Oncolytic adenovirus armed with therapeutic genes

Abstract

The present invention involves compositions and methods for treating cancer using a mutant adenovirus comprising a polynucleotide encoding a therapeutic polypeptide that is targeted to cells with a mutant retinoblastoma pathway. The mutant adenovirus is able to kill the tumor cells without harming cells with a wild type retinoblastoma pathway.

1 . A method for treating a brain tumor in a patient comprising: 
 a) identifying a patient having a brain tumor; and    b) contacting the tumor with an oncolytic adenovirus that encodes a therapeutic gene and an E1A polypeptide that cannot bind Rb.    
     
     
         2 . The method of  claim 1 , wherein the oncolytic adenovirus is a Delta 24 adenovirus.  
     
     
         3 . The method of  claim 1 , wherein the oncolytic adenovirus comprises a targeting moiety.  
     
     
         4 . The method of  claim 3 , wherein the targeting moiety is a chimeric adenoviral fiber protein.  
     
     
         5 . The method of  claim 4 , wherein the chimeric adenoviral fiber protein comprises an RGD amino acid sequence.  
     
     
         6 . The method of  claim 4 , wherein the chimeric adenoviral fiber protein comprises a vIII amino acid sequence.  
     
     
         7 . The method of  claim 4 , wherein the chimeric adenoviral fiber protein comprises a PEPHC1 amino acid sequence.  
     
     
         8 . The method of  claim 1 , wherein the therapeutic gene is Rb, CFTR, p16, p21, p27, p57, p73, C-CAM, APC, CTS-1, zac1, scFV ras, DCC, NF-1, NF-2, WT-1, MEN-I, MEN-II, BRCA1, VHL, MMAC1, FCC, MCC, BRCA2, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11 IL-12, GM-CSF, G-CSF, thymidine kinase, mda7, fus, interferon α, interferon β, interferon γ, ADP, p53, ABL1, BLC1, BLC6, CBFA1, CBL, CSFIR, ERBA, ERBB, EBRB2, ETS1, ETS2, ETV6, FGR, FOX, FYN, HCR, HRAS, JUN, KRAS, LCK, LYN, MDM2, MLL, MYB, MYC, MYCL1, MYCN, NRAS, PIM1, PML, RET, SRC, TAL1, TCL3, YES, MADH4, RB1, TP53, WT1, TNF, BDNF, CNTF, NGF, IGF, GMF, aFGF, bFGF, NT3, NT5, ApoAI, ApoAIV, ApoE, Rap1A, cytosine deaminase, Fab, ScFv, BRCA2, zac1, ATM, HIC-1, DPC-4, FHIT, PTEN, ING1, NOEY1, NOEY2, OVCA1, MADR2, 53BP2, IRF-1, Rb, zac1, DBCCR-1, rks-3, COX-1, TFPI, PGS, Dp, E2F, ras, myc, neu, raf, erb, fms, trk, ret, gsp, hst, abl, E1A, p300, VEGF, FGF, thrombospondin, BAI-1, GDAIF, or MCC.  
     
     
         9 . The method of  claim 1 , wherein the therapeutic gene is yeast cytosine deaminase.  
     
     
         10 . The method of  claim 1 , wherein the therapeutic gene is Ang-2.  
     
     
         11 . The method of  claim 1 , wherein the therapeutic gene is NIS.  
     
     
         12 . The method of  claim 1 , wherein the tumor is contacted with the adenovirus by delivery of the adenovirus intracranially into the patient.  
     
     
         13 . The method of  claim 1 , wherein the tumor is a astrocytoma, oligodendroglioma, anaplastic glioma, glioblastoma, ependymoma, meningioma, pineal region tumor, choroid plexus tumor, neuroepithelial tumor, embryonal tumor, peripheral neuroblastic tumor, tumor of cranial nerves, tumor of the hemopoietic system, germ cell tumors, or tumor of the sellar region.  
     
     
         14 . The method of  claim 13 , wherein the tumor is a glioblastoma.  
     
     
         15 . A method of treating cancer in a patient comprising: 
 a) administering to a patient an effective amount of a composition comprising an oncolytic adenovirus comprising (i) a nucleic acid, wherein the nucleotides encoding amino acids 122-129 of the encoded E1A polypeptide are deleted, and (ii) an expression cassette comprising a polynucleotide encoding a yeast cytosine deaminase; and    b) administering an effective amount of a pro-drug, wherein the pro-drug is metabolized to a cytotoxic drug by a polypeptide encoded by the yeast cytosine deaminase.    
     
     
         16 . The method of  claim 15 , wherein the yeast cytosine deaminase is a humanized yeast cytosine deaminase.  
     
     
         17 . The method of  claim 15 , wherein the oncolytic adenovirus comprises a targeting moiety.  
     
     
         18 . The method of  claim 17 , wherein the targeting moiety is a chimeric adenoviral fiber protein.  
     
     
         19 . The method of  claim 17 , wherein the chimeric adenoviral fiber protein comprises an RGD amino acid sequence.  
     
     
         20 . The method of  claim 18 , wherein the chimeric adenoviral fiber protein comprises a vIII amino acid sequence.  
     
     
         21 . The method of  claim 18 , wherein the chimeric adenoviral fiber protein comprises a PEPHC1 amino acid sequence.  
     
     
         22 . The method of  claim 16 , wherein the cell is a tumor cell.  
     
     
         23 . The method of  claim 16 , wherein the cancer is a astrocytoma, oligodendroglioma, anaplastic glioma, glioblastoma, ependymoma, meningioma, pineal region tumor, choroid plexus tumor, neuroepithelial tumor, embryonal tumor, peripheral neuroblastic tumor, tumor of cranial nerves, tumor of the hemopoietic system, germ cell tumors, or tumor of the sellar region.  
     
     
         24 . The method of  claim 23 , wherein the tumor is a glioblastoma.  
     
     
         25 . The method of  claim 15 , wherein the oncolytic adenovirus is suitably dispersed in a pharmacologically acceptable formulation.  
     
     
         26 . The method of  claim 15 , wherein the composition is administered intracranially.  
     
     
         27 . The method of  claim 26 , wherein the composition is directly injected into a tumor.  
     
     
         28 . The method of  claim 26 , wherein the administration occurs more than once.  
     
     
         29 . The method of  claim 28 , wherein the composition is administered at least three times to the patient.  
     
     
         30 . The method of  claim 15 , further comprising administering to the patient a second therapy, wherein the second therapy is anti-angiogenic therapy, chemotherapy, immunotherapy, surgery, radiotherapy, immunosuppresive agents, or gene therapy with a therapeutic polynucleotide.  
     
     
         31 . The method of  claim 30 , wherein the second therapy is administered to the patient before administration of the composition comprising the oncolytic adenovirus.  
     
     
         32 . The method of  claim 30 , wherein the second therapy is administered to the patient at the same time as administration of the composition comprising the oncolytic adenovirus.  
     
     
         33 . The method of  claim 30 , wherein the second therapy is administered to the patient after administration of the composition comprising the oncolytic adenovirus.  
     
     
         34 . The method of  claim 30 , wherein the chemotherapy comprises an alkylating agent, mitotic inhibitor, antibiotic, or antimetabolite.  
     
     
         35 . The method of  claim 30 , wherein the chemotherapy comprises CPT-11, temozolomide, or a platin compound.  
     
     
         36 . The method of  claim 30 , wherein radiotherapy comprises X-ray irradiation, UV-irradiation, γ-irradiation, or microwaves.  
     
     
         37 . The method of  claim 15 , wherein from about 10 3  to about 10 15  viral particles are administered to the patient.  
     
     
         38 . The method of  claim 37 , wherein from about 10 5  to about 10 12  viral particles are administered to the patient.  
     
     
         39 . The method of  claim 37 , wherein from about 10 7  to about 10 10  viral particles are administered to the patient.