title: UDP glucuronosyltransferase 1A expression levels determine the response of colorectal cancer cells to the heat shock protein 90 inhibitor ganetespib.
abstract: HSP90 inhibition represents a promising route to cancer therapy, taking advantage of cancer cell-inherent proteotoxic stress. The HSP90-inhibitor ganetespib showed benefit in advanced clinical trials. This raises the need to identify the molecular determinants of treatment response. We tested the efficacy of ganetespib on a series of colorectal cancer (CRC)-derived cell lines and correlated their sensitivities with comprehensive gene expression analysis. Notably, the drug concentration required for 50% growth inhibition (IC50) varied up to 70-fold (from 36 to 2500 nM) between different cell lines. Correlating cell line-specific IC50s with the corresponding gene expression patterns revealed a strong association between ganetespib resistance (IC50>500 nM) and high expression of the UDP glucuronosyltransferase 1A (UGT1A) gene cluster. Moreover, CRC tumor samples showed a comparable distribution of UGT1A expression levels. The members of the UGT1A gene family are known as drug-conjugating liver enzymes involved in drug excretion, but their function in tumor cells is hardly understood. Chemically unrelated HSP90 inhibitors, for example, 17-N-allylamino-17-demethoxygeldanamycin (17-AAG), did not show correlation of drug sensitivities with UGT1A levels, whereas the ganetespib-related compound NVP-AUY922 did. When the most ganetespib-resistant cell line, HT29, was treated with ganetespib, the levels of HSP90 clients were unaffected. However, HT29 cells became sensitized to the drug, and HSP90 client proteins were destabilized by ganetespib upon siRNA-mediated UGT1A knockdown. Conversely, the most ganetespib-sensitive cell lines HCT116 and SW480 became more tolerant toward ganetespib upon UGT1A overexpression. Mechanistically, ganetespib was rapidly glucuronidated and excreted in resistant but not in sensitive CRC lines. We conclude that CRC cell-expressed UGT1A inactivates ganetespib and other resorcinolic Hsp90 inhibitors by glucuronidation, which renders the drugs unable to inhibit Hsp90 and thereby abrogates their biological activity. UGT1A levels in tumor tissues may be a suitable predictive biomarker to stratify CRC patients for ganetespib treatment.
authors.affiliation: Göttingen Centre of Molecular Biosciences (GZMB), Institute of Molecular Oncology, Faculty of Medicine, University of Göttingen, 37077 Göttingen, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Landmann
authors.affiliation: Synta Pharmaceuticals Corp, 45 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: D A
authors.initials: DA
authors.lastname: Proia
authors.affiliation: Synta Pharmaceuticals Corp, 45 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: S
authors.initials: S
authors.lastname: He
authors.affiliation: Synta Pharmaceuticals Corp, 45 Hartwell Avenue, Lexington, MA 02421, USA.
authors.forename: L S
authors.initials: LS
authors.lastname: Ogawa
authors.affiliation: Department of Medical Statistics, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Kramer
authors.affiliation: Department of Medical Statistics, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Beißbarth
authors.affiliation: Department of General, Visceral and Pediatric Surgery, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Grade
authors.affiliation: Department of General, Visceral and Pediatric Surgery, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Gaedcke
authors.affiliation: Department of General, Visceral and Pediatric Surgery, University Medical Center Göttingen, Göttingen, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Ghadimi
authors.affiliation: 1] Göttingen Centre of Molecular Biosciences (GZMB), Institute of Molecular Oncology, Faculty of Medicine, University of Göttingen, 37077 Göttingen, Germany [2] Department of Pathology, School of Medicine, Stony Brook University, Stony Brook, NY 11794, USA.
authors.forename: U
authors.initials: U
authors.lastname: Moll
authors.affiliation: Göttingen Centre of Molecular Biosciences (GZMB), Institute of Molecular Oncology, Faculty of Medicine, University of Göttingen, 37077 Göttingen, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Dobbelstein
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000188:drug therapy / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D014230:Triazoles / Q000494:pharmacology*; D000099148:UGT1A1 Enzyme
chemical_list: D000970:Antineoplastic Agents; D014453:Glucuronosyltransferase; D018841:HSP90 Heat-Shock Proteins; D014230:Triazoles; D000099148:UGT1A1 Enzyme; C533237:STA 9090