title: Dual Targeting of Epithelial Ovarian Cancer Via Folate Receptor α and the Proton-Coupled Folate Transporter with 6-Substituted Pyrrolo[2,3-d]pyrimidine Antifolates.
abstract: Folate uptake in epithelial ovarian cancer (EOC) involves the reduced folate carrier (RFC) and the proton-coupled folate transporter (PCFT), both facilitative transporters and folate receptor (FR) α. Although in primary EOC specimens, FRα is widely expressed and increases with tumor stage, PCFT was expressed independent of tumor stage (by real-time RT-PCR and IHC). EOC cell line models, including cisplatin sensitive (IGROV1 and A2780) and resistant (SKOV3 and TOV112D) cells, expressed a 17-fold range of FRα and similar amounts (within ∼2-fold) of PCFT. Novel 6-substituted pyrrolo[2,3-d]pyrimidine thienoyl antifolatesAGF94andAGF154exhibited potent antiproliferative activities toward all of the EOC cell lines, reflecting selective cellular uptake by FRα and/or PCFT over RFC. When IGROV1 cells were pretreated withAGF94at pH 6.8, clonogenicity was potently inhibited, confirming cell killing. FRα was knocked down in IGROV1 cells with lentiviral shRNAs. Two FRα knockdown clones (KD-4 and KD-10) showed markedly reduced binding and uptake of [3H]folic acid and [3H]AGF154by FRα, but maintained high levels of [3H]AGF154uptake by PCFT compared to nontargeted control cells. In proliferation assays, KD-4 and KD-10 cells preservedin vitroinhibition byAGF94andAGF154, compared to a nontargeted control, attributable to residual FRα- and substantial PCFT-mediated uptake. KD-10 tumor xenografts in severe-compromised immune-deficient mice were likewise sensitive toAGF94Collectively, our results demonstrate the substantial therapeutic potential of novel 6-substituted pyrrolo[2,3-d]pyrimidine antifolates with dual targeting of PCFT and FRα toward EOCs that express a range of FRα, along with PCFT, as well as cisplatin resistance.Mol Cancer Ther; 16(5); 819-30. ©2017 AACR.
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan. houz@karmanos.org matherly@karmanos.org gangjee@duq.edu.|Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Zhanjun
authors.initials: Z
authors.lastname: Hou
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Leda
authors.initials: L
authors.lastname: Gattoc
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Carrie
authors.initials: C
authors.lastname: O'Connor
authors.affiliation: Division of Medicinal Chemistry, Graduate School of Pharmaceutical Science, Duquesne University, Pittsburgh, Pennsylvania.
authors.forename: Si
authors.initials: S
authors.lastname: Yang
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Adrianne
authors.initials: A
authors.lastname: Wallace-Povirk
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Christina
authors.initials: C
authors.lastname: George
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Steve
authors.initials: S
authors.lastname: Orr
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.|Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Lisa
authors.initials: L
authors.lastname: Polin
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Kathryn
authors.initials: K
authors.lastname: White
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Juiwanna
authors.initials: J
authors.lastname: Kushner
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan.|Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Robert T
authors.initials: RT
authors.lastname: Morris
authors.affiliation: Division of Medicinal Chemistry, Graduate School of Pharmaceutical Science, Duquesne University, Pittsburgh, Pennsylvania. houz@karmanos.org matherly@karmanos.org gangjee@duq.edu.
authors.forename: Aleem
authors.initials: A
authors.lastname: Gangjee
authors.affiliation: Department of Oncology, Wayne State University School of Medicine, Detroit, Michigan. houz@karmanos.org matherly@karmanos.org gangjee@duq.edu.|Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute, Detroit, Michigan.|Department of Pharmacology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Larry H
authors.initials: LH
authors.lastname: Matherly
mesh_terms: D000818:Animals; D000077216:Carcinoma, Ovarian Epithelial; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin / Q000008:administration & dosage; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D058975:Folate Receptor 1 / Q000037:antagonists & inhibitors / Q000235:genetics*; D005492:Folic Acid / Q000235:genetics; D005493:Folic Acid Antagonists / Q000008:administration & dosage* / Q000138:chemical synthesis; D006801:Humans; D051379:Mice; D009375:Neoplasms, Glandular and Epithelial / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D058979:Proton-Coupled Folate Transporter / Q000235:genetics*; D023041:Xenograft Model Antitumor Assays
chemical_list: D058975:Folate Receptor 1; D005493:Folic Acid Antagonists; D058979:Proton-Coupled Folate Transporter; C505058:SLC46A1 protein, human; D005492:Folic Acid; D002945:Cisplatin