Hybrid adeno-retroviral vector for the transfection of cells

Abstract

An adenovirus, including adenoviral capsid proteins, and a replication-defective adenoviral vector that includes a 5′ retroviral LTR nucleic acid sequence, a 3′ retroviral LTR nucleic acid sequence, a nucleic acid sequence encoding a portion of a retroviral envelope protein adjacent to either the 5′ LTR or the 3′ LTR nucleic acid sequence, a retroviral packaging sequence and a nucleic acid sequence encoding a transgene located between the 5′ LTR and the 3′ LTR is provided. Host cells infected with this adenovirus are also provided. An adenoviral vector is provided that includes an adenoviral polynucleotide sequence comprising a nucleic acid encoding a transgene, a retroviral packaging signal, a 5′ and a 3′ retroviral LTR, and a portion of a retroviral envelope polypeptide, wherein the adenoviral polynucleotide sequence does not encode one or more of E1, E3 or E4. A method for transforming a cell is also provided using a virus or a vector of the invention, as is a method for introducing a transgene into a cell that is not able to produce viral particles with a single viral vector. A method is also provided for preventing or treating disorder in a subject using the adenoviral vectors of the invention. A pharmaceutical composition is also provided that includes an adenoviral vector of the invention and a pharmaceutically acceptable carrier.

1 . A pharmaceutical composition, comprising:
 (a) an adenoviral vector, wherein the adenoviral vector comprises a 5′ retroviral LTR nucleic acid sequence, wherein the 5′ retroviral LTR nucleic acid sequence comprises a U3, R, U5 nucleic acid sequence and a portion of a nucleic acid sequence encoding a retroviral envelope polypeptide,   a 3′ retroviral LTR nucleic acid sequence, wherein the 3′ retroviral LTR comprises an U3, R, U5 nucleic acid sequence and a portion of a nucleic acid sequence encoding a retroviral envelope polypeptide, wherein the 5′ and 3′ retroviral LTR nucleic acid sequences are Moloney Leukemia Virus LTR nucleic acid sequences,   a nucleic acid sequence encoding a transgene located between the 5′ LTR and the 3′ LTR, and   a retroviral packaging sequence,   wherein the 5′ retroviral LTR, the 3′ retroviral LTR, the transgene and the retroviral packaging sequence are inserted into a replication-defective adenoviral vector nucleic acid sequence, and wherein the vector does not comprise a nucleic acid sequence encoding a functional retroviral gag or a functional retroviral pol, or both; and   (b) a pharmaceutically acceptable carrier.   
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the transgene encodes erythropoietin. 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein the adenoviral vector nucleic acid comprises a functional deletion of a nucleic acid sequence encoding an early transcriptional unit polypeptide. 
     
     
         4 . The adenoviral vector of  claim 3 , wherein the functional deletion is a mutation or a deletion of a nucleic acid sequence encoding an early transcriptional unit polypeptide. 
     
     
         5 . The adenoviral vector of  claim 4 , wherein the nucleic acid sequence encoding an early transcriptional unit polypeptide is E1. 
     
     
         6 . The adenovirus of  claim 5 , further comprising a deletion in a nucleic acid sequence encoding an E3 early transcriptional unit polypeptide. 
     
     
         7 . The adenoviral vector of  claim 4 , wherein the 5′ retroviral LTR nucleic acid sequence, the 3′ retroviral LTR nucleic acid sequence, nucleic acid sequence encoding the transgene, the nucleic acids encoding portions of the retroviral envelope polypeptide, and the retroviral packaging sequence are inserted into the nucleic acid sequence encoding the early transcriptional unit polypeptide. 
     
     
         8 . The adenoviral vector of  claim 2 , further comprising a promoter operably linked to the transgene. 
     
     
         9 . The adenoviral vector of  claim 8 , wherein the promoter is a cytomegalovirus (CMV) promoter.