title: Anti-Tumor Effects of Second Generation β-Hydroxylase Inhibitors on Cholangiocarcinoma Development and Progression.
abstract: Cholangiocarcinoma (CCA) has a poor prognosis due to widespread intrahepatic spread. Aspartate β-hydroxylase (ASPH) is a transmembrane protein and catalyzes the hydroxylation of aspartyl and asparaginyl residues in calcium binding epidermal growth factor (cbEGF)-like domains of various proteins, including Notch receptors and ligands. ASPH is highly overexpressed (>95%) in human CCA tumors. We explored the molecular mechanisms by which ASPH mediated the CCA malignant phenotype and evaluated the potential of ASPH as a therapeutic target for CCA. The importance of expression and enzymatic activity of ASPH for CCA growth and progression was examined using shRNA "knockdown" and a mutant construct that reduced its catalytic activity. Second generation small molecule inhibitors (SMIs) of β-hydroxylase activity were developed and used to target ASPH in vitro and in vivo. Subcutaneous and intrahepatic xenograft rodent models were employed to determine anti-tumor effects on CCA growth and development. It was found that the enzymatic activity of ASPH was critical for mediating CCA progression, as well as inhibiting apoptosis. Mechanistically, ASPH overexpression promoted Notch activation and modulated CCA progression through a Notch1-dependent cyclin D1 pathway. Targeting ASPH with shRNAs or a SMI significantly suppressed CCA growth in vivo.
authors.affiliation: Liver Research Center, Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Chiung-Kuei
authors.initials: CK
authors.lastname: Huang
authors.affiliation: Liver Research Center, Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Iwagami
authors.affiliation: Liver Research Center, Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Arihiro
authors.initials: A
authors.lastname: Aihara
authors.affiliation: Liver Research Center, Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Waihong
authors.initials: W
authors.lastname: Chung
authors.affiliation: Liver Research Center, Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Suzanne
authors.initials: S
authors.lastname: de la Monte
authors.affiliation: Department of Medical Chemistry, College of Pharmacy Glendale, Midwestern University, Glendale, Arizona, United States of America.
authors.forename: John-Michael
authors.initials: JM
authors.lastname: Thomas
authors.affiliation: Department of Medical Chemistry, College of Pharmacy Glendale, Midwestern University, Glendale, Arizona, United States of America.
authors.forename: Mark
authors.initials: M
authors.lastname: Olsen
authors.affiliation: Liver Research Center, Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Rolf
authors.initials: R
authors.lastname: Carlson
authors.affiliation: Liver Research Center, Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Tunan
authors.initials: T
authors.lastname: Yu
authors.affiliation: Department of Biomedical and Pharmaceutical Science, College of Pharmacy, The University of Rhode Island, Pharmacy Building, 7 Greenhouse Road, Kingston, Rhode Island, United States of America.
authors.forename: Xiaoqun
authors.initials: X
authors.lastname: Dong
authors.affiliation: Liver Research Center, Warren Alpert Medical School of Brown University and Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Jack
authors.initials: J
authors.lastname: Wands
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D002135:Calcium-Binding Proteins / Q000037:antagonists & inhibitors*; D045744:Cell Line, Tumor; D018281:Cholangiocarcinoma / Q000188:drug therapy / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000037:antagonists & inhibitors*; D051379:Mice; D008819:Mice, Nude; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors*; D009124:Muscle Proteins / Q000037:antagonists & inhibitors*; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D002135:Calcium-Binding Proteins; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D009124:Muscle Proteins; D006899:Mixed Function Oxygenases; C419389:ASPH protein, human