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. Access through your

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DRIVEN BY THE PSA PROMOTER Article Open access 11 December 2023 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. Article  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS

  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS 

Google Scholar  * 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. Article  Google Scholar  * 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. Article  CAS  Google Scholar  *

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. Article  Google

Scholar  * 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. Article  CAS 

Google Scholar  * 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.

Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  *

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. Article  CAS  Google Scholar  * 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. CAS  PubMed  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * Cormack BP, Valdivia RH, Falkow S .

FACS-optimized mutants of the green fluorescent protein (GFP). _Gene_ 1996; 173 (Spec. No.): 33–38. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  *

O'shea CC . DNA tumor viruses – the spies who lyse us. _Curr Opin Genet Dev_ 2005; 15 (1): 18–26. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  *

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. Article  CAS  Google Scholar  * 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. Article  Google Scholar  * 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.

Article  CAS  Google Scholar  * Varghese S, Rabkin SD . Oncolytic herpes simplex virus vectors for cancer virotherapy. _Cancer Gene Ther_ 2002; 9 (12): 967–978. Article  CAS  Google Scholar

  * Kjellberg SI, Dresler CM, Goldberg M . Pleural cytologies in lung cancer without pleural effusions. _Ann Thorac Surg_ 1997; 64 (4): 941–944. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  *

Sahn SA, Good Jr JT . Pleural fluid pH in malignant effusions. Diagnostic, prognostic, and therapeutic implications. _Ann Intern Med_ 1988; 108 (3): 345–349. Article  CAS  Google Scholar  *

Johnston WW . The malignant pleural effusion. A review of cytopathologic diagnoses of 584 specimens from 472 consecutive patients. _Cancer_ 1985; 56 (4): 905–909. Article  CAS  Google

Scholar  * 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. CAS  PubMed  Google Scholar  * Chernow B, Sahn SA . Carcinomatous involvement of the pleura: an analysis of 96 patients. _Am J Med_ 1977; 63 (5): 695–702.

Article  CAS  Google Scholar  * 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. Article  Google Scholar  * 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. CAS  Google Scholar

  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  *

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. Article  CAS  Google Scholar  * Haberkorn U, Altmann A . Functional genomics and radioisotope-based

imaging procedures. _Ann Med_ 2003; 35 (6): 370–379. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * Prosst

RL, Winkler S, Boehm E, Gahlen J . Thoracoscopic fluorescence diagnosis (TFD) of pleural malignancies: experimental studies. _Thorax_ 2002; 57 (12): 1005–1009. Article  CAS  Google Scholar 

* 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google

Scholar  * 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. Article  Google Scholar  * 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. CAS  Google Scholar  * 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. Article  CAS  Google Scholar  *

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.

CAS  PubMed  Google Scholar  * 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. Article  CAS  Google Scholar  * 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. Article  CAS  Google Scholar  * Robinson PJ . Imaging liver metastases: current limitations

and future prospects. _Br J Radiol_ 2000; 73 (867): 234–241. Article  CAS  Google Scholar  * 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. CAS  PubMed  Google Scholar  Download references 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). AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

B M Stiles, P S Adusumilli, A Bhargava, S F Stanziale, T H Kim, M-K Chan, R Wong, V W Rusch & Y Fong * Molecular Cytology Core Facility, Memorial Sloan-Kettering Cancer Center, New York,

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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 Download citation * Received: 09 November 2004 * Revised: 16 March 2005 * Accepted: 17 March 2005 * Published: 22 July 2005 * Issue Date: 01 January

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is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * green fluorescent protein * lung neoplasm *

minimally invasive * targeted therapy * thoracoscopy * bronchoscopy