Nanoparticle mediated silencing of DNA repair sensitizes pediatric brain tumor cells to y-irradiation

Authors

Forrest M. Kievit, University of WashingtonFollow
Zachary R. Stephen, University of Washington, Seattle
Kui Wang, University of Washington, Seattle
Christopher J. Dayringer, University of Washington, Seattle
Jonathan G. Sham, University of Washington, Seattle
Richard G. Ellenbogen, University of Washington, SeattleFollow
John R. Silber, University of Washington, Seattle
Miqin Zhang, University of Washington, SeattleFollow

Date of this Version

2015

Citation

Molecular Oncology 9 (2015) 1071-1080

Comments

Copyright 2015 Federation of European Biochemical Societies.

http://dx.doi.org/10.1016/j.molonc.2015.01.006

Abstract

Medulloblastoma (MB) and ependymoma (EP) are the most common pediatric brain tumors, afflicting 3000 children annually. Radiotherapy (RT) is an integral component in the treatment of these tumors; however, the improvement in survival is often accompanied by radiation-induced adverse developmental and psychosocial sequelae. Therefore, there is an urgent need to develop strategies that can increase the sensitivity of brain tumors cells to RT while sparing adjacent healthy brain tissue. Apurinic endonuclease 1 (Ape1), an enzyme in the base excision repair pathway, has been implicated in radiation resistance in cancer. Pharmacological and specificity limitations inherent to small molecule inhibitors of Ape1 have hindered their clinical development. Here we report on a nanoparticle (NP) based siRNA delivery vehicle for knocking down Ape1 expression and sensitizing pediatric brain tumor cells to RT. The NP comprises a superparamagnetic iron oxide core coated with a biocompatible, biodegradable coating of chitosan, polyethylene glycol (PEG), and polyethyleneimine (PEI) that is able to bind and protect siRNA from degradation and to deliver siRNA to the perinuclear region of target cells. NPs loaded with siRNA against Ape1 (NP:siApe1) knocked down Ape1 expression over 75% in MB and EP cells, and reduced Ape1 activity by 80%. This reduction in Ape1 activity correlated with increased DNA damage post-irradiation, which resulted in decreased cell survival in clonogenic assays. The sensitization was specific to therapies generating abasic lesions as evidenced by NP:siRNA not increasing sensitivity to paclitaxel, a microtubule disrupting agent. Our results indicate NP-mediated delivery of siApe1 is a promising strategy for circumventing pediatric brain tumor resistance to RT.