The HDAC Inhibitors Scriptaid and LBH589 Combined with the Oncolytic Virus Delta24-RGD Exert Enhanced Anti-Tumor Efficacy in Patient-Derived Glioblastoma Cells.

Abstract

BACKGROUND:

A phase I/II trial for glioblastoma with the oncolytic adenovirus Delta24-RGD was recently completed. Delta24-RGD conditionally replicates in cells with a disrupted retinoblastoma-pathway and enters cells via αvβ3/5 integrins. Glioblastomas are differentially sensitive to Delta24-RGD. HDAC inhibitors (HDACi) affect integrins and share common cell death pathways with Delta24-RGD. We studied the combination treatment effects of HDACi and Delta24-RGD in patient-derived glioblastoma stem-like cells (GSC), and we determined the most effective HDACi.

METHODS:

SAHA, Valproic Acid, Scriptaid, MS275 and LBH589 were combined with Delta24-RGD in fourteen distinct GSCs. Synergy was determined by Chou Talalay method. Viral infection and replication were assessed using luciferase and GFP encoding vectors and hexon-titration assays. Coxsackie adenovirus receptor and αvβ3 integrin levels were determined by flow cytometry. Oncolysis and mechanisms of cell death were studied by viability, caspase-3/7, LDH and LC3B/p62, phospho-p70S6K. Toxicity was studied on normal human astrocytes. MGMT promotor methylation status, TCGA classification, Rb-pathway and integrin gene expression levels were assessed as markers of responsiveness.

RESULTS:

Scriptaid and LBH589 acted synergistically with Delta24-RGD in approximately 50% of the GSCs. Both drugs moderately increased αvβ3 integrin levels and viral infection in responding but not in non-responding GSCs. LBH589 moderately increased late viral gene expression, however, virus titration revealed diminished viral progeny production by both HDACi, Scriptaid augmented caspase-3/7 activity, LC3B conversion, p62 and phospho-p70S6K consumption, as well as LDH levels. LBH589 increased LDH and phospho-p70S6K consumption. Responsiveness correlated with expression of various Rb-pathway genes and integrins. Combination treatments induced limited toxicity to human astrocytes.

CONCLUSION:

LBH589 and Scriptaid combined with Delta24-RGD revealed synergistic anti-tumor activity in a subset of GSCs. Both HDACi moderately augmented viral infection and late gene expression, but slightly reduced progeny production. The drugs differentially activated multiple cell death pathways. The limited toxicity on astrocytes supports further evaluation of the proposed combination therapies.

• Authors: Lotte M E Berghauser Pont, Anne Kleijn, Jenneke J Kloezeman, Wouter van den Bossche, Johanna K Kaufmann, Jeroen de Vrij, Sieger Leenstra, Clemens M F Dirven, Martine L M Lamfers
• Journal: PloS one (PLoS One) Vol. 10 Issue 5 Pg. e0127058 ( 2015) ISSN: 1932-6203 [Electronic] United States
• PMID: 25993039 (Publication Type: Journal Article)

Chemical References

• Histone Deacetylase Inhibitors
• Hydroxamic Acids
• Hydroxylamines
• Indoles
• Integrin alphaVbeta3
• Quinolines
• Tumor Suppressor Protein p53
• Tumor Suppressor Proteins
• scriptaid
• Panobinostat
• DNA Modification Methylases
• MGMT protein, human
• DNA Repair Enzymes

Topics

• Adenoviridae (genetics)
• Animals
• Apoptosis (drug effects)
• Astrocytes (drug effects, metabolism)
• Autophagy (drug effects)
• Brain Neoplasms (genetics, pathology, therapy)
• Cell Survival
• DNA Methylation
• DNA Modification Methylases (genetics)
• DNA Repair Enzymes (genetics)
• Disease Models, Animal
• Dose-Response Relationship, Drug
• Glioblastoma (genetics, pathology, therapy)
• Histone Deacetylase Inhibitors (pharmacology)
• Humans
• Hydroxamic Acids (pharmacology)
• Hydroxylamines (pharmacology)
• Indoles (pharmacology)
• Integrin alphaVbeta3 (metabolism)
• Mice
• Mutation
• Neoplastic Stem Cells (metabolism, pathology)
• Oncolytic Virotherapy
• Oncolytic Viruses
• Panobinostat
• Promoter Regions, Genetic
• Quinolines (pharmacology)
• Tumor Suppressor Protein p53 (metabolism)
• Tumor Suppressor Proteins (genetics)
• Virus Replication (drug effects)
• Xenograft Model Antitumor Assays