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.
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Abbreviations
- CMV:
-
cytomegalovirus
- EGFP:
-
enhanced green fluorescent protein
- HSV-1:
-
herpes simplex virus-1
- LDH:
-
lactate dehydrogenase
- MIS:
-
minimally invasive surgical
References
2004 Cancer Facts & Figures Available at: http://www.cancer.org/downloads/STT?CAFF_finalPWSecured.pdf (accessed July 1, 2004).
Carney DN . Lung cancer – time to move on from chemotherapy. N Engl J Med 2002; 346 (2): 126–128.
Roizman B . The function of herpes simplex virus genes: a primer for genetic engineering of novel vectors. Proc Natl Acad Sci USA 1996; 93 (21): 11307–11312.
Mineta T, Rabkin SD, Yazaki T, Hunter WD, Martuza RL . Attenuated multi-mutated herpes simplex virus-1 for the treatment of malignant gliomas. Nat Med 1995; 1 (9): 938–943.
Fong Y, Kemeny N, Jarnagin S, Stanziale S, Guilfoyle B, Gusani N et al. Phase 1 study of a replication-competent Herpes Simplex oncolytic virus for treatment of hepatic colorectal metastases. Abstract # 27. American Society of Clinical Oncology 38th Annual Meeting Proceedings, vol. 2: Part 1, 8a, 2002.
Rampling R, Cruickshank G, Papanastassiou V, Nicoll J, Hadley D, Brennan D et al. Toxicity evaluation of replication-competent herpes simplex virus (ICP 34.5 null mutant 1716) in patients with recurrent malignant glioma. Gene Therapy 2000; 7 (10): 859–866.
Carew JF, Kooby DA, Halterman MW, Federoff HJ, Fong Y . Selective infection and cytolysis of human head and neck squamous cell carcinoma with sparing of normal mucosa by a cytotoxic herpes simplex virus type 1 (G207). Hum Gene Ther 1999; 10 (10): 1599–1606.
Markert JM, Medlock MD, Rabkin SD, Gillespie GY, Todo T, Hunter WD et al. Conditionally replicating herpes simplex virus mutant, G207 for the treatment of malignant glioma: results of a phase I trial. Gene Therapy 2000; 7 (10): 867–874.
Stiles BM, Bhargava A, Adusumilli PS, Stanziale SF, Kim TH, Rusch VW et al. The replication-competent oncolytic herpes simplex mutant virus NV1066 is effective in the treatment of esophageal cancer. Surgery 2003; 134 (2): 357–364.
Bennett JJ, Adusumilli P, Petrowsky H, Burt BM, Roberts G, Delman KA et al. Up-regulation of GADD34 mediates the synergistic anticancer activity of mitomycin C and a gamma134.5 deleted oncolytic herpes virus (G207). FASEB J 2004; 18 (9): 1001–1003.
Jiao X, Krasna MJ . Clinical significance of micrometastasis in lung and esophageal cancer: a new paradigm in thoracic oncology. Ann Thorac Surg 2002; 74 (1): 278–284.
Dresler CM, Fratelli C, Babb J . Prognostic value of positive pleural lavage in patients with lung cancer resection. Ann Thorac Surg 1999; 67 (5): 1435–1439.
Okumura M, Ohshima S, Kotake Y, Morino H, Kikui M, Yasumitsu T . Intraoperative pleural lavage cytology in lung cancer patients. Ann Thorac Surg 1991; 51 (4): 599–604.
Higashiyama M, Doi O, Kodama K, Yokouchi H, Tateishi R, Horai T et al. Pleural lavage cytology immediately after thoracotomy and before closure of the thoracic cavity for lung cancer without pleural effusion and dissemination: clinicopathologic and prognostic analysis. Ann Surg Oncol 1997; 4 (5): 409–415.
Buhr J, Berghauser KH, Gonner S, Kelm C, Burkhardt EA, Padberg WM . The prognostic significance of tumor cell detection in intraoperative pleural lavage and lung tissue cultures for patients with lung cancer. J Thorac Cardiovasc Surg 1997; 113 (4): 683–690.
Kondo H, Asamura H, Suemasu K, Goya T, Tsuchiya R, Naruke T et al. Prognostic significance of pleural lavage cytology immediately after thoracotomy in patients with lung cancer. J Thorac Cardiovasc Surg 1993; 106 (6): 1092–1097.
Wong RJ, Joe JK, Kim SH, Shah JP, Horsburgh B, Fong Y . Oncolytic herpesvirus effectively treats murine squamous cell carcinoma and spreads by natural lymphatics to treat sites of lymphatic metastases. Hum Gene Ther 2002; 13 (10): 1213–1223.
Stanziale SF, Stiles BM, Bhargava A, Kerns SA, Kalakonda N, Fong Y . Oncolytic herpes simplex virus-1 mutant expressing green fluorescent protein can detect and treat peritoneal cancer. Hum Gene Ther 2004; 15 (6): 609–618.
Cormack BP, Valdivia RH, Falkow S . FACS-optimized mutants of the green fluorescent protein (GFP). Gene 1996; 173 (Spec. No.): 33–38.
Nunez R, Ackermann M, Saeki Y, Chiocca A, Fraefel C . Flow cytometric assessment of transduction efficiency and cytotoxicity of herpes simplex virus type 1-based amplicon vectors. Cytometry 2001; 44 (2): 93–99.
O'shea CC . DNA tumor viruses – the spies who lyse us. Curr Opin Genet Dev 2005; 15 (1): 18–26.
Toda M, Rabkin SD, Martuza RL . Treatment of human breast cancer in a brain metastatic model by G207, a replication-competent multimutated herpes simplex virus 1. Hum Gene Ther 1998; 9 (15): 2177–2185.
Nakamura H, Kasuya H, Mullen JT, Yoon SS, Pawlik TM, Chandrasekhar S et al. Regulation of herpes simplex virus gamma(1)34.5 expression and oncolysis of diffuse liver metastases by Myb34.5. J Clin Invest 2002; 109 (7): 871–882.
Ebright MI, Zager JS, Malhotra S, Delman KA, Weigel TL, Rusch VW et al. Replication-competent herpes virus NV1020 as direct treatment of pleural cancer in a rat model. J Thorac Cardiovasc Surg 2002; 124 (1): 123–129.
Kooby DA, Carew JF, Halterman MW, Mack JE, Bertino JR, Blumgart LH et al. Oncolytic viral therapy for human colorectal cancer and liver metastases using a multi-mutated herpes simplex virus type-1 (G207). FASEB J 1999; 13 (11): 1325–1334.
Varghese S, Rabkin SD . Oncolytic herpes simplex virus vectors for cancer virotherapy. Cancer Gene Ther 2002; 9 (12): 967–978.
Kjellberg SI, Dresler CM, Goldberg M . Pleural cytologies in lung cancer without pleural effusions. Ann Thorac Surg 1997; 64 (4): 941–944.
Okada M, Tsubota N, Yoshimura M, Miyamoto Y, Maniwa Y . Role of pleural lavage cytology before resection for primary lung carcinoma. Ann Surg 1999; 229 (4): 579–584.
Sahn SA, Good Jr JT . Pleural fluid pH in malignant effusions. Diagnostic, prognostic, and therapeutic implications. Ann Intern Med 1988; 108 (3): 345–349.
Johnston WW . The malignant pleural effusion. A review of cytopathologic diagnoses of 584 specimens from 472 consecutive patients. Cancer 1985; 56 (4): 905–909.
DiBonito L, Falconieri G, Colautti I, Bonifacio D, Dudine S . The positive pleural effusion. A retrospective study of cytopathologic diagnoses with autopsy confirmation. Acta Cytol 1992; 36 (3): 329–332.
Chernow B, Sahn SA . Carcinomatous involvement of the pleura: an analysis of 96 patients. Am J Med 1977; 63 (5): 695–702.
Ichinose Y, Tsuchiya R, Koike T, Yasumitsu T, Nakamura K, Tada H et al. A prematurely terminated phase III trial of intraoperative intrapleural hypotonic cisplatin treatment in patients with resected non-small cell lung cancer with positive pleural lavage cytology: the incidence of carcinomatous pleuritis after surgical intervention. J Thorac Cardiovasc Surg 2002; 123 (4): 695–699.
Jacobs A, Tjuvajev JG, Dubrovin M, Akhurst T, Balatoni J, Beattie B et al. Positron emission tomography-based imaging of transgene expression mediated by replication-conditional, oncolytic herpes simplex virus type 1 mutant vectors in vivo. Cancer Res 2001; 61 (7): 2983–2995.
Hemminki A, Zinn KR, Liu B, Chaudhuri TR, Desmond RA, Rogers BE et al. In vivo molecular chemotherapy and noninvasive imaging with an infectivity-enhanced adenovirus. J Natl Cancer Inst 2002; 94 (10): 741–749.
Lerondel S, Le Pape A, Sene C, Faure L, Bernard S, Diot P et al. Radioisotopic imaging allows optimization of adenovirus lung deposition for cystic fibrosis gene therapy. Hum Gene Ther 2001; 12: 1–11.
Bennett JJ, Tjuvajev J, Johnson P, Doubrovin M, Akhurst T, Malholtra S et al. Positron emission tomography imaging for herpes virus infection: implications for oncolytic viral treatments of cancer. Nat Med 2001; 7: 859–863.
Groot-Wassink T, Aboagye EO, Glaser M, Lemoine NR, Vassaux G . Adenovirus biodistribution and noninvasive imaging of gene expression in vivo by positron emission tomography using human sodium/iodide symporter as reporter gene. Hum Gene Ther 2002; 13 (14): 1723–1735.
Haberkorn U, Altmann A . Functional genomics and radioisotope-based imaging procedures. Ann Med 2003; 35 (6): 370–379.
Yang M, Baranov E, Jiang P, Sun FX, Li XM, Li L et al. Whole-body optical imaging of green fluorescent protein-expressing tumors and metastases. Proc Natl Acad Sci USA 2000; 97 (3): 1206–1211.
Yang M, Baranov E, Moossa AR, Penman S, Hoffman RM . Visualizing gene expression by whole-body fluorescence imaging. Proc Natl Acad Sci USA 2000; 97 (22): 12278–12282.
Jacobs A, Dubrovin M, Hewett J, Sena-Esteves M, Tan CW, Slack M et al. Functional coexpression of HSV-1 thymidine kinase and green fluorescent protein: implications for noninvasive imaging of transgene expression. Neoplasia 1999; 1 (2): 154–161.
Paquin A, Jaalouk DE, Galipeau J . Retrovector encoding a green fluorescent protein-herpes simplex virus thymidine kinase fusion protein serves as a versatile suicide/reporter for cell and gene therapy applications. Hum Gene Ther 2001; 12 (1): 13–23.
Mazurier F, Moreau-Gaudry F, Maguer-Satta V, Salesse S, Pigeonnier-Lagarde V, Ged C et al. Rapid analysis and efficient selection of human transduced primitive hematopoietic cells using the humanized S65T green fluorescent protein. Gene Therapy 1998; 5 (4): 556–562.
Tsutsudaasano A, Migita M, Takahashi K, Shimada T . Transduction of fibroblasts and CD34+ progenitors using a selectable retroviral vector containing cDNAs encoding arylsulfatase A and CD24. J Hum Genet 2000; 45 (1): 18–23.
Hunt L, Jordan M, De Jesus M, Wurm FM . GFP-expressing mammalian cells for fast, sensitive, noninvasive cell growth assessment in a kinetic mode. Biotechnol Bioeng 1999; 65 (2): 201–205.
Prosst RL, Winkler S, Boehm E, Gahlen J . Thoracoscopic fluorescence diagnosis (TFD) of pleural malignancies: experimental studies. Thorax 2002; 57 (12): 1005–1009.
Gahlen J, Stern J, Laubach HH, Pietschmann M, Herfarth C . Improving diagnostic staging laparoscopy using intraperitoneal lavage of delta-aminolevulinic acid (ALA) for laparoscopic fluorescence diagnosis. Surgery 1999; 126 (3): 469–473.
Gahlen J, Prosst RL, Pietschmann M, Rheinwald M, Haase T, Herfarth C . Spectrometry supports fluorescence staging laparoscopy after intraperitoneal aminolaevulinic acid lavage for gastrointestinal tumours. J Photochem Photobiol B 1999; 52 (1–3): 131–135.
Gahlen J, Prosst RL, Pietschmann M, Haase T, Rheinwald M, Skopp G et al. Laparoscopic fluorescence diagnosis for intraabdominal fluorescence targeting of peritoneal carcinosis experimental studies. Ann Surg 2002; 235 (2): 252–260.
Chishima T, Miyagi Y, Wang X, Yamaoka H, Shimada H, Moossa AR et al. Cancer invasion and micrometastasis visualized in live tissue by green fluorescent protein expression. Cancer Res 1997; 57 (10): 2042–2047.
Chishima T, Miyagi Y, Wang X, Baranov E, Tan Y, Shimada H et al. Metastatic patterns of lung cancer visualized live and in process by green fluorescence protein expression. Clin Exp Metastasis 1997; 15 (5): 547–552.
Chishima T, Miyagi Y, Wang X, Tan Y, Shimada H, Moossa A et al. Visualization of the metastatic process by green fluorescent protein expression. Anticancer Res 1997; 17 (4A): 2377–2384.
Hasegawa S, Yang M, Chishima T, Miyagi Y, Shimada H, Moossa AR et al. In vivo tumor delivery of the green fluorescent protein gene to report future occurrence of metastasis. Cancer Gene Ther 2000; 7 (10): 1336–1340.
Diehn FE, Costouros NG, Miller MS, Feldman AL, Alexander HR, Li KC et al. Noninvasive fluorescent imaging reliably estimates biomass in vivo. Biotechniques 2002; 33 (6): 1250–1255.
Robinson PJ . Imaging liver metastases: current limitations and future prospects. Br J Radiol 2000; 73 (867): 234–241.
Hirsch FR, Franklin WA, Gazdar AF, Bunn Jr PA . Early detection of lung cancer: clinical perspectives of recent advances in biology and radiology. Clin Cancer Res 2001; 7 (1): 5–22.
Acknowledgements
We thank Brian Horsburgh, PhD and Medigene, Inc. for constructing and providing us with the NV1066 virus. We thank Mithat Gonen, PhD, of the Department of Epidemiology and Biostatistics, and Katia Manova, PhD, John Waka, BS, and members of the Molecular Cytology Core Facility at Memorial Sloan-Kettering Cancer Center for their assistance with this project. We thank Hai Nguyen, VMD, of the Research Animal Resource Center of Cornell University Medical College for review of pathology specimens. Special thanks to Kan Matsumoto from Olympus America Inc., for design and construction of the fluorescent thoracoscopic system. This study was presented at the sixth annual meeting of the ‘American Society of Gene Therapy’, 2003, in Washington DC. This work was supported in part by training grant T32 CA09501 (BMS), AACR-AstraZeneca Cancer Research and Prevention Foundation Fellowship (PSA), grants RO1 CA 76416 and RO1 CA/DK80982 (YF) from the National Institutes of Health, grant MBC-99366 (YF) from the American Cancer Society, grant BC024118 from the US Army (YF), grant IMG0402501 from the Susan Komen Cancer foundation (YF and PSA) and grant 032047 from Flight Attendant Medical Research Institute (YF and PSA).
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Stiles, B., Adusumilli, P., Bhargava, A. et al. Minimally invasive localization of oncolytic herpes simplex viral therapy of metastatic pleural cancer. Cancer Gene Ther 13, 53–64 (2006). https://doi.org/10.1038/sj.cgt.7700860
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DOI: https://doi.org/10.1038/sj.cgt.7700860
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