title: Impairment of glioma stem cell survival and growth by a novel inhibitor for Survivin-Ran protein complex.
abstract: OBJECTIVE
  Glioblastoma multiforme (GBM) is a devastating disease. Recent studies suggest that the stem cell properties of GBM contribute to the development of therapy resistance.
  
  
  METHODS
  The expression of Survivin and Ran was evaluated by immunohistochemistry with GBM tissues, and quantitative reverse transcriptase (qRT)-PCR and immunocytochemistry with patient-derived GBM sphere cultures. With a computational structure-based drug design, 11 small-molecule compounds were designed, synthesized, and evaluated as inhibitor candidates for the molecular interaction of Survivin protein. The molecular mechanism of the lead compound, LLP-3, was determined by Western blot, ELISA, in situ proximity ligation assay, and immunocytochemistry. The effects of LLP-3 treatment on GSCs were evaluated both in vitro and in vivo. Quantitative immunohistochemistry was carried out to compare Survivin expression in tissues from 44 newly diagnosed and 31 recurrent post-chemoradiation GBM patients. Lastly, the sensitivities of temozolomide-resistant GBM spheres to LLP-3 were evaluated in vitro.
  
  
  RESULTS
  Survivin and Ran were strongly expressed in GBM tissues, particularly in the perivasculature, and also in patient-derived GSC cultures. LLP-3 treatment disrupted the Survivin-Ran protein complex in cancer cells and abolished the growth of patient-derived GBM spheres in vitro and in vivo. This inhibition was dependent on caspase activity and associated with p53 status of cells. Immunohistochemistry showed that Survivin expression is significantly increased in recurrent GBM compared with newly diagnosed tumors, and temozolomide-resistant GBM spheres exhibited high sensitivities to LLP-3 treatment.
  
  
  CONCLUSIONS
  Disruption of the Survivin-Ran complex by LLP-3 abolishes survival and growth of GSCs both in vitro and in vivo, indicating an attractive novel therapeutic approach for GBM.
authors.affiliation: Department of Neurological Surgery, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Hacer
authors.initials: H
authors.lastname: Guvenc
authors.forename: Marat S
authors.initials: MS
authors.lastname: Pavlyukov
authors.forename: Kaushal
authors.initials: K
authors.lastname: Joshi
authors.forename: Habibe
authors.initials: H
authors.lastname: Kurt
authors.forename: Yeshavanth K
authors.initials: YK
authors.lastname: Banasavadi-Siddegowda
authors.forename: Ping
authors.initials: P
authors.lastname: Mao
authors.forename: Christopher
authors.initials: C
authors.lastname: Hong
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Yamada
authors.forename: Chang-Hyuk
authors.initials: CH
authors.lastname: Kwon
authors.forename: Deepak
authors.initials: D
authors.lastname: Bhasin
authors.forename: Somsundaram
authors.initials: S
authors.lastname: Chettiar
authors.forename: Gaspar
authors.initials: G
authors.lastname: Kitange
authors.forename: In-Hee
authors.initials: IH
authors.lastname: Park
authors.forename: Jann N
authors.initials: JN
authors.lastname: Sarkaria
authors.forename: Chenglong
authors.initials: C
authors.lastname: Li
authors.forename: Mihail I
authors.initials: MI
authors.lastname: Shakhparonov
authors.forename: Ichiro
authors.initials: I
authors.lastname: Nakano
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D019008:Drug Resistance, Neoplasm; D005909:Glioblastoma / Q000378:metabolism* / Q000401:mortality / Q000601:surgery; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D011379:Prognosis; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation; D055503:Protein Multimerization; D015398:Signal Transduction; D000077022:Survivin; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays; D020931:ran GTP-Binding Protein / Q000737:chemistry / Q000378:metabolism*
chemical_list: C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D000077022:Survivin; D016159:Tumor Suppressor Protein p53; D020931:ran GTP-Binding Protein