Radiation-induced upregulation of gene expression from adenoviral vectors mediated by DNA damage repair and regulation.
OBJECTIVE
In the present study, we evaluated the combination of replication-deficient adenoviruses and radiotherapy in vitro. The purpose of the present study was to analyze the mechanism of radiation-mediated upregulation of adenoviral transgene expression.


METHODS
Adenoviral transgene expression (luciferase or green fluorescent protein) was studied with and without radiation in three cell lines: breast cancer M4A4-LM3, prostate cancer PC-3MM2, and lung cancer LNM35/enhanced green fluorescent protein. The effect of the radiation dose, modification of the viral capsid, and five different transgene promoters were studied. The cellular responses were studied using mass spectrometry and immunofluorescence analysis. Double strand break repair was modulated by inhibitors of heat shock protein 90, topoisomerase-I, and DNA protein kinase, and transgene expression was measured.


RESULTS
We found that a wide range of radiation doses increased adenoviral transgene expression regardless of the cell line, transgene, promoter, or viral capsid modification. Treatment with adenovirus, radiation, and double strand break repair inhibitors resulted in persistence of double strand breaks and subsequent increases in adenovirus transgene expression.


CONCLUSIONS
Radiation-induced enhancement of adenoviral transgene expression is linked to DNA damage recognition and repair. Radiation induces a global cellular response that results in increased production of RNA and proteins, including adenoviral transgene products. This study provides a mechanistic rationale for combining radiation with adenoviral gene delivery.
Cancer Gene Therapy Group, Molecular Cancer Biology Program, Transplantation Laboratory, Haartman Institute, University of Helsinki, Helsinki, Finland.
Petri
P
Nokisalmi
Maria
M
Rajecki
Sari
S
Pesonen
Sophie
S
Escutenaire
Rabah
R
Soliymani
Mikko
M
Tenhunen
Laura
L
Ahtiainen
Akseli
A
Hemminki
D000260:Adenoviruses, Human / Q000235:genetics / Q000528:radiation effects*; D045744:Cell Line, Tumor; D003131:Combined Modality Therapy / Q000379:methods; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D003673:Defective Viruses / Q000235:genetics / Q000528:radiation effects*; D015870:Gene Expression / Q000235:genetics / Q000528:radiation effects*; D015967:Gene Expression Regulation, Viral / Q000235:genetics / Q000528:radiation effects; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000528:radiation effects*; D049452:Green Fluorescent Proteins / Q000235:genetics; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D008156:Luciferases / Q000235:genetics; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011829:Radiation Dosage; D059004:Topoisomerase I Inhibitors / Q000494:pharmacology; D019076:Transgenes / Q000235:genetics / Q000528:radiation effects*; D015854:Up-Regulation / Q000235:genetics / Q000528:radiation effects; D014779:Virus Replication / Q000235:genetics / Q000528:radiation effects
D018841:HSP90 Heat-Shock Proteins; D047428:Protein Kinase Inhibitors; D059004:Topoisomerase I Inhibitors; D049452:Green Fluorescent Proteins; D008156:Luciferases