Recombinant adenovirus with increased safety and anticancer activities, and use thereof
The present invention relates to a recombinant adenovirus with increased in-vivo safety, tissue specificity, and anticancer activities, and a use thereof. Specifically, the recombinant adenovirus comprising: a promoter of the liver tissue-specific phosphoenolpyruvate carboxykinase (PEPCK) gene; a trans-splicing ribozyme which is operably linked to the promoter and acts on a cancer-specific gene; a therapeutic gene or a reporter gene which is linked to the 3′ exon of the ribozyme; and a serotype 35 fiber knob and a serotype 5 shaft, in which the orf4 gene is deleted from adenovirus E1, E3 and E4 orf1, shows remarkable in-vivo safety, high specificity for a target tissue, and remarkable anticancer effects, and thus can be useful for an anticancer drug or a cancer diagnostic agent as a gene delivery vector.
1. A recombinant adenovirus comprising: a nucleic acid sequence construct comprising a liver specific phosphoenolpyruvate carboxykinase (PEPCK) gene promoter, a trans-splicing ribozyme gene operably linked to the promoter, a therapeutic gene or a reporter gene linked to the 3′ exon of the trans-splicing ribozyme gene, a gene enhancer, and a serotype 35 fiber knob and serotype 5 shaft gene, the recombinant adenovirus lacking adenovirus E1, E3 and E4 orf1 to orf4 genes,
wherein the gene enhancer is an ApoE (apolipoprotein E) gene enhancer having the nucleotide sequence of SEQ ID NO: 6, a PEPCK gene enhancer, a serum albumin gene enhancer, an AFP (alphafetoprotein) gene enhancer, a CEA (carcinoembryonic antigen) gene enhancer, or a PSA (prostate-specific antigen) gene enhancer; and
wherein the trans-splicing ribozyme gene has the nucleotide sequence of SEQ ID NO: 3.
2. The recombinant adenovirus of claim 1 , wherein the liver specific PEPCK gene promoter has the nucleotide sequence of SEQ ID NO: 1.
3. The recombinant adenovirus of claim 1 , wherein the promoter further comprises an ApoA1 intron having the nucleotide sequence of SEQ ID NO: 2.
4. The recombinant adenovirus of claim 1 , wherein the therapeutic gene is selected from the group consisting of drug-sensitizing genes, proapoptotic genes, cytostatic genes, cytotoxic genes, tumor suppressor genes, antigenic genes, cytokine genes, and anti-angiogenic genes.
5. The recombinant adenovirus of claim 1 , wherein the reporter gene is a gene that encodes LacZ, CAT (chloramphenicol acetyl transferase), Renila luciferase, firefly luciferase, red fluorescent protein (RFP), green fluorescent protein (GFP), secreted placental alkaline phosphatase (SEAP) or HSV-tk (Herpes simplex virus-thymidine kinase).
6. The recombinant adenovirus of claim 1 , wherein the therapeutic gene is a HSV-tk (herpes simplex virus-thymidine kinase) gene having the nucleotide sequence of SEQ ID NO: 4.
7. A pharmaceutical composition for treating liver cancer, which comprises, as an active ingredient, the recombinant adenovirus set forth in claim 1 .
8. A composition for diagnosing liver cancer, which comprises, as an active ingredient, the recombinant adenovirus set forth in claim 1 .