title: Antitumor effects of minodronate, a third-generation nitrogen-containing bisphosphonate, in synergy with γδT cells in human glioblastoma in vitro and in vivo.
abstract: Nitrogen-containing bisphosphonates (N-BPs), which prevent bone resorption, exert direct and γδT cell (GDT)-mediated antitumor effects against several tumor cell types, including glioblastoma (GBM). However, limited information is available regarding the antitumor effects of N-BPs in GBM. Specifically, the antitumor effects of minodronate (MDA), a third-generation N-BP, in GBM are yet unclear. This study aimed to investigate the antitumor effects of MDA in GBM in vitro and in vivo. We performed growth inhibition and apoptosis detection assays using the GBM cell lines U87MG and U138MG. Apoptosis inhibition assays were also conducted. In vivo xenograft assays were performed in highly immunodeficient NOD.Cg-Prkdc(scid) Il2rg(tm1Sug)/Jic mice subcutaneously implanted with U87MG and U138MG cells. Growth inhibition and apoptosis detection assays demonstrated that MDA inhibited GBM cell growth via apoptosis, which was markedly enhanced by ex vivo expanded GDT. A pan-caspase inhibitor, z-VAD-fmk, inhibited MDA-induced U138MG apoptosis and MDA/GDT-induced U87MG and U138MG apoptosis. But z-VAD-fmk increased MDA-induced U87MG apoptosis. MDA/GDT-mediated apoptosis was blocked by the anti-T cell receptor (TCR) Vγ9, mevalonate pathway inhibitor, granzyme B inhibitor, and antitumor necrosis factor (TNF)-α. In vivo xenograft assays showed that combined intraperitoneal administration of MDA/GDT induced antitumor effects on unestablished U87MG-derived subcutaneous tumors. MDA exerted direct and GDT-mediated anti-GBM apoptotic effects in a caspase-dependent manner. GDT recognized MDA-exposed GBM cells via TCRVγ9 and induced apoptosis via granzyme B and TNF-α release. Because MDA elicited anti-GBM effects in synergy with GDT in vivo, a combination of MDA and ex vivo-generated GDT could be an effective treatment in patients with GBM.
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan. nakazawa@naramed-u.ac.jp.|Grandsoul Research Institute for Immunology, Inc., 8-1, Matsui, Uda, Nara, 633-2221, Japan. nakazawa@naramed-u.ac.jp.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nakazawa
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Nakamura
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Matsuda
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Nishimura
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Young Soo
authors.initials: YS
authors.lastname: Park
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Motoyama
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Hironaka
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Ichiro
authors.initials: I
authors.lastname: Nakagawa
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yokota
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Yamada
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Tamura
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takeshima
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Kouji
authors.initials: K
authors.lastname: Omoto
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Tanaka
authors.affiliation: Department of Pathogen, Infection and Immunity, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Yukiteru
authors.initials: Y
authors.lastname: Ouji
authors.affiliation: Department of Pathogen, Infection and Immunity, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Masahide
authors.initials: M
authors.lastname: Yoshikawa
authors.affiliation: Clinic Grandsoul Nara, 8-1, Matsui, Uda, Nara, 633-2221, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tsujimura
authors.affiliation: Department of Neurosurgery, Nara Medical University School of Medicine, 480, Shijocho, Kashihara, Nara, 634-8522, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nakase
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D001932:Brain Neoplasms / Q000628:therapy*; D061945:Caspase Inhibitors / Q000494:pharmacology; D002452:Cell Count; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004164:Diphosphonates / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D005909:Glioblastoma / Q000628:therapy*; D006801:Humans; D007093:Imidazoles / Q000627:therapeutic use*; D000075942:Intraepithelial Lymphocytes / Q000502:physiology* / Q000637:transplantation*; D008297:Male; D016688:Mice, Inbred NOD; D015398:Signal Transduction / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D017304:Annexin A5; D000970:Antineoplastic Agents; D061945:Caspase Inhibitors; D004164:Diphosphonates; D007093:Imidazoles; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C087958:YM 529
keywords: Apoptosis; Glioblastoma; Minodronate; NOG mouse; γδT cells