Detargeted adenovirus variants and related methods

Abstract

The present disclosure describes the generation and the use of Ad variants (Ad) possessing any combination of mutations in genes that code for the hexon, penton, fiber, and non-structural proteins, where simultaneous modification of hexon and penton are made to avoid the trapping of Ad in the liver and to reduce toxicity after intravascular virus administration. Such liver de-targeted Ad can be useful tool for selective and specific gene delivery to extra-hepatic tissues and cells, including disseminated metastatic cancer cells.

1 .- 40 . (canceled) 
     
     
         41 . An isolated adenovirus, comprising:
 a mutation in the RGD motif of the penton protein, wherein the mutation causes reduced binding of β3 integrins of a host cell; and   two mutations in the hexon hypervariable loops, wherein said mutations in the hexon hypervariable loops comprise a mutation in the HVR1 loop and a mutation in the HVR7 loop,   wherein the mutation in the HVR1 loop of the hexon protein causes reduced virus trapping in Kupffer cells,   wherein the combination of the mutation in the RGD motif, the mutation in the HVR1 loop and the mutation in the HVR7 loop causes reduced liver sequestration of the adenovirus, and   wherein the adenovirus is a replication-deficient adenovirus.   
     
     
         42 . The adenovirus of  claim 41 , wherein the adenovirus comprises a mutation or deletion in E1, E2, E3, E4, and/or L1-L5 regions of adenovirus genome. 
     
     
         43 . The adenovirus of  claim 41 , wherein the adenovirus comprises a mutation or deletion in E1 region of adenovirus genome. 
     
     
         44 . The adenovirus of  claim 41 , wherein the adenovirus comprises a mutation or deletion in E1 and E4 regions of adenovirus genome. 
     
     
         45 . The adenovirus of  claim 41 , wherein the adenovirus comprises a mutation or deletion in E1, E2 and E3 regions of adenovirus genome. 
     
     
         46 . The adenovirus of  claim 41 , wherein the adenovirus comprises a mutation or deletion in E1, E3 and E4 regions of adenovirus genome. 
     
     
         47 . The adenovirus of  claim 41 , wherein the adenovirus comprises a mutation or deletion in E1, E2, E3 and E4 regions of adenovirus genome. 
     
     
         48 . The adenovirus of  claim 41 , wherein the adenovirus comprises a mutation or deletion in E1, E2, E3, E4 and L1-L5 regions of adenovirus genome. 
     
     
         49 . The adenovirus of  claim 41 , wherein the adenovirus comprises a complete deletion of viral coding sequences of adenovirus genome. 
     
     
         50 . The adenovirus of  claim 41 , wherein the RDG motif of the penton loop is substituted with a non-RGD motif-containing peptide capable of binding non-β3 cellular integrins. 
     
     
         51 . The adenovirus of  claim 41 , wherein the adenovirus is a species B, C, or D adenovirus. 
     
     
         52 . The adenovirus of  claim 41 , wherein the adenovirus further comprises a mutation in the HVR3 or HVR5 region of the hexon protein. 
     
     
         53 . The adenovirus of  claim 52 , wherein the mutation in the HVR3 or HVR5 region of the hexon protein is a substitution. 
     
     
         54 . The adenovirus of  claim 41 , wherein the single virus genomic DNA molecule comprises a nucleotide sequence that encodes SEQ ID NO:2. 
     
     
         55 . The adenovirus of  claim 41 , wherein the mutation in the HVR1 region of the hexon protein is a substitution. 
     
     
         56 . The adenovirus of  claim 41 , wherein the adenovirus comprises a gene of interest selected from a cytokine, a cellular receptor, a nuclear receptor, a ligand, a coagulation factor, a CFTR protein, insulin, dystrophin, a growth hormone, immune-stimulatory or immune-suppressing protein of eukaryotic or prokaryotic origin, of an enzyme, an enzyme inhibitor, a polypeptide with antitumor effect, a polypeptide capable of inhibiting a bacterial, parasitic or viral infection, an antibody, a toxin, an immunotoxin, shRNA, LncRNA, ribozyme, or a marker. 
     
     
         57 . A pharmaceutical composition comprising:
 the adenovirus according to  claim 41 ; and   a pharmaceutically acceptable carrier.   
     
     
         58 . A method of delivering a gene to a non-hepatic mammalian cell in a subject, comprising the step of administering to the subject an effective amount of the adenovirus of  claim 41 . 
     
     
         59 . The method of  claim 58 , wherein the adenovirus is administered intravenously. 
     
     
         60 . The method of  claim 58 , wherein the adenovirus comprises a complete deletion of viral coding sequences of adenovirus genome.