Gene delivery vectors provided with a tissue tropism for smooth muscle cells, and/or endothelial cells

Abstract

A gene delivery vehicle having been provided with at least a tissue tropism for cells selected from the group of smooth muscle cells, endothelial cells, and/or liver cells. The tissue tropism is generally provided by a virus capsid, such as one comprising protein fragments from at least two different viruses, such as two different adenoviruses, including adenovirus of subgroup C or subgroup B (for example, adenovirus 16). The protein fragments can comprise a tissue tropism-determining fragment of a fiber protein derived from a subgroup B adenovirus. Also, cells for producing such gene delivery vehicles and pharmaceutical compositions containing these gene delivery vehicles are provided. Further, a method is disclosed for delivering nucleic acid to cells such as smooth muscle cells and/or endothelial cells which involves administering to the cells an adenovirus capsid having proteins from at least two different adenoviruses and wherein at least a tissue tropism-determining fragment of a fiber protein is derived from a subgroup B adenovirus. Particular constructs are also disclosed.

1 . A viral gene delivery vehicle comprising at least a tissue tropism for cells selected from the group consisting of smooth muscle cells, endothelial cells, or smooth muscle cells and epothelial cells.  
     
     
         2 . The viral gene delivery vehicle of  claim 1 , wherein said gene delivery vehicle has been deprived of at least a tissue tropism for liver cells.  
     
     
         3 . The viral gene delivery vehicle of  claim 1 , wherein said tissue tropism is provided by a virus capsid.  
     
     
         4 . The viral gene delivery vehicle of  claim 3 , wherein said virus capsid comprises protein fragments from at least two different viruses.  
     
     
         5 . The viral gene delivery vehicle of  claim 4 , wherein at least one of said viruses is an adenovirus.  
     
     
         6 . The viral gene delivery vehicle of  claim 4 , wherein at least one of said viruses is an adenovirus of subgroup B.  
     
     
         7 . The viral gene delivery vehicle of  claim 4 , wherein at least one of said protein fragments comprises a tissue tropism-determining fragment of a fiber protein derived from a subgroup B adenovirus.  
     
     
         8 . The viral gene delivery vehicle of  claim 6 , wherein said subgroup B adenovirus is adenovirus 16.  
     
     
         9 . The viral gene delivery vehicle of  claim 6 , wherein protein fragments not derived from an adenovirus of subgroup B are derived from an adenovirus of subgroup C.  
     
     
         10 . The viral gene delivery vehicle of  claim 1 , further comprising a nucleic acid derived from an adenovirus.  
     
     
         11 . The viral gene delivery vehicle of  claim 1 , further comprising a nucleic acid derived from at least two different adenoviruses.  
     
     
         12 . The viral gene delivery vehicle of  claim 10 , wherein said nucleic acid comprises at least one sequence encoding a fiber protein comprising at least a tissue tropism-determining fragment of a subgroup B adenovirus fiber protein.  
     
     
         13 . The viral gene delivery vehicle of  claim 10 , wherein said nucleic acid derived from adenovirus is modified such that the capacity of said adenovirus nucleic acid to replicate in a target cell has been reduced or disabled.  
     
     
         14 . The viral gene delivery vehicle of  claim 10 , wherein said nucleic acid derived from an adenovirus is modified such that a host immune system's capacity to mount an immune response against adenoviral proteins encoded by adenoviral nucleic acid has been reduced or disabled.  
     
     
         15 . The viral gene delivery vehicle of  claim 1 , further comprising a minimal adenovirus vector or an Ad/AAV chimeric vector.  
     
     
         16 . The viral gene delivery vehicle of  claim 1 , further comprising at least one non-adenoviral nucleic acid.  
     
     
         17 . The viral gene delivery vehicle of  claim 16 , wherein at least one of said non-adenoviral nucleic acids is a gene selected from the group of genes encoding a protein selected from the group consisting of: an apolipoprotein, a nitric oxide synthase, a herpes simplex virus thymidine kinase, an interleukin-3, an interleukin-1α, an (anti) angiogenesis protein, an anti-proliferation protein, a smooth muscle cell anti-migration protein, a vascular endothelial growth factor (VGEF), a basic fibroblast growth factor, a hypoxia inducible factor 1α (HIF-1α) and a PAI-1.  
     
     
         18 . The viral gene delivery vehicle of  claim 5 , wherein at least one of said viruses is an adenovirus of subgroup B.  
     
     
         19 . The viral gene delivery vehicle of  claim 7 , wherein said subgroup B adenovirus is adenovirus 16.  
     
     
         20 . A recombinant adenovirus of a subgroup C origin having a reduced tissue tropism for liver cells as compared to the corresponding wild type adenovirus of subgroup C origin, said recombinant adenovirus comprising: 
 a capsid comprising a chimeric fiber protein, wherein a knob domain of said chimeric fiber protein is of an adenovirus origin selected from the group consisting of adenovirus 12, adenovirus 16, adenovirus 28 and adenovirus 40-L.    
     
     
         21 . A pharmaceutical composition comprising: 
 the recombinant adenovirus of  claim 20;  and    a suitable vehicle.    
     
     
         22 . The recombinant adenovirus of  claim 20 , wherein said reduced tissue tropism is provided by a virus capsid.  
     
     
         23 . The recombinant adenovirus of  claim 22 , wherein said virus capsid comprises protein fragments from at least two different viruses.  
     
     
         24 . The recombinant adenovirus of  claim 23 , wherein at least one of said at least two different viruses is an adenovirus.  
     
     
         25 . The recombinant adenovirus of  claim 24 , wherein at least one of said protein fragments comprises a tissue tropism-determining fragment of a fiber protein of a subgroup B adenovirus origin.  
     
     
         26 . A cell for producing a recombinant adenovirus having a tissue tropism for smooth muscle cells, said cell comprising: 
 means for assembling said recombinant adenovirus;    wherein said means includes at least one adenoviral nucleic acid encoding an adenoviral fiber protein having at least a tissue tropism-determining fragment of a subgroup B adenoviral fiber protein.    
     
     
         27 . The cell of  claim 26 , wherein said cell is or is derived from a PER.C6 cell (ECACC deposit number 96022940).  
     
     
         28 . An adenovirus capsid having a tissue tropism for smooth muscle cells and/or endothelial cells wherein said adenovirus capsid comprises proteins from at least two different adenoviruses and wherein at least a tissue tropism-determining fragment of a fiber protein is derived from a subgroup B adenovirus.  
     
     
         29 . An adenovirus capsid having a reduced tropism for liver cells as compared to the corresponding wild type adenovirus capsid, said adenovirus capsid comprising: 
 a fiber protein of an adenovirus of subgroup C origin; and    wherein at least a knob domain of the fiber protein is of an adenovirus origin selected from the group consisting of adenovirus 12, adenovirus 16, adenovirus 28, and adenovirus 40-L.    
     
     
         30 . A method of delivering nucleic acid to cells selected from the group of cells consisting of smooth muscle cells, endothelial cells and both smooth muscle and endothelial cells, said method comprising: 
 administering to said cells an adenovirus capsid comprising proteins from at least two different adenoviruses,    wherein at least a tissue tropism-determining fragment of a fiber protein is derived from a subgroup B adenovirus.    
     
     
         31 . A recombinant adenovirus of a subgroup C origin having an increased tropism for smooth muscle cells when compared to an adenovirus of serotype 5 comprising: 
 a recombinant adenovirus capsid comprising a fiber protein, wherein at least a knob domain of the fiber protein is of an adenovirus of a subgroup B origin;    wherein said adenovirus of subgroup B origin is selected from the group consisting of adenovirus 11, adenovirus 16, adenovirus 35, and adenovirus 51.    
     
     
         32 . The recombinant adenovirus of  claim 31 , wherein at least one of said at least two different viruses is an adenovirus.  
     
     
         33 . The recombinant adenovirus of  claim 31 , wherein said subgroup B adenovirus is adenovirus 16.  
     
     
         34 . The recombinant adenovirus of  claim 31 , wherein at least one of said peptides is of adenovirus subgroup C origin.  
     
     
         35 . The recombinant adenovirus of  claim 31 , further comprising an adenoviral nucleic acid incorporated within a genome of said recombinant adenovirus.  
     
     
         36 . The recombinant adenovirus of  claim 35 , wherein said adenoviral nucleic acid comprises sequences from at least two different adenoviruses.  
     
     
         37 . The recombinant adenovirus of  claim 35 , wherein said adenoviral nucleic acid comprises at least one sequence encoding a fiber protein having a tissue tropism-determining fragment of a subgroup B adenovirus fiber protein.  
     
     
         38 . The recombinant adenovirus of  claim 35 , wherein said adenoviral nucleic acid is modified such that the capacity of said adenoviral nucleic acid to replicate in a target cell has been reduced or disabled.  
     
     
         39 . The recombinant adenovirus of  claim 31 , wherein said recombinant adenovirus is a minimal adenovirus vector or an Ad/AAV chimeric vector.  
     
     
         40 . The recombinant adenovirus of  claim 31 , further comprising at least one non-adenoviral nucleic acid incorporated within a genome of said recombinant adenovirus.  
     
     
         41 . The recombinant adenovirus of  claim 40 , wherein at least one of said non-adenoviral nucleic acids is a gene encoding a protein selected from the group of proteins consisting of: an apolipoprotein, a nitric oxide synthase, a herpes simplex virus thymidine kinase, an interleukin-3, an interleukin-1α, an angiogenesis protein, an anti-angiogenesis protein, an anti-proliferation protein, a smooth muscle cell anti-migration protein, a vascular endothelial growth factor, a basic fibroblast growth factor, a hypoxia inducible factor 1α and a PAI-1.  
     
     
         42 . A recombinant adenovirus capsid having an increased tropism for smooth muscle cells as compared to the corresponding wild type adenovirus, said recombinant adenovirus capsid comprising: 
 a chimeric fiber protein, wherein at least a knob domain of the chimeric fiber protein is of an adenovirus of subgroup B origin; and    wherein the adenovirus of subgroup B origin is selected from the group consisting of adenovirus 11, adenovirus 16, adenovirus 35, and adenovirus 51;    wherein a remaining part of the chimeric fiber protein is of an adenovirus of a subgroup C origin.    
     
     
         43 . A recombinant adenovirus having a capsid with an increased tropism for smooth muscle cells as compared to the corresponding wild type adenovirus, said recombinant adenovirus comprising: 
 a chimeric fiber protein comprising at least the knob domain of a fiber protein of an adenovirus selected from the group consisting of adenovirus 11, adenovirus 16, adenovirus 35, and adenovirus 51;    wherein a remaining part of the chimeric fiber protein is of an adenovirus of a subgroup C origin.    
     
     
         44 . The recombinant adenovirus of  claim 43 , further comprising an adenoviral nucleic acid incorporated within a genome of said recombinant adenovirus.  
     
     
         45 . The recombinant adenovirus of  claim 44 , wherein said adenoviral nucleic acid comprises a sequence encoding the chimeric fiber protein.  
     
     
         46 . The recombinant adenovirus of  claim 43 , wherein said recombinant adenovirus is a minimal adenovirus or an Ad/AAV chimeric vector.  
     
     
         47 . The recombinant adenovirus of  claim 43 , wherein said different adenovirus serotype is an adenovirus serotype of subgroup C.  
     
     
         48 . The recombinant adenovirus of  claim 43 , wherein said adenovirus of subgroup C origin is adenovirus serotype 5.  
     
     
         49 . A cell for producing a recombinant adenovirus having a tissue tropism for smooth muscle cells, said cell comprising: 
 means for assembling the recombinant adenovirus;    wherein said means comprises at least one adenoviral nucleic acid encoding a chimeric adenoviral fiber protein having at least a knob domain of a fiber protein of adenovirus serotype 16; and wherein the remaining part of the fiber protein is of a different adenovirus serotype; and    wherein said cell is of a PER.C6 cell (ECACC deposit number 96022940) origin.    
     
     
         50 . The cell of  claim 49 , wherein said different adenovirus serotype is an adenovirus of subgoup C.  
     
     
         51 . The cell of  claim 50 , wherein said adenovirus of subgroup C is adenovirus serotype 5.  
     
     
         52 . A recombinant adenovirus capsid having a reduced tropism for liver cells as compared to the corresponding wild type adenovirus, said recombinant adenovirus comprising: 
 a chimeric fiber protein comprising at least the knob domain of a fiber protein of adenovirus serotype 16;    wherein the remaining part of the fiber protein is of an adenovirus of a subgroup C origin.    
     
     
         53 . The adenovirus capsid of  claim 52 , wherein said different adenovirus serotype is an adenovirus serotype of subgroup C.  
     
     
         54 . The adenovirus capsid of  claim 52 , wherein said adenovirus of subgroup C origin is adenovirus serotype 5.