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Wo 2008/156655 A9 (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) CORRECTED VERSION (19) World Intellectual Property Organization International Bureau (43) International Publication Date (10) International Publication Number 24 December 2008 (24.12.2008) PCT WO 2008/156655 A9 (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every C07K 14/00 (2006.01) kind of national protection available): AE, AG, AL, AM, AO, AT,AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, CA, (21) International Application Number: CH, CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, EC, EE, PCT/US2008/007377 EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC, (22) International Filing Date: 13 June 2008 (13.06.2008) LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PG, PH, (25) Filing Language: English PL, PT, RO, RS, RU, SC, SD, SE, SG, SK, SL, SM, SV, SY, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, (26) Publication Language: English ZA, ZM, ZW (30) Priority Data: (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, 60/934,768 15 June 2007 (15.06.2007) US GM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), (71) Applicants (for all designated States except US): European (AT,BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, GENELUX CORPORATION [US/US]; 3030 Bunker FR, GB, GR, HR, HU, IE, IS, IT, LT,LU, LV,MC, MT, NL, Hill Street, Suite 310, San Diego, CA 92109 (US). NO, PL, PT, RO, SE, SI, SK, TR), OAPI (BF, BJ, CF, CG, SLOAN KETTERING INSTITUTE FOR CANCER CI, CM, GA, GN, GQ, GW, ML, MR, NE, SN, TD, TG). RESEARCH [US/US]; 1275 York Avenue, New York, NY 10021 (US). Declarations under Rule 4.17: — as to applicant's entitlement to applyfor and be granted a (72) Inventors; and patent (Rule 4.17(U)) (75) US only): Inventors/Applicants (for FONG, Yuman — as to the applicant's entitlement to claim the priority of the [US/US]; 345 East 68th Street, Apt., #4B, New York, NY earlier application (Rule 4.17(Ui)) 10021 (US). CHEN, Nanhai [CN/US]; 9167 Buckwheat Street, San Diego, CA 92129 (US). ZHANG, Qian Published: [CN/US]; 12601 E. Camino Real, Unit B, San Diego, CA — without international search report and to be republished 92130 (US). YU, Yong,A. [CN/US]; 3830 Elijah Ct., Unit upon receipt of that report 422, San Diego, CA 92130 (US). SZALAY, Aladar, A. [US/US]; 7704 North Fork Rd., Highland, CA 92346 (US). (48) Date of publication of this corrected version: 19 March 2009 (74) Agents: SEIDMAN, Stephanie, L. et al; Bell Boyd & Lloyd LLP, 3580 Carmel Mountain Road, Suite 200, San (15) Information about Correction: Diego, CA 92130 (US). see Notice of 19 March 2009 (54) Title: MICROORGANISMS FOR IMAGING AND/OR TREATMENT OF TUMORS (57) Abstract: Modified viruses encoding transporter proteins and methods for preparing the modified viruses are provided. Vac cines that contain the viruses are provided. The viruses also can be used in diagnostic methods, such detection and imaging of tumors. The viruses also can be used in methods of treatment of diseases, such as proliferative and inflammatory disorders, includ ing as anti-tumor agents. MICROORGANISMS FOR IMAGING AND/OR TREATMENT OF TUMORS RELATED APPLICATIONS Benefit of priority is claimed to U.S. Provisional Application Serial No. 60/934,768, to Nanhai Chen, Yuman Fong, Aladar A. Szalay, Yong A. Yu and Qian Zhang, filed on June 15, 2007, entitled "MICROORGANISMS FOR IMAGING AND/OR TREATMENT OF TUMORS." Where permitted, the subject matter of this application is incorporated by reference in its entirety. This application is related to U.S. Application Serial No. 12/157,960 to Nanhai Chen, Aladar A. Szalay, Yong A. Yu, Qian Zhang and Yuman Fong, filed on June 13, 2008, entitled "MICROORGANISMS FOR IMAGING AND/OR TREATMENT OF TUMORS," which also claims priority to U.S. Provisional Application Serial No. 60/934,768. Where permitted, the subject matter of this application is incorporated by reference in its entirety. FIELD OF THE INVENTION Modified recombinant viruses for diagnosis and therapy are provided. Diagnostic and therapeutic methods using the modified recombinant viruses also are provided. BACKGROUND Cancers, such as pancreatic cancer and malignant pleural mesothelioma, are highly aggressive diseases. The annual incidence in the United States was estimated to be ~40,000 cases for pancreatic cancer and -4,000 cases for malignant mesothelioma in the year 2004, with increasing incidence worldwide for mesothelioma, especially in industrialized nations due to the etiology of this disease from asbestos exposure (Bianchi and Bianchi (2007) Ind Health 45: 379-87). Both of these tumors are highly resistant to standard therapies, with 5-year survival rates of only 5% for pancreatic cancer and 9% for mesothelioma. Even with combined surgery, chemotherapy and radiation, only a small minority of patients are rendered disease-free for a prolonged period of time (Adusumilli et al. (2006) J Gene Med 8:603-1 5. Oncolytic viral therapy has been studied and tested over the past century, and many viral types, including adenovirus, herpes simplex virus, Newcastle disease virus, myxoma virus, vaccinia virus and vesicular stomatitis virus, are being investigated as novel agents for the treatment of human cancer (Woo et al (2006) Curr Opin Investig Drugs 7:549-59). Accordingly, effective tumor diagnostic and therapeutic viral agents that are highly selective for tumors are needed. In addition, there exists a need to provide reagents and methods for tracking and monitoring viral distribution, tumor targeting, proliferation and persistence in oncolytic viral therapies by noninvasive imaging, which provide important safety, efficacy and toxicity data. Such real-time monitoring also would provide useful viral-dose and administration schedule information for optimization of therapy and would obviate the need for multiple and repeated tissue biopsies. SUMMARY Provided are recombinant viruses, particularly, vaccinia virus, such as LIVP, that accumulates in tumors or other immunoprivileged tissues, such as wounds and inflamed tissues, and not accumulate to toxic levels in other tissues. These viruses encoded a protein that enhances uptake or retention of a compound that emits a signal that permits detection, such as by non-optical imaging. Proteins that enhance uptake or retention include transporter proteins. These viruses also can be used for treatment of tumors, wounded tissues and inflammations within a subject. The compound that is taken up or retained can be a therapeutic compound or can be modified, such as by conjugate to a therapeutic compound, to have therapeutic activity. The viruses can be used for detection, detection and treatment, detection and monitoring of treatment. Methods for detection, detection and treatment, detection and monitoring of treatment are provided as are uses of recombinant viruses, such as vaccinia viruses for detection, detection and treatment, detection and monitoring of treatment. Provided herein are recombinant vaccinia viruses that encode a sodium-dependent transporter protein. Sodium-dependent transporter proteins include those from the solute carrier 5 and solute carrier 6 transporter protein families, such as a norepinephrine transporter (NET) and a sodium-iodide symporter (NIS), including a human norepinephrine transporter (hNET) and a human sodium-iodide symporter (hNIS) as well as allelic and species variants thereof and other variants, including any having at least about or at least 60, 65, 70, 75, 80, 85, 88, 90, 91, 92, 93, 94, 95, 96, 97, 98 and 99 percent or more sequence identity with those disclosed herein. These include modified forms that retain transporter activity sufficient for the methods provided herein. Recombinant vaccinia virus include of any of claims 1-5 that is a Lister strain viruses, such as the LIVP strain. Nucleic acid encoding transporter protein can be inserted anywhere in the virus such that the virus expresses it and replicates in a subject. In exemplary embodiments, the nucleic acid encoding the transporter protein is inserted into a nonessential locus or gene, such as the hemagglutinin (HA), thymidine kinase (TK) or Fl4.5 gene or locus. Exemplary of such viruses are those provided herein that include GLV-lh99, GLV-IhIOO, GLV-IhIOl, GLV-1M39, GLV-lhl46, GLV-lhl50, GLV- IhI 51, GLV-lhl52 and GLV-lhl53. These viruses can be further modified to encode a therapeutic protein. Generally the encoding nucleic acid is inserted into a different locus from the nucleic acid that encodes the transporter protein. Exemplary therapeutic agents include, but are not limited to an anti-cancer agents and anti-angiogenic agents. A therapeutic agent, includes, but is not limited to, a cytokine, a chemokine, an immunomodulatory molecule, an antigen, an antibody or fragment thereof, antisense RNA, prodrug converting enzyme, siRNA, angiogenesis inhibitor, a toxin, an antitumor oligopeptide* a mitosis inhibitor protein, an antimitotic oligopeptide, an anti-cancer polypeptide antibiotic, and tissue factor, such as single chain antibody (scFv), including an anti-VEGF single chain antibody, a plasminogen K5 domain, a human tissue factor- αv/33-integrin RGD fusion protein, interleukin-24 or an IL-6-IL-6 receptor fusion protein and fusion proteins of substrates for the transporter protein and a therapeutic agent, such as a chemotherapeutic compound or a toxin. The viruses can encode a plurality of therapeutic agents and/or transporter proteins. Also provided are combination that contain one or more of the recombinant viruses, particularly vaccinia viruses, provided herein, and a substrate transported into a cell that expresses the transporter; and/or an anti-cancer compound.
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