title: Minimally invasive localization of oncolytic herpes simplex viral therapy of metastatic pleural cancer.
abstract: Herpes simplex virus-1 (HSV-1) oncolytic therapy and gene therapy are promising treatment modalities against cancer. NV1066, one such HSV-1 virus, carries a marker gene for enhanced green fluorescent protein (EGFP). The purpose of this study was to determine whether NV1066 is cytotoxic to lung cancer and whether EGFP is a detectable marker of viral infection in vitro and in vivo. We further investigated whether EGFP expression in infected cells can be used to localize the virus and to identify small metastatic tumor foci (<1 mm) in vivo by means of minimally invasive endoscopic systems equipped with fluorescent filters. In A549 human lung cancer cells, in vitro viral replication was determined by plaque assay, cell kill by LDH release assay, and EGFP expression by flow cytometry. In vivo, A549 cells were injected into the pleural cavity of athymic mice. Mice were treated with intrapleural injection of NV1066 or saline and examined for EGFP expression in tumor deposits using a stereomicroscope or a fluorescent thoracoscopic system. NV1066 replicated in, expressed EGFP in infected cells and killed tumor cells in vitro. In vivo, treatment with intrapleural NV1066 decreased pleural disease burden, as measured by chest wall nodule counts and organ weights. EGFP was easily visualized in tumor deposits, including microscopic foci, by fluorescent thoracoscopy. NV1066 has significant oncolytic activity against a human NSCLC cell line and is effective in limiting the progression of metastatic disease in an in vivo orthotopic model. By incorporating fluorescent filters into endoscopic systems, a minimally invasive means for diagnosing small metastatic pleural deposits and localization of viral therapy for thoracic malignancies may be developed using the EGFP marker gene inserted in oncolytic herpes simplex viruses.
authors.affiliation: Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
authors.forename: B M
authors.initials: BM
authors.lastname: Stiles
authors.forename: P S
authors.initials: PS
authors.lastname: Adusumilli
authors.forename: A
authors.initials: A
authors.lastname: Bhargava
authors.forename: S F
authors.initials: SF
authors.lastname: Stanziale
authors.forename: T H
authors.initials: TH
authors.lastname: Kim
authors.forename: M-K
authors.initials: MK
authors.lastname: Chan
authors.forename: R
authors.initials: R
authors.lastname: Huq
authors.forename: R
authors.initials: R
authors.lastname: Wong
authors.forename: V W
authors.initials: VW
authors.lastname: Rusch
authors.forename: Y
authors.initials: Y
authors.lastname: Fong
mesh_terms: D000818:Animals; D005434:Flow Cytometry; D005470:Fluorometry; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D010997:Pleural Neoplasms / Q000473:pathology / Q000628:therapy* / Q000821:virology*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014407:Tumor Cells, Cultured
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins