Bacterial glucuronidase as general marker for oncolytic virotherapy or other biological therapies

Journal of Translational Medicine

Aladar A Szalay²  Ivaylo Gentschev³  Julia B Sturm⁴  Barbara Härtl¹  Jochen Stritzker³  Michael Hess⁴ 

[1] Genelux GmbH, Bernried, Germany;Department of Radiation Oncology, Moores Cancer Center, University of California, San Diego, La Jolla, CA, USA;Genelux Corporation, San Diego, CA, USA;Department of Biochemistry, Biocenter, University of Würzburg, Würzburg, Germany

关键词: fluorescent probe;    reporter;    cancer;    oncolytic virus;    beta-glucuronidase;   

Others  :  1207846
DOI  :  10.1186/1479-5876-9-172

 received in 2011-08-08, accepted in 2011-10-11,  发布年份 2011

【 摘 要 】

Background

Oncolytic viral tumor therapy is an emerging field in the fight against cancer with rising numbers of clinical trials and the first clinically approved product (Adenovirus for the treatment of Head and Neck Cancer in China) in this field. Yet, until recently no general (bio)marker or reporter gene was described that could be used to evaluate successful tumor colonization and/or transgene expression in other biological therapies.

Methods

Here, a bacterial glucuronidase (GusA) encoded by biological therapeutics (e.g. oncolytic viruses) was used as reporter system.

Results

Using fluorogenic probes that were specifically activated by glucuronidase we could show 1) preferential activation in tumors, 2) renal excretion of the activated fluorescent compounds and 3) reproducible detection of GusA in the serum of oncolytic vaccinia virus treated, tumor bearing mice in several tumor models. Time course studies revealed that reliable differentiation between tumor bearing and healthy mice can be done as early as 9 days post injection of the virus. Regarding the sensitivity of the newly developed assay system, we could show that a single infected tumor cell could be reliably detected in this assay.

Conclusion

GusA therefore has the potential to be used as a general marker in the preclinical and clinical evaluation of (novel) biological therapies as well as being useful for the detection of rare cells such as circulating tumor cells.

【 授权许可】

   
2011 Hess et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Kirn DH, Thorne SH: Targeted and armed oncolytic poxviruses: a novel multi-mechanistic therapeutic class for cancer. Nat Rev Cancer 2009, 9:64-71.
• [2]Garcia-Aragoncillo E, Hernandez-Alcoceba R: Design of virotherapy for effective tumor treatment. Curr Opin Mol Ther 2010, 12:403-411.
• [3]Moss B: Genetically engineered poxviruses for recombinant gene expression, vaccination, and safety. Proc Natl Acad Sci USA 1996, 93:11341-11348.
• [4]Zhang Q, Yu YA, Wang E, Chen N, Danner RL, Munson PJ, Marincola FM, Szalay AA: Eradication of solid human breast tumors in nude mice with an intravenously injected light-emitting oncolytic vaccinia virus. Cancer Res 2007, 67:10038-10046.
• [5]Yu YA, Shabahang S, Timiryasova TM, Zhang Q, Beltz R, Gentschev I, Goebel W, Szalay AA: Visualization of tumors and metastases in live animals with bacteria and vaccinia virus encoding light-emitting proteins. Nat Biotechnol 2004, 22:313-320.
• [6]Puhlmann M, Brown CK, Gnant M, Huang J, Libutti SK, Alexander HR, Bartlett DL: Vaccinia as a vector for tumor-directed gene therapy: biodistribution of a thymidine kinase-deleted mutant. Cancer Gene Ther 2000, 7:66-73.
• [7]Gentschev I, Stritzker J, Hofmann E, Weibel S, Yu YA, Chen N, Zhang Q, Bullerdiek J, Nolte I, Szalay AA: Use of an oncolytic vaccinia virus for the treatment of canine breast cancer in nude mice: preclinical development of a therapeutic agent. Cancer Gene Ther 2009, 16:320-328.
• [8]Heo J, Breitbach CJ, Moon A, Kim CW, Patt R, Kim MK, Lee YK, Oh SY, Woo HY, Parato K, et al.: Sequential Therapy With JX-594, A Targeted Oncolytic Poxvirus, Followed by Sorafenib in Hepatocellular Carcinoma: Preclinical and Clinical Demonstration of Combination Efficacy. Mol Ther 2011, 19:1170-1179.
• [9]Liu TC, Hwang T, Park BH, Bell J, Kirn DH: The targeted oncolytic poxvirus JX-594 demonstrates antitumoral, antivascular, and anti-HBV activities in patients with hepatocellular carcinoma. Mol Ther 2008, 16:1637-1642.
• [10]Park BH, Hwang T, Liu TC, Sze DY, Kim JS, Kwon HC, Oh SY, Han SY, Yoon JH, Hong SH, et al.: Use of a targeted oncolytic poxvirus, JX-594, in patients with refractory primary or metastatic liver cancer: a phase I trial. Lancet Oncol 2008, 9:533-542.
• [11]Pedersen J, Karapanagiotou L, Alam S, Puglisi M, Britton L, Sassi S, Mansfield D, Yap T, De-Bono J, Harrington K: Preliminary results of a Phase 1 study of intravenous administration of GL-ONC1 Vaccinia virus in patients with advanced solid cancer with real time imaging. 6th NCRI Cancer Conference; BT Convention Center, Liverpool, UK; 2010.
• [12]Bennett JJ, Tjuvajev J, Johnson P, Doubrovin M, Akhurst T, Malholtra S, Hackman T, Balatoni J, Finn R, Larson SM, et al.: Positron emission tomography imaging for herpes virus infection: Implications for oncolytic viral treatments of cancer. Nat Med 2001, 7:859-863.
• [13]Chen N, Zhang Q, Yu YA, Stritzker J, Brader P, Schirbel A, Samnick S, Serganova I, Blasberg R, Fong Y, Szalay AA: A novel recombinant vaccinia virus expressing the human norepinephrine transporter retains oncolytic potential and facilitates deep-tissue imaging. Mol Med 2009, 15:144-151.
• [14]Dingli D, Peng KW, Harvey ME, Greipp PR, O'Connor MK, Cattaneo R, Morris JC, Russell SJ: Image-guided radiovirotherapy for multiple myeloma using a recombinant measles virus expressing the thyroidal sodium iodide symporter. Blood 2004, 103:1641-1646.
• [15]Haddad D, Chen NG, Zhang Q, Chen CH, Yu YA, Gonzalez L, Carpenter SG, Carson J, Au J, Mittra A, et al.: Insertion of the human sodium iodide symporter to facilitate deep tissue imaging does not alter oncolytic or replication capability of a novel vaccinia virus. J Transl Med 2011, 9:36. BioMed Central Full Text
• [16]Fang W, Vikerpuur M, Sandholm M: A fluorometric beta-glucuronidase assay for analysis of bacterial growth in milk. Vet Microbiol 1995, 46:361-367.
• [17]Jefferson RA, Burgess SM, Hirsh D: beta-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci USA 1986, 83:8447-8451.
• [18]Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 1987, 6:3901-3907.
• [19]Stahl P, Fishman W: Beta-D-glucuronidase. In Methods in enzymatic analysis. Edited by JB, MG. Weinheim, Germany: Verlag Chemie; 1984:246-256.
• [20]Wang SM, Chern JW, Yeh MY, Ng JC, Tung E, Roffler SR: Specific activation of glucuronide prodrugs by antibody-targeted enzyme conjugates for cancer therapy. Cancer Res 1992, 52:4484-4491.
• [21]Cheng CM, Lu YL, Chuang KH, Hung WC, Shiea J, Su YC, Kao CH, Chen BM, Roffler S, Cheng TL: Tumor-targeting prodrug-activating bacteria for cancer therapy. Cancer Gene Ther 2008, 15:393-401.
• [22]de Graaf M, Pinedo HM, Oosterhoff D, van der Meulen-Muileman IH, Gerritsen WR, Haisma HJ, Boven E: Pronounced antitumor efficacy by extracellular activation of a doxorubicin-glucuronide prodrug after adenoviral vector-mediated expression of a human antibody-enzyme fusion protein. Hum Gene Ther 2004, 15:229-238.
• [23]Huang PT, Chen KC, Prijovich ZM, Cheng TL, Leu YL, Roffler SR: Enhancement of CPT-11 antitumor activity by adenovirus-mediated expression of beta-glucuronidase in tumors. Cancer Gene Ther 2011, 18:381-389.
• [24]Tzou SC, Roffler S, Chuang KH, Yeh HP, Kao CH, Su YC, Cheng CM, Tseng WL, Shiea J, Harm IH, et al.: Micro-PET imaging of beta-glucuronidase activity by the hydrophobic conversion of a glucuronide probe. Radiology 2009, 252:754-762.
• [25]Antunes IF, Haisma HJ, Elsinga PH, Dierckx RA, de Vries EF: Synthesis and evaluation of [18F]-FEAnGA as a PET Tracer for beta-glucuronidase activity. Bioconjug Chem 2010, 21:911-920.
• [26]Su YC, Chuang KH, Wang YM, Cheng CM, Lin SR, Wang JY, Hwang JJ, Chen BM, Chen KC, Roffler S, Cheng TL: Gene expression imaging by enzymatic catalysis of a fluorescent probe via membrane-anchored beta-glucuronidase. Gene Ther 2007, 14:565-574.
• [27]Sterenczak KA, Willenbrock S, Barann M, Klemke M, Soller JT, Eberle N, Nolte I, Bullerdiek J, Murua Escobar H: Cloning, characterisation, and comparative quantitative expression analyses of receptor for advanced glycation end products (RAGE) transcript forms. Gene 2009, 434:35-42.
• [28]Stritzker J, Weibel S, Hill PJ, Oelschlaeger TA, Goebel W, Szalay AA: Tumor-specific colonization, tissue distribution, and gene induction by probiotic Escherichia coli Nissle 1917 in live mice. Int J Med Microbiol 2007, 297:151-162.
• [29]Seubert CM, Stritzker J, Hess M, Donat U, Sturm JB, Chen N, von Hof JM, Krewer B, Tietze LF, Gentschev I, Szalay AA: Enhanced tumor therapy using vaccinia virus strain GLV-1h68 in combination with a beta-galactosidase-activatable prodrug seco-analog of duocarmycin SA. Cancer Gene Ther 2011, 18:42-52.
• [30]Chen KC, Wu CH, Chang CY, Lu WC, Tseng Q, Prijovich ZM, Schechinger W, Liaw YC, Leu YL, Roffler SR: Directed evolution of a lysosomal enzyme with enhanced activity at neutral pH by mammalian cell-surface display. Chem Biol 2008, 15:1277-1286.
• [31]Sharma SK, Bagshawe KD, Melton RG, Sherwood RF: Human immune response to monoclonal antibody-enzyme conjugates in ADEPT pilot clinical trial. Cell Biophys 1992, 21:109-120.
• [32]Chen KC, Wu SY, Leu YL, Prijovich ZM, Chen BM, Wang HE, Cheng TL, Roffler SR: A humanized immunoenzyme with enhanced activity for glucuronide prodrug activation in the tumor microenvironment. Bioconjug Chem 2011, 22:938-948.
• [33]Heine D, Muller R, Brusselbach S: Cell surface display of a lysosomal enzyme for extracellular gene-directed enzyme prodrug therapy. Gene Ther 2001, 8:1005-1010.
• [34]Weyel D, Sedlacek HH, Muller R, Brusselbach S: Secreted human beta-glucuronidase: a novel tool for gene-directed enzyme prodrug therapy. Gene Ther 2000, 7:224-231.