan; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Dondaine, Lucile; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Marcion, Guillaume ; Université de Liège - ULiège > GIGA > GIGA I3 - Hematology ; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Hermetet, François; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Denis, Camille; Normandie Université, UNICAEN, EA 4258 CERMN (Centre d'Etudes et de Recherche sur
Hardy, Laurianne; Normandie Université, UNICAEN, EA 4258 CERMN (Centre d'Etudes et de Recherche sur
Suzanne, Peggy; Normandie Université, UNICAEN, EA 4258 CERMN (Centre d'Etudes et de Recherche sur
Douhard, Romain; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Jego, Gaetan; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Dubrez, Laurence ; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Demidov, Oleg N; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France.
Neiers, Fabrice; University of Burgundy Franche-Comté, Dijon, France. ; Centre des Sciences du Goût et de l'Alimentation, INRA, CNRS, Dijon, France.
Briand, Loïc; University of Burgundy Franche-Comté, Dijon, France. ; Centre des Sciences du Goût et de l'Alimentation, INRA, CNRS, Dijon, France.
Sopková-de Oliveira Santos, Jana; Normandie Université, UNICAEN, EA 4258 CERMN (Centre d'Etudes et de Recherche sur
Voisin-Chiret, Anne-Sophie; Normandie Université, UNICAEN, EA 4258 CERMN (Centre d'Etudes et de Recherche sur
Garrido, Carmen; INSERM UMR1231, Laboratory of Excellence LipSTIC and label Ligue Nationale contre ; University of Burgundy Franche-Comté, Dijon, France. cgarrido@u-bourgogne.fr. ; Georges François Leclerc Center (CGFL), Dijon, France. cgarrido@u-bourgogne.fr.
Factsheet of WHO 2012 on cancer available here: http://www.cancerresearchuk.org/sites/default/files/cs_report_world.pdf
Le DT, Uram JN, Wang H, Bartlett BR, Kemberling H, Eyring AD, et al. PD-1 blockade in tumors with mismatch-repair deficiency. New Engl J Med. 2015;372:2509–20.
Chung KY, Gore I, Fong L, Venook A, Beck SB, Dorazio P, et al. Phase II study of the anti-cytotoxic T-lymphocyte-associated antigen 4 monoclonal antibody, tremelimumab, in patients with refractory metastatic colorectal cancer. J Clin Oncol. 2010;28:3485–90.
Champiat S, Dercle L, Ammari S, Massard C, Hollebecque A, Postel-Vinay S, et al. Hyperprogressive disease is a new pattern of progression in cancer patients treated by anti-PD-1/PD-L1. Clin Cancer Res. 2017;23:1920–8.
Berthenet K, Boudesco C, Collura A, Svrcek M, Richaud S, Hammann A, et al. Extracellular HSP110 skews macrophage polarization in colorectal cancer. Oncoimmunology. 2016;5:e1170264.
Berthenet K, Bokhari A, Lagrange A, Marcion G, Boudesco C, Causse S, et al. HSP110 promotes colorectal cancer growth through STAT3 activation. Oncogene. 2017;36:2328–36.
Guttmann DM, Koumenis C. The heat shock proteins as targets for radiosensitization and chemosensitization in cancer. Cancer Biol Ther. 2011;12:1023–31.
Calderwood SK, Khaleque MA, Sawyer DB, Ciocca DR. Heat shock proteins in cancer: chaperones of tumorigenesis. Trends Biochem Sci. 2006;31:164–72.
Schmitt E, Maingret L, Puig PE, Rerole AL, Ghiringhelli F, Hammann A, et al. Heat shock protein 70 neutralization exerts potent antitumor effects in animal models of colon cancer and melanoma. Cancer Res. 2006;66:4191–7.
Bonniaud P, Burgy O, Garrido C. Heat shock protein-90 toward theranostics: a breath of fresh air in idiopathic pulmonary fibrosis. Eur Respir J. 2018;51:1702612.
Collura A, Lagrange A, Svrcek M, Marisa L, Buhard O, Guilloux A, et al. Patients with colorectal tumors with microsatellite instability and large deletions in HSP110 T17 have improved response to 5-fluorouracil-based chemotherapy. Gastroenterology. 2014;146:401–11 e401.
Rampelt H, Kirstein-Miles J, Nillegoda NB, Chi K, Scholz SR, Morimoto RI, et al. Metazoan Hsp70 machines use Hsp110 to power protein disaggregation. EMBO J. 2012;31:4221–35.
Mattoo RU, Sharma SK, Priya S, Finka A, Goloubinoff P. Hsp110 is a bona fide chaperone using ATP to unfold stable misfolded polypeptides and reciprocally collaborate with Hsp70 to solubilize protein aggregates. J Biol Chem. 2013;288:21399–411.
Slaby O, Sobkova K, Svoboda M, Garajova I, Fabian P, Hrstka R, et al. Significant overexpression of Hsp110 gene during colorectal cancer progression. Oncol Rep. 2009;21:1235–41.
Zappasodi R, Ruggiero G, Guarnotta C, Tortoreto M, Tringali C, Cavane A, et al. HSPH1 inhibition downregulates Bcl-6 and c-Myc and hampers the growth of human aggressive B-cell non-Hodgkin lymphoma. Blood. 2015;125:1768–71.
Hwang TS, Han HS, Choi HK, Lee YJ, Kim YJ, Han MY, et al. Differential, stage-dependent expression of Hsp70, Hsp110 and Bcl-2 in colorectal cancer. J Gastroenterol Hepatol. 2003;18:690–700.
Dorard C, de Thonel A, Collura A, Marisa L, Svrcek M, Lagrange A, et al. Expression of a mutant HSP110 sensitizes colorectal cancer cells to chemotherapy and improves disease prognosis. Nat Med. 2011;17:1283–9.
Yu N, Kakunda M, Pham V, Lill JR, Du P, Wongchenko M, et al. HSP105 recruits protein phosphatase 2A to dephosphorylate beta-catenin. Mol Cell Biol. 2015;35:1390–1400.
Flaherty KM, DeLuca-Flaherty C, McKay DB. Three-dimensional structure of the ATPase fragment of a 70K heat-shock cognate protein. Nature. 1990;346:623–8.
Liu Q, Hendrickson WA. Insights into Hsp70 chaperone activity from a crystal structure of the yeast Hsp110 Sse1. Cell. 2007;131:106–20.
Santos JS, Voisin-Chiret AS, Burzicki G, Sebaoun L, Sebban M, Lohier JF, et al. Structural characterizations of oligopyridyl foldamers, alpha-helix mimetics. J Chem Inf Model. 2012;52:429–39.
Raviol H, Sadlish H, Rodriguez F, Mayer MP, Bukau B. Chaperone network in the yeast cytosol: Hsp110 is revealed as an Hsp70 nucleotide exchange factor. EMBO J. 2006;25:2510–8.
Joly AL, Deepti A, Seignez A, Goloudina A, Hebrard S, Schmitt E, et al. The HSP90 inhibitor, 17AAG, protects the intestinal stem cell niche and inhibits graft versus host disease development. Oncogene. 2016;35:2948.
Dragovic Z, Broadley SA, Shomura Y, Bracher A, Hartl FU. Molecular chaperones of the Hsp110 family act as nucleotide exchange factors of Hsp70s. EMBO J. 2006;25:2519–28.
Raviol H, Bukau B, Mayer MP. Human and yeast Hsp110 chaperones exhibit functional differences. FEBS Lett. 2006;580:168–74.
Oh HJ, Easton D, Murawski M, Kaneko Y, Subjeck JR. The chaperoning activity ofhsp110. Identification of functional domains by use of targeted deletions. J Biol Chem. 1999;274:15712–8.
Yamagishi N, Ishihara K, Hatayama T. Hsp105alpha suppresses Hsc70 chaperone activity by inhibiting Hsc70 ATPase activity. J Biol Chem. 2004;279:41727–33.
Andreasson C, Fiaux J, Rampelt H, Mayer MP, Bukau B. Hsp110 is a nucleotide-activated exchange factor for Hsp70. J Biol Chem. 2008;283:8877–84.
Buhard O, Lagrange A, Guilloux A, Colas C, Chouchene M, Wanherdrick K, et al. HSP110 T17 simplifies and improves the microsatellite instability testing in patients with colorectal cancer. J Med Genet. 2016;53:377–84.
Kimura A, Ogata K, Altan B, Yokobori T, Ide M, Mochiki E, et al. Nuclear heat shock protein 110 expression is associated with poor prognosis and chemotherapy resistance in gastric cancer. Oncotarget. 2016;7:18415–23.
Kimura A, Ogata K, Altan B, Yokobori T, Mochiki E, Yanai M, et al. Nuclear heat shock protein 110 expression is associated with poor prognosis and hyperthermo-chemotherapy resistance in gastric cancer patients with peritoneal metastasis. World J Gastroenterol. 2017;23:7541–50.
Scott DE, Bayly AR, Abell C, Skidmore J. Small molecules, big targets: drug discovery faces the protein-protein interaction challenge. Nat Rev Drug Discov. 2016;15:533–50.
Morikawa T, Baba Y, Yamauchi M, Kuchiba A, Nosho K, Shima K, et al. STAT3 expression, molecular features, inflammation patterns, and prognosis in a database of 724 colorectal cancers. Clin Cancer Res. 2011;17:1452–62.
Huang W, Dong Z, Chen Y, Wang F, Wang CJ, Peng H, et al. Small-molecule inhibitors targeting the DNA-binding domain of STAT3 suppress tumor growth, metastasis and STAT3 target gene expression in vivo. Oncogene. 2016;35:802.
Welte T, Zhang SS, Wang T, Zhang Z, Hesslein DG, Yin Z, et al. STAT3 deletion during hematopoiesis causes Crohn’s disease-like pathogenesis and lethality: a critical role of STAT3 in innate immunity. Proc Natl Acad Sci USA. 2003;100:1879–84.
Mantel C, Messina-Graham S, Moh A, Cooper S, Hangoc G, Fu XY, et al. Mouse hematopoietic cell-targeted STAT3 deletion: stem/progenitor cell defects, mitochondrial dysfunction, ROS overproduction, and a rapid aging-like phenotype. Blood. 2012;120:2589–99.
Shen Y, Guo D, Weng L, Wang S, Ma Z, Yang Y, et al. Tumor-derived exosomes educate dendritic cells to promote tumor metastasis via HSP72/HSP105-TLR2/TLR4 pathway. Oncoimmunology. 2017;6:e1362527.
Veber DF, Johnson SR, Cheng HY, Smith BR, Ward KW, Kopple KD. Molecular properties that influence the oral bioavailability of drug candidates. J Med Chem. 2002;45:2615–23.
Martin YC. A bioavailability score. J Med Chem. 2005;48:3164–70.
Daina A, Zoete V. A boiled-egg to predict gastrointestinal absorption and brain penetration of small molecules. ChemMedChem. 2016;11:1117–21.
Nurizzo D, Mairs T, Guijarro M, Rey V, Meyer J, Fajardo P, et al. The ID23-1 structural biology beamline at the ESRF. J Synchrotron Radiat. 2006;13(Pt 3):227–38.
Emsley P, Cowtan K. Coot: model-building tools for molecular graphics. Acta Crystallogr Sect D, Biol Crystallogr. 2004;60(Pt 12 Pt 1):2126–32.
Adams PD, Afonine PV, Bunkoczi G, Chen VB, Davis IW, Echols N, et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr Sect D, Biol Crystallogr. 2010;66(Pt 2):213–21.
Perrakis A, Harkiolaki M, Wilson KS, Lamzin VS. ARP/wARP and molecular replacement. Acta Crystallogr Sect D, Biol Crystallogr. 2001;57(Pt 10):1445–50.
Murshudov GN, Skubak P, Lebedev AA, Pannu NS, Steiner RA, Nicholls RA, et al. REFMAC5 for the refinement of macromolecular crystal structures. Acta Crystallogr Sect D, Biol Crystallogr. 2011;67(Pt 4):355–67.
Chen VB, Arendall WB 3rd, Headd JJ, Keedy DA, Immormino RM, Kapral GJ, et al. MolProbity: all-atom structure validation for macromolecular crystallography. Acta Crystallogr Sect D, Biol Crystallogr. 2010;66(Pt 1):12–21.
DeLano WL. The PyMOL Molecular Graphics System, Version 2.0. Schrödinger, LLC. http://www.pymol.org.
Ashkenazy H, Abadi S, Martz E, Chay O, Mayrose I, Pupko T, et al. ConSurf 2016: an improved methodology to estimate and visualize evolutionary conservation in macromolecules. Nucleic Acids Res. 2016;44(W1):W344–350.
