title: Development of Recombinant Adeno-Associated Virus Serotype 2/8 Carrying Kringle Domains of Human Plasminogen for Sustained Expression and Cancer Therapy.
abstract: Angiostatin and other plasminogen derivatives exhibit antitumor activities directly or indirectly, have demonstrated promising anticancer effects in preclinical studies, but have mostly failed in clinical trials partly due to their short serum half-lives. Our previous studies demonstrated that recombinant human plasminogen kringle 1-5 (K1-5) has superior antitumor activity compared with angiostatin. In addition, optimization of recombinant K1-5 with three amino acid substitutions enhances its antitumor effect. The current study was thus undertaken to evaluate prolonged expression of optimized K1-5 as cancer gene therapy. The recombinant adeno-associated virus (AAV) vector was used to express a secreted form of the optimized K1-5 (AAV-sK15tm) to improve its pharmacokinetic profile, which was considered to be the hurdle in angiostatin treatment of cancer. We successfully generated high-titer recombinant AAV vectors and observed sustained transgene expression for 567 days after a single injection of virus. The treated animals did not display any visible signs of abnormalities and showed normal serum biochemistry. The therapeutic potential of this treatment modality was demonstrated by both a strong inhibition of lung metastasis in the mouse B16F10 melanoma model and significant growth retardation of Lewis lung carcinoma xenografts in C57BL/6N mice as well as human A2058 melanoma xenografts in NOD/SCID (nonobese diabetic/severe combined immunodeficient) mice. Taken together, our results suggested that AAV-sK15tm produced long-term suppressive effects on cancer growth in vivo and should warrant serious consideration for clinical development.
authors.affiliation: 1 Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University , Tainan, Taiwan .
authors.forename: Cheng-Hsiang
authors.initials: CH
authors.lastname: Kuo
authors.affiliation: 1 Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University , Tainan, Taiwan .|3 Cardiovascular Research Center, National Cheng Kung University , Tainan, Taiwan .
authors.forename: Bi-Ing
authors.initials: BI
authors.lastname: Chang
authors.affiliation: 1 Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University , Tainan, Taiwan .|3 Cardiovascular Research Center, National Cheng Kung University , Tainan, Taiwan .
authors.forename: Fang-Tzu
authors.initials: FT
authors.lastname: Lee
authors.affiliation: 1 Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University , Tainan, Taiwan .
authors.forename: Po-Ku
authors.initials: PK
authors.lastname: Chen
authors.affiliation: 4 Harvard Gene Therapy Initiative, Harvard Medical School , Boston, Massachusetts.
authors.forename: Jeng-Shin
authors.initials: JS
authors.lastname: Lee
authors.affiliation: 1 Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University , Tainan, Taiwan .|2 Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University , Tainan, Taiwan .|3 Cardiovascular Research Center, National Cheng Kung University , Tainan, Taiwan .
authors.forename: Guey-Yueh
authors.initials: GY
authors.lastname: Shi
authors.affiliation: 1 Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University , Tainan, Taiwan .|2 Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University , Tainan, Taiwan .|3 Cardiovascular Research Center, National Cheng Kung University , Tainan, Taiwan .
authors.forename: Hua-Lin
authors.initials: HL
authors.lastname: Wu
mesh_terms: D000818:Animals; D017209:Apoptosis; D018827:Carcinoma, Lewis Lung / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics*; D015870:Gene Expression; D015316:Genetic Therapy; D057809:HEK293 Cells; D006801:Humans; D008175:Lung Neoplasms / Q000517:prevention & control* / Q000556:secondary; D008297:Male; D008545:Melanoma / Q000473:pathology / Q000628:therapy*; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic; D010958:Plasminogen / Q000235:genetics*; D017434:Protein Structure, Tertiary
chemical_list: D010958:Plasminogen