Humanized Viral Vectors and Methods of Use Thereof

Abstract

The present invention provides humanized viral vectors and methods of use thereof for delivery of transgenes or therapeutic nucleic acids to human subjects. Humanized viral vectors are modified from known viral vectors such as those based on AAV by coating their surface with a human protein such as human serum albumin and optionally a lipid coating or formulation, so that the foreign or non-human nature of the vector is masked. The coating is performed in a manner that reduces or prevents binding of antibodies to the vector surface, thereby reducing or preventing antibody-mediated clearance of vector, but still allowing the vector to transduce target cells and achieve therapeutic gene transfer. Such humanized vectors therefore evade pre-existing immune surveillance, reduce immune responses, and achieve therapeutic gene transfer in the presence of pre-existing antibodies to the viral vector.

1 . A humanized viral vector encoding a therapeutic molecule of interest, said vector comprising at least one human surface molecule thereby reducing antibody binding and clearance via immune surveillance. 
   
   
       2 . The viral vector of  claim 1 , wherein said therapeutic molecule of interest is a nucleic acid encoding a protein or peptide which upon expression, confers a beneficial therapeutic effect. 
   
   
       3 . The viral vector of  claim 1 , wherein said therapeutic molecule of interest is a therapeutic nucleic acid which upon expression modulates expression of a cognate mRNA. 
   
   
       4 . The viral vector of  claim 1 , which is an adeno-associated viral vector comprising capsid proteins. 
   
   
       5 . The viral vector of  claim 4 , comprising human proteins affixed to said capsid protein. 
   
   
       6 . The viral vector of  claim 4  or  5 , encapsulated in a lipid formulation optionally comprising carbohydrates. 
   
   
       7 . The viral vector of  claim 6 , wherein said human protein is human serum albumin. 
   
   
       8 . The viral vector of  claim 6  wherein the human protein is a complex of a protein and a ligand. 
   
   
       9 . The viral vector of  claim 8 , wherein said complex comprises human serum albumin and heparin. 
   
   
       10 . The viral vector of  claim 8 , wherein said complex comprises human serum albumin and a fatty acid. 
   
   
       11 . The viral vector of  claim 7 , wherein said complex is an encapsulating lipid formulation optionally comprising a carbohydrate moiety which enhances target cell uptake of said vector. 
   
   
       12 . The vector of  claim 11 , wherein said lipid comprises 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine. 
   
   
       13 . The vector of  claim 11  wherein said complex is phosphoethanolamine-N-lactosyl. 
   
   
       14 . The viral vector of  claim 2 , wherein said protein or peptide is selected from the group consisting of a cystic fibrosis transmembrane regulator protein, dystrophin, utrophin, Factor XIII, Factor IX, Factor X, a monoclonal antibody, erythropoietin, the LDL receptor, lipoprotein lipase, ornithine transcarbamylase, β-globin, α-globin, spectrin, α-antitrypsin, a human apolipoprotein, adenosine deaminase, hypoxanthine guanine phosphoribosyl transferase, β-glucocerebrosidase, sphingomyelinase, lysosomal hexosaminidase, branched-chain keto acid dehydrogenase, a hormone, insulin-like growth factors 1 and 2, platelet derived growth factor, epidermal growth factor, nerve growth factor, neurotrophic factor-3 and -4, brain-derived neurotrophic factor, glial derived growth factor, transforming growth factor α and β, a cytokine, α-interferon, β-interferon, interferon-γ, interleukin-2, interleukin-4, interleukin 12, granulocyte-macrophage colony stimulating factor, lymphotoxin, a suicide gene product, herpes simplex virus thymidine kinase, cytosine deaminase, diphtheria toxin, cytochrome P450, deoxycytidine kinase, tumor necrosis factor, a tumor suppressor gene product, p53, Rb, Wt-1, NF1, VHL, and APC. 
   
   
       15 . The viral vector of  claim 6 , wherein said protein or peptide is selected from the group consisting of cystic fibrosis transmembrane regulator protein, dystrophin, utrophin, Factor XIII, Factor IX, Factor X, a monoclonal antibody, erythropoietin, the LDL receptor, lipoprotein lipase, ornithine transcarbamylase, β-globin, α-globin, spectrin, a human apolipoprotein, α-antitrypsin, adenosine deaminase, hypoxanthine guanine phosphoribosyl transferase, β-glucocerebrosidase, sphingomyelinase, lysosomal hexosaminidase, branched-chain keto acid dehydrogenase, a hormone, insulin-like growth factors 1 and 2, platelet derived growth factor, epidermal growth factor, nerve growth factor, neurotrophic factor-3 and -4, brain-derived neurotrophic factor, glial derived growth factor, transforming growth factor α and β, a cytokine, α-interferon, β-interferon, interferon-γ, interleukin-2, interleukin-4, interleukin 12, granulocyte-macrophage colony stimulating factor, lymphotoxin, a suicide gene product, herpes simplex virus thymidine kinase, cytosine deaminase, diphtheria toxin, cytochrome P450, deoxycytidine kinase, tumor necrosis factor, a tumor suppressor gene product, p53, Rb, Wt-1, NF1, VHL, and APC. 
   
   
       16 . The viral vector of  claim 12 , wherein said protein or peptide is selected from the group consisting of cystic fibrosis transmembrane regulator protein, dystrophin, utrophin, Factor XIII, Factor IX, Factor X, a monoclonal antibody, erythropoietin, the LDL receptor, lipoprotein lipase, ornithine transcarbamylase, β-globin, α-globin, spectrin, a human apolipoprotein, α-antitrypsin, adenosine deaminase, hypoxanthine guanine phosphoribosyl transferase, β-glucocerebrosidase, sphingomyelinase, lysosomal hexosaminidase, branched-chain keto acid dehydrogenase, a hormone, insulin-like growth factors 1 and 2, platelet derived growth factor, epidermal growth factor, nerve growth factor, neurotrophic factor-3 and -4, brain-derived neurotrophic factor, glial derived growth factor, transforming growth factor α and β, a cytokine, α-interferon, β-interferon, interferon-γ, interleukin-2, interleukin-4, interleukin 12, granulocyte-macrophage colony stimulating factor, lymphotoxin, a suicide gene product, herpes simplex virus thymidine kinase, cytosine deaminase, diphtheria toxin, cytochrome P450, deoxycytidine kinase, tumor necrosis factor, a tumor suppressor gene product, p53, Rb, Wt-1, NF1, VHL, and APC. 
   
   
       17 . A pharmaceutical composition comprising the viral vector of  claim 13 ,  14 ,  15 , or  16  in a pharmaceutically acceptable carrier. 
   
   
       18 . A method for delivering a transgene to a cell comprising administration of an effective amount of the pharmaceutical composition of  claim 17  to a patient in need thereof. 
   
   
       19 . The method of  claim 18 , wherein said patient has a clotting disorder and said transgene encodes a clotting factor. 
   
   
       20 . The method of  claim 19 , wherein said clotting factor is hFIX. 
   
   
       21 . A pharmaceutical preparation comprising a plurality of viral vectors encapsulated in a lipid formulation studded with human surface proteins and optionally at least one carbohydrate moiety.