title: A targeting ligand enhances infectivity and cytotoxicity of an oncolytic adenovirus in human pancreatic cancer tissues.
abstract: The addition of a targeting strategy is necessary to enhance oncolysis and secure safety of a conditionally replicative adenovirus (CRAd). We have constructed an adenovirus library displaying random peptides on the fiber, and have successfully identified a pancreatic cancer-targeting ligand (SYENFSA). Here, the usefulness of cancer-targeted CRAd for pancreatic cancer was examined as a preclinical study. First, we constructed a survivin promoter-regulated CRAd expressing enhanced green fluorescent protein gene (EGFP), which displayed the identified targeting ligand (AdSur-SYE). The AdSur-SYE resulted in higher gene transduction efficiency and oncolytic potency than the untargeted CRAd (AdSur) in several pancreatic cancer cell lines. An intratumoral injection of AdSur-SYE significantly suppressed the growth of subcutaneous tumors, in which AdSur-SYE effectively proliferated and spread. An ectopic infection in adjacent tissues and organs of intratumorally injected AdSur-SYE was decreased compared with AdSur. Then, to examine whether the targeting ligand actually enhanced the infectivity of CRAd in human pancreatic cancer tissues, tumor cells prepared from surgical specimens were infected with viruses. The AdSur-SYE increased gene transduction efficiency 6.4-fold higher than did AdSur in single cells derived from human pancreatic cancer, whereas the infectivity of both vectors was almost the same in the pancreas and other cancers. Immunostaining showed that most EGFP(+) cells were cytokeratin-positive in the sliced tissues, indicating that pancreatic cancer cells but not stromal cells were injected with AdSur-SYE. AdSur-SYE resulted in a stronger oncolysis in the primary pancreatic cancer cells co-cultured with mouse embryonic fibroblasts than AdSur did. CRAd in combination with a tumor-targeting ligand is promising as a next-generation of oncolytic virotherapy for pancreatic cancer.
authors.affiliation: Division of Gene and Immune Medicine, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan; Laboratory of Oncology, Tokyo University of Pharmacy and Life Sciences, 432-1 Horinouchi, Hachioji, Tokyo 192-0355, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yamamoto
authors.affiliation: Division of Molecular Pathology, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
authors.forename: Nobuyoshi
authors.initials: N
authors.lastname: Hiraoka
authors.affiliation: Division of Gene and Immune Medicine, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Goto
authors.affiliation: Division of Gene and Immune Medicine, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan; Laboratory of Oncology, Tokyo University of Pharmacy and Life Sciences, 432-1 Horinouchi, Hachioji, Tokyo 192-0355, Japan.
authors.forename: Yosei
authors.initials: Y
authors.lastname: Rin
authors.affiliation: Division of Gene and Immune Medicine, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan; Laboratory of Oncology, Tokyo University of Pharmacy and Life Sciences, 432-1 Horinouchi, Hachioji, Tokyo 192-0355, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Miura
authors.affiliation: Division of Gene and Immune Medicine, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
authors.forename: Kenta
authors.initials: K
authors.lastname: Narumi
authors.affiliation: Laboratory of Oncology, Tokyo University of Pharmacy and Life Sciences, 432-1 Horinouchi, Hachioji, Tokyo 192-0355, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Uchida
authors.affiliation: Division of Pathology and Cell Therapy, Chiba Cancer Center Research Institute, 666-2 Nitona-chou, Chuo-ku, Chiba 260-0801, Japan.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Tagawa
authors.affiliation: Division of Gene and Immune Medicine, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan. Electronic address: kaoki@ncc.go.jp.
authors.forename: Kazunori
authors.initials: K
authors.lastname: Aoki
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D050130:Oncolytic Virotherapy / Q000379:methods*; D050504:Oncolytic Viruses / Q000235:genetics*; D010179:Pancreas / Q000378:metabolism / Q000473:pathology; D010190:Pancreatic Neoplasms / Q000473:pathology / Q000628:therapy*; D011401:Promoter Regions, Genetic; D000077022:Survivin
chemical_list: C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D000077022:Survivin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Oncolytic adenovirus; Pancreatic cancer; Survivin promoter; Targeting