Anti-tumor activity of a miR-199-dependent oncolytic adenovirus.
The down-regulation of miR-199 occurs in nearly all primary hepatocellular carcinomas (HCCs) and HCC cell lines in comparison with normal liver. We exploited this miR-199 differential expression to develop a conditionally replication-competent oncolytic adenovirus, Ad-199T, and achieve tumor-specific viral expression and replication. To this aim, we introduced four copies of miR-199 target sites within the 3' UTR of E1A gene, essential for viral replication. As consequence, E1A expression from Ad-199T virus was tightly regulated both at RNA and protein levels in HCC derived cell lines, and replication controlled by the level of miR-199 expression. Various approaches were used to asses in vivo properties of Ad-199T. Ad-199T replication was inhibited in normal, miR-199 positive, liver parenchyma, thus resulting in reduced hepatotoxicity. Conversely, the intrahepatic delivery of Ad-199T in newborn mice led to virus replication and fast removal of implanted HepG2 liver cancer cells. The ability of Ad-199T to control tumor growth was also shown in a subcutaneous xenograft model in nude mice and in HCCs arising in immune-competent mice. In summary, we developed a novel oncolytic adenovirus, Ad-199T, which could demonstrate a therapeutic potential against liver cancer without causing significant hepatotoxicity.
Dipartimento di Morfologia, Chirurgia e Medicina Sperimentale, Università di Ferrara, Ferrara, Italy.
Elisa
E
Callegari
Bahaeldin K
BK
Elamin
Lucilla
L
D'Abundo
Simonetta
S
Falzoni
Giovanna
G
Donvito
Farzaneh
F
Moshiri
Maddalena
M
Milazzo
Giuseppe
G
Altavilla
Luciano
L
Giacomelli
Francesca
F
Fornari
Akseli
A
Hemminki
Francesco
F
Di Virgilio
Laura
L
Gramantieri
Massimo
M
Negrini
Silvia
S
Sabbioni
D020413:3' Untranslated Regions; D000256:Adenoviridae / Q000235:genetics*; D017500:Adenovirus E1A Proteins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000401:mortality / Q000473:pathology / Q000628:therapy; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D015967:Gene Expression Regulation, Viral; D023061:Gene Order; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000401:mortality / Q000473:pathology / Q000628:therapy; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D050130:Oncolytic Virotherapy; D050504:Oncolytic Viruses / Q000235:genetics*; D034622:RNA Interference; D047368:Tumor Burden / Q000235:genetics; D014779:Virus Replication; D023041:Xenograft Model Antitumor Assays
D020413:3' Untranslated Regions; D017500:Adenovirus E1A Proteins; D035683:MicroRNAs; C531408:mirn199 microRNA, human