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. 2012:797:111-25.
doi: 10.1007/978-1-61779-340-0_9.

Targeting brain tumor stem cells with oncolytic adenoviruses

Marta M Alonso  1 , Hong Jiang, Candelaria Gomez-Manzano, Juan Fueyo
Affiliations

Targeting brain tumor stem cells with oncolytic adenoviruses

Marta M Alonso et al. Methods Mol Biol. 2012.

Abstract

In 2004, brain tumor stem cells (BTSCs) were isolated from surgical human malignant gliomas. This cancer cell population has been identified as the root for tumor initiation and resistance to therapies. Thus, it is imperative to develop new therapies that can eradicate this subpopulation to improve the prognosis of patients with brain tumors. Our group previously reported the antiglioma effect of the tumor-selective oncolytic adenovirus Delta-24-RGD that is now being tested in a phase I clinical trial for patients with malignant gliomas. We also showed that Delta-24-RGD infects, replicates in, and induces cell death in BTSCs. Interestingly, we observed that adenoviral-infected cells undergo autophagy and that autophagy-related cytoplasmic vacuolization might be part of the lysis process. Here, we summarize the materials and methods used in our study as follows: establishment of neurosphere cultures from surgical samples of human glioblastoma multiformes; assessment of stem cell markers; examination of adenoviral receptors in BTSCs; evaluation of the cytotoxicity induced by oncolytic adenoviruses; and assessment of autophagy in oncolytic adenovirus-infected BTSCs in vitro, and finally we describe a method to detect upregulation of the autophagy-related protein Atg5 in tumors treated with Delta-24-RGD.

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Figures

Fig. 1
Fig. 1
Neurosphere cultures were established from acute cell dissociation of human glioblastoma multiforme surgical specimens and maintained in neuro-stem cell medium. Note that the cells grow as spheroids. This figure has been modified from a previous publication (9).
Fig. 2
Fig. 2
Electron microscopy photographs showing (a) mock-infected and (b) Delta-24-RGD-infected BTSCs. Please note that cytoplasmic vacuoles are present in the virus-infected cells but not in the mock-infected cells. Close-up images of the Delta-24-RGD-infected cells in (b) show (c) a cluster of Delta-24-RGD virions (white arrow) in the nucleus and (d ) complex autophagic vacuolar bodies in the cytoplasm. This figure has been modified from a previous publication (9).
See this image and copyright information in PMC

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