Microorganisms for imaging and/or treatment of tumors

Abstract

Modified viruses encoding transporter proteins and methods for preparing the modified viruses are provided. Vaccines 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, including as anti-tumor agents.

1 . A recombinant vaccinia virus that encodes a sodium-dependent transporter protein. 
     
     
         2 . The recombinant vaccinia virus of  claim 1 , wherein the transporter protein is selected from among a solute carrier 5 family and solute carrier 6 transporter protein. 
     
     
         3 . The recombinant vaccinia virus of  claim 1 , wherein the transporter protein is a norepinephrine transporter (NET) or is a sodium-iodide symporter (NIS). 
     
     
         4 . The recombinant vaccinia virus of  claim 3 , wherein the transporter protein is a norepinephrine transporter that is a human norepinephrine transporter (hNET). 
     
     
         5 . The recombinant vaccinia virus of  claim 3 , wherein the transporter protein is a sodium-iodide symporter that is a human sodium-iodide symporter (hNIS). 
     
     
         6 . The recombinant vaccinia virus of  claim 1 , that is a Lister strain virus. 
     
     
         7 . The recombinant vaccinia virus of  claim 6  that is an LIVP strain virus. 
     
     
         8 . The recombinant vaccinia virus of  claim 7  that is selected from among GLV-1h99, GLV-1h100, GLV-1h101, GLV-1h139, GLV-1h146, GLV-1h150, GLV-1h151, GLV-1h152 and GLV-1h153. 
     
     
         9 . The recombinant vaccinia virus of  claim 1 , wherein nucleic acid encoding the transporter protein is inserted in the hemagglutinin (HA), thymidine kinase (TK) or F14.5 gene or locus. 
     
     
         10 . The recombinant vaccinia virus of  claim 1 , wherein the virus encodes a therapeutic agent. 
     
     
         11 . The recombinant vaccinia virus of  claim 10 , wherein the therapeutic agent is an anti-cancer agent or anti-angiogenic agent. 
     
     
         12 . The recombinant vaccinia virus of  claim 10 , wherein the therapeutic agent is inserted into a different gene or locus from the transporter gene and is inserted in the hemagglutinin (HA), thymidine kinase (TK) or F14.5 gene or locus. 
     
     
         13 . The recombinant vaccinia virus of  claim 10 , wherein the therapeutic agent is selected from among 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. 
     
     
         14 . The recombinant virus of  claim 13  wherein the therapeutic agent is an antibody or fragment thereof that is a single chain antibody (scFv). 
     
     
         15 . The recombinant virus of  claim 13 , wherein the therapeutic agent is an anti-VEGF single chain antibody, a plasminogen K5 domain, a human tissue factor-αvβ3-integrin RGD fusion protein, interleukin-24 or an IL-6-IL-6 receptor fusion protein. 
     
     
         16 . A combination, comprising:
 a recombinant virus of  claim 1 ; and   a substrate transported into a cell that expresses the transporter; and/or an anti-cancer compound.   
     
     
         17 . The combination of  claim 16 , wherein the substrate is detectable. 
     
     
         18 . The combination of  claim 16 , wherein the substrate emits an electromagnetic signal or induces such emission. 
     
     
         19 . The combination of  claim 16 , wherein the substrate is radiolabeled. 
     
     
         20 . The combination of  claim 16 , wherein the substrate is conjugated to a cytotoxic agent. 
     
     
         21 . The combination of  claim 20 , wherein conjugation is direct or via a linker. 
     
     
         22 . The combination of  claim 20 , wherein the cytotoxic agent is a radiolabel, a cytotoxin or a cytotoxic drug. 
     
     
         23 . The combination of  claim 20 , wherein the cytotoxic agent is selected from among double-chain ricin, ricin A chain, abrin, abrin A chain, saporin, modeccin, modeccin A chain,  Pseudomonas aeruginosa  exotoxin, Cholera toxin, Shigella toxin,  E. coli  heat labile toxin, Diptheria toxin, doxorubicin, daunomycin, fluorouracil, methotrexate, taxol, ricin A, colchicine, cytochalasins, monensin, ouabain, mitoxanthrone, vindesine, vinblastine, vincristine or enterotoxin. 
     
     
         24 . The combination of  claim 16 , wherein the substrate is conjugated to an anticancer agent. 
     
     
         25 . The combination of  claim 16 , wherein the combination comprises the virus and an anti-cancer compound selected from among a cytokine, a chemokine, a growth factor, a photosensitizing agent, a toxin, an anti-cancer antibiotic, a chemotherapeutic compound, a radionuclide, an angiogenesis inhibitor, a signaling modulator, an anti-metabolite, an anti-cancer vaccine, an anti-cancer oligopeptide, a mitosis inhibitor protein, an antimitotic oligopeptide, an anti-cancer antibody, an anti-cancer antibiotic, an immunotherapeutic agent, a bacterium and a combination of any of the preceding thereof. 
     
     
         26 . The combination of  claim 25 , wherein the anti-cancer compound is selected from among cisplatin, carboplatin, gemcitabine, irinotecan, an anti-EGFR antibody and an anti-VEGF antibody. 
     
     
         27 . The combination of  claim 16 , wherein the substrate and virus are formulated as a single composition or separately in two compositions. 
     
     
         28 . The combination of  claim 16 , wherein the anti-cancer compound and virus are formulated as a single composition or separately in two compositions. 
     
     
         29 . A kit, comprising the combination of  claim 16 ; and optionally instructions for administration of the composition(s). 
     
     
         30 . A pharmaceutical composition, comprising a recombinant virus of  claim 1  in a pharmaceutically acceptable carrier. 
     
     
         31 . The pharmaceutical composition of  claim 30  that is formulated for local or systemic administration. 
     
     
         32 . A vaccine, comprising the recombinant virus of  claim 1 . 
     
     
         33 . The vaccine of  claim 32  that is a smallpox vaccine. 
     
     
         34 . A method of imaging or detecting a tumor, an inflammation or a wound within a subject comprising:
 administering a vaccinia virus of  claim 1  to a subject having a tumor;   administering a detectable substrate that is transported into a cell that expresses the transporter encoded by the virus; and   detecting the accumulation of the substrate, whereby the tumor, wound and/or inflammation is imaged or detected.   
     
     
         35 . The method of  claim 34 , wherein the substrate is detected by fluorescence imaging, magnetic resonance imaging (MRI), single-photon emission computed tomography (SPECT), positron emission tomography (PET), scintigraphy, gamma camera, a β+ detector, a γ detector or a combination thereof. 
     
     
         36 . A method of treatment, comprising administering a vaccinia virus of  claim 1  to a subject to effect treatment. 
     
     
         37 . The method of  claim 36 , wherein the subject is treated for a tumor, cancer or metastasis. 
     
     
         38 . The method of  claim 36 , further comprising:
 administering a substrate that is transported into a cell that expresses the transporter encoded by the virus, wherein:   the substrate is therapeutic or is conjugated to a therapeutic agent, whereby treatment is effected; and   the substrate is administered before or after or simultaneously with the virus.   
     
     
         39 . The method of  claim 38 , wherein the subject has a tumor and the substrate or substrate conjugate is effective in treating a tumor. 
     
     
         40 . The method of  claim 38 , wherein the substrate is conjugated to a cytotoxic agent. 
     
     
         41 . The method of  claim 40 , wherein the cytotoxic agent is a radiolabel, a cytotoxin or a cytotoxic drug. 
     
     
         42 . The method of  claim 41 , wherein the cytotoxic agent is selected from among double-chain ricin, ricin A chain, abrin, abrin A chain, saporin, modeccin, modeccin A chain,  Pseudomonas aeruginosa  exotoxin, Cholera toxin, Shigella toxin,  E. coli  heat labile toxin and Diptheria toxin, doxorubicin, daunomycin, 5-fluorouracil, methotrexate, taxol, ricin A, colchicine, cytochalasins, monensin, ouabain, mitoxanthrone, vindesine, vinblastine, vincristine or enterotoxin. 
     
     
         43 . The method of  claim 38 , wherein the substrate is conjugated to an anticancer agent. 
     
     
         44 . The method of  claim 36 , wherein the virus is administered systemically, intravenously, intraarterially, intratumorally, endoscopically, intralesionally, intramuscularly, intradermally, intraperitoneally, intravesicularly, intraarticularly, intrapleurally, percutaneously, subcutaneously, orally, parenterally, intranasally, intratracheally, by inhalation, intracranially, intraprostaticaly, intravitreally, topically, ocularly, vaginally, or rectally. 
     
     
         45 . The method of  claim 36 , further comprising administering an anticancer agent or treatment, wherein the anti-cancer agent or treatment is administered before or after or simultaneously with the virus or the virus and substrate. 
     
     
         46 . The method of  claim 45 , wherein:
 an anticancer agent is administered; and   the anticancer agent is selected from among a cytokine, a chemokine, a growth factor, a photosensitizing agent, a toxin, an anti-cancer antibiotic, a chemotherapeutic compound, a radionuclide, an angiogenesis inhibitor, a signaling modulator, an anti-metabolite, an anti-cancer vaccine, an anti-cancer oligopeptide, a mitosis inhibitor protein, an antimitotic oligopeptide, an anti-cancer antibody, an anti-cancer antibiotic, an immunotherapeutic agent, hyperthermia or hyperthermia therapy, a bacterium, radiation therapy and a combination of any of the preceding thereof.   
     
     
         47 . The method of  claim 46 , wherein the anticancer agent is selected from among cisplatin, carboplatin, gemcitabine, irinotecan, an anti-EGFR antibody and an anti-VEGF antibody. 
     
     
         48 . The method of  claim 45 , wherein the anticancer agent is administered simultaneously or intermittently with the virus or with the virus and/or substrate. 
     
     
         49 . The method of  claim 45 , wherein the virus and the anticancer agent are administered as a single composition or as two compositions, or the virus, substrate and anticancer agent are administered in a single composition or in two compositions containing two of the substrate, virus and anticancer agent, or in three compositions. 
     
     
         50 . The method of  claim 37 , wherein the subject is treated for a human tumor selected from among a bladder tumor, breast tumor, prostate tumor, carcinoma, basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain cancer, CNS cancer, glioma tumor, cervical cancer, choriocarcinoma, colon and rectum cancer, connective tissue cancer, cancer of the digestive system, endometrial cancer, esophageal cancer, eye cancer, cancer of the head and neck, gastric cancer, intra-epithelial neoplasm, kidney cancer, larynx cancer, leukemia, liver cancer, lung cancer, lymphoma, Hodgkin's lymphoma, Non-Hodgkin's lymphoma, melanoma, myeloma, neuroblastoma, oral cavity cancer, ovarian cancer, pancreatic cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer, renal cancer, cancer of the respiratory system, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, uterine cancer, and cancer of the urinary system. 
     
     
         51 . The method of  claim 37 , wherein the subject is treated for a tumor and the tumor is an ovarian tumor, a breast tumor, a pancreatic tumor, a colon tumor, or a lung tumor. 
     
     
         52 . The method of  claim 37 , wherein the subject is treated for a tumor and the tumor is selected from among lymphosarcoma, osteosarcoma, mammary tumors, mastocytoma, brain tumor, melanoma, adenosquamous carcinoma, carcinoid lung tumor, bronchial gland tumor, bronchiolar adenocarcinoma, fibroma, myxochondroma, pulmonary sarcoma, neurosarcoma, osteoma, papilloma, retinoblastoma, Ewing's sarcoma, Wilm's tumor, Burkitt's lymphoma, microglioma, neuroblastoma, osteoclastoma, oral neoplasia, fibrosarcoma, osteosarcoma and rhabdomyosarcoma, genital squamous cell carcinoma, transmissible venereal tumor, testicular tumor, seminoma, Sertoli cell tumor, hemangiopericytoma, histiocytoma, chloroma, granulocytic sarcoma, corneal papilloma, corneal squamous cell carcinoma, hemangiosarcoma, pleural mesothelioma, basal cell tumor, thymoma, stomach tumor, adrenal gland carcinoma, oral papillomatosis, hemangioendothelioma, cystadenoma, follicular lymphoma, intestinal lymphosarcoma, fibrosarcoma, and pulmonary squamous cell carcinoma, leukemia, hemangiopericytoma, ocular neoplasia, preputial fibrosarcoma, ulcerative squamous cell carcinoma, preputial carcinoma, connective tissue neoplasia, mastocytoma, hepatocellular carcinoma, lymphoma, pulmonary adenomatosis, pulmonary sarcoma, Rous sarcoma, reticulo-endotheliosis, fibrosarcoma, nephroblastoma, B-cell lymphoma, lymphoid leukosis, retinoblastoma, hepatic neoplasia, lymphosarcoma, plasmacytoid leukemia, swimbladder sarcoma (in fish), caseous lumphadenitis, lung carcinoma, insulinoma, lymphoma, sarcoma, neuroma, pancreatic islet cell tumor, gastric MALT lymphoma and gastric adenocarcinoma. 
     
     
         53 . The method of  claim 36 , further comprising administering an anti-viral agent. 
     
     
         54 . The method of  claim 53 , wherein the antiviral agent is selected from among cidofovir, alkoxyalkyl esters of cidofovir, imatinib, gancyclovir, acyclovir and Tecovirimat. 
     
     
         55 . The method of  claim 36 , further comprising:
 administering a detectable substrate that is transported into a cell that expresses the transporter encoded by the virus; and   detecting the substrate or accumulation of the substrate, whereby the therapeutic treatment of the tumor is monitored.   
     
     
         56 . The method of  claim 38 , further comprising:
 detecting the substrate or accumulation of the substrate, whereby the therapeutic treatment of the tumor is monitored.   
     
     
         57 . A method of detection of a tumor or metastasis and treatment a tumor and/or metastases within a subject, comprising:
 administering a vaccinia virus of  claim 1  to a subject having a tumor, wherein the virus optionally encodes an anti-tumor therapeutic agent;   optionally administering a therapeutic agent to the subject for treatment of a tumor;   administering a detectable substrate that is transported into a cell that expresses the transporter encoded by the virus; and   detecting substrate or the accumulation of the substrate to detect a tumor or metastases and to effect treatment thereof.   
     
     
         58 . The method of  claim 57 , wherein detection is monitored as a function of time, whereby treatment is monitored. 
     
     
         59 . An isolated cell, comprising a recombinant virus of  claim 1 .