In vitro and in vivo characterization of a novel Hedgehog signaling antagonist in human glioblastoma cell lines

Int J Cancer. 2012 Jul 15;131(2):E33-44. doi: 10.1002/ijc.27349. Epub 2012 Jan 31.

Abstract

Glioblastoma multiforme (GBM) is composed of heterogeneous and genetically different cells, which are highly invasive and motile. The standard chemotherapeutic agent, temozolomide, affects GBM cell proliferation but is generally unable to prevent tumor recurrence. Hedgehog pathway activation has been reported to be relevant in GBM and different pharmacological pathway modulators have been identified. We report that by growing a commercially available recurrent GBM cell line (DBTRG-05MG) without serum and in the presence of defined growth factors; we obtained a less differentiated cell population, growing in suspension as neurospheres, in which the Hedgehog pathway is activated. Furthermore, the expression profile of Hedgehog pathway components found in DBTRG-05MG neurospheres is similar to primary stem-like cells derived from recurrent GBM patients. We report the effect of our novel specific Smoothened receptor antagonist (SEN450) on neurosphere growing cells and compared its effect to that of well known benchmark compounds. Finally, we showed that SEN450 is both antiproliferative on its own and further reduces tumor volume after temozolomide pretreatment in a mouse xenograft model using DBTRG-05MG neurosphere cells. Altogether our data indicate that the Hedgehog pathway is not irreversibly switched off in adherent cells but can be reactivated when exposed to well-defined culture conditions, thus restoring the condition observed in primary tumor-derived material, and that pharmacological modulation of this pathway can have profound influences on tumor proliferation. Therefore, pharmacological inhibition of the Hedgehog pathway is a potentially useful therapeutic approach in GBM.

MeSH terms

  • Anilides / pharmacology
  • Animals
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Dacarbazine / analogs & derivatives
  • Dacarbazine / pharmacology
  • Gene Expression Profiling
  • Glioblastoma / drug therapy*
  • Glioblastoma / genetics
  • Glioblastoma / metabolism*
  • Glioblastoma / pathology
  • Hedgehog Proteins / antagonists & inhibitors*
  • Hedgehog Proteins / metabolism
  • Humans
  • Mice
  • Mice, Nude
  • Pyridines / pharmacology
  • Receptors, G-Protein-Coupled / antagonists & inhibitors*
  • Signal Transduction / drug effects*
  • Smoothened Receptor
  • Temozolomide
  • Transcription Factors / metabolism
  • Veratrum Alkaloids / pharmacology
  • Xenograft Model Antitumor Assays
  • Zinc Finger Protein GLI1

Substances

  • Anilides
  • GLI1 protein, human
  • Hedgehog Proteins
  • HhAntag691
  • Pyridines
  • Receptors, G-Protein-Coupled
  • SMO protein, human
  • Smoothened Receptor
  • Transcription Factors
  • Veratrum Alkaloids
  • Zinc Finger Protein GLI1
  • Dacarbazine
  • Temozolomide
  • cyclopamine