Ionizing radiation-induced adenovirus infection is mediated by Dynamin 2.
Specific viral targeting into intrahepatic tumors remains critical for adenovirus gene therapy in liver cancer. We previously showed that ionizing radiation increases adenovirus uptake and transgene expression in cells and colon cancer xenografts. Here, we tested whether radiation induces viral uptake through virus-cell membrane interaction. We found that radiation (8 Gy) induced adenoviral gene transfer in rat hepatocytes (WB) and human colon carcinoma cells (LoVo). This induction (24.4- and 6.5-fold, respectively) and viral uptake were significantly diminished by preincubation with antibody for Dynamin 2 but not for Coxsackie adenovirus receptor or for integrin alpha(v). Radiation-induced Dynamin 2 expression was detected by immunohistochemical staining and by increased mRNA levels for Dynamin 2 in WB (1.5-fold) and LoVo (2.2-fold) cells. Specific small interference RNA (siRNA) transfection significantly inhibited Dynamin 2 expression in various tumor cell lines (LoVo, D54, and MCF-7) and abolished the radiation induction of Dynamin 2. Likewise, radiation-induced viral gene transfer in these cells (6.5-, 5.5-, and 9.0-fold, respectively) was significantly reduced in siRNA-transfected cells (2.7-, 3.7-, and 5.0-fold, respectively). Moreover, viral uptake in LoVo tumor xenografts was significantly increased in s.c. tumors (10.9-fold) when adenovirus was given i.v. at 24 hours after tumor irradiation, coincident with an elevated Dynamin 2 expression in irradiated tumors. These data suggest that ionizing radiation induces adenovirus gene transfer in cells and tumor xenografts by regulating viral uptake, potentially through interaction with cellular Dynamin 2 and thus should provide insight into improving adenovirus targeting in tumors.
Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Jun
J
Qian
Jiong
J
Yang
Aleksandar F
AF
Dragovic
Eyad
E
Abu-Isa
Theodore S
TS
Lawrence
Ming
M
Zhang
D000256:Adenoviridae / Q000235:genetics / Q000472:pathogenicity / Q000528:radiation effects*; D000818:Animals; D001932:Brain Neoplasms / Q000235:genetics / Q000628:therapy / Q000821:virology; D001943:Breast Neoplasms / Q000235:genetics / Q000628:therapy / Q000821:virology; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics / Q000628:therapy / Q000821:virology; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D034285:Dynamin II / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D039564:Integrin alphaV / Q000502:physiology; D051379:Mice; D008819:Mice, Nude; D009369:Neoplasms / Q000235:genetics / Q000628:therapy / Q000821:virology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D011991:Receptors, Virus / Q000502:physiology; D014162:Transfection; D023041:Xenograft Model Antitumor Assays
C568771:CLMP protein, human; C568772:CLMP protein, mouse; C568773:Clmp protein, rat; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D039564:Integrin alphaV; D034741:RNA, Small Interfering; D011991:Receptors, Virus; D034285:Dynamin II