Somatic gene therapy to suppress secondary cataract formation following eye surgery

Abstract

Disclosed is a replication-recombinant virus, preferably an adenovirus that lacks E1a, E1b and E4 ORF 6, capable of infecting an eye and comprising a lens epithelial cell specific promoter driving an ORF encoding at least one protein, which when expressed in lens epithelial cells of an eye suppresses, at the level of the germinative epithelium of the lens of the eye, cellular proliferation which is stimulated by eye surgery and which would otherwise result in secondary cataract formation in the eye. Also disclosed is the use of the recombinant virus for the treatment of an eye, undergoing eye (e.g. cataract) surgery, in order to reduce the incidence of cellular proliferation in the eye following the surgery and thereby to prevent the formation of secondary cataracts.

What is claimed is:  
     
         18 . (New) A replication-defective recombinant virus for infecting lens epithelial cells of an eye, said virus comprising: 
 (a) an open reading frame (ORF) encoding a protein that, upon expression in a lens epithelial cell, suppresses cellular proliferation that is stimulated by eye surgery; and    (b) a promoter comprising nucleotide sequences derived from at least one promoter that is active specifically in human lens epithelial cells,    wherein the promoter is functionally linked to the ORF and expresses a biologically active amount of the protein encoded by the ORF.    
     
     
         19 . (New) The replication-defective recombinant virus of  claim 18  which is an adenovirus.  
     
     
         20 . (New) The replication-defective recombinant virus of  claim 19 , wherein said virus lacks E1a, E1b, and E4 ORF 6.  
     
     
         21 . (New) The replication-defective recombinant virus of  claim 18  wherein the promoter sequence is one of the following: 
 a) a promoter of a human Major Intrinsic Protein gene or  
 b) a promoter of a human β A3/A1-crystallin gene or  
 c) a composite promoter comprising the MIP promoter or the β-crystallin promoter, in combination with elements of an early growth response gene promoter.  
 
     
     
         22 . (New) The replication-defective recombinant virus of  claim 21  wherein the promoter sequence is one of the following: 
 a) a promoter comprising the portion of the human Major Intrinsic Protein gene from −259 nt to +34 nt of SEQ ID NO:2, or  
 b) a promoter comprising the portion of the human β A3/A1-crystallin gene from −345 nt to +45 nt of SEQ ID NO:4, or  
 c) a composite promoter comprising 
 i) at least one element chosen from the group consisting of: the promoter of the human Major Intrinsic Protein gene, the portion of the promoter of the human Major Intrinsic Protein gene from −259 nt to −34 nt of SEQ ID NO:2, the β-crystallin promoter, and the portion of the human β-crystallin gene from −345 nt to +45 nt of SEQ ID NO:4; in combination with  
 ii) the portion of the promoter of the rat Early Growth Response-1 gene from −518 nt to −236 nt of SEQ ID NO:1.  
 
 
     
     
         23 . (New) The replication-defective recombinant virus of  claim 18 , wherein the promoter sequence is non-methylated.  
     
     
         24 . (New) A method for the treatment of an eye of a patient, undergoing eye surgery, in order to reduce the incidence of cellular proliferation in the eye following the eye surgery, and thereby prevent the formation of secondary cataracts, comprising the step of: 
 treating the eye with a replication-defective recombinant virus of  claim 18 .    
     
     
         25 . (New) The method of  claim 24  wherein the eye is treated during the eye surgery with the recombinant virus.  
     
     
         26 . (New) The method of  claim 25 , comprising the additional step of treating the eye at least one more time with the recombinant virus after the surgery.  
     
     
         27 . (New) The method of  claim 24 , comprising the additional step of treating the patient with acyclovir or a nucleoside analogue structurally related to acyclovir when the recombinant virus contains a thymidine kinase ORF of a Herpes virus or the UL97 ORF of a Cytomegalovirus.  
     
     
         28 . (New) A composition for the treatment of an eye, undergoing eye surgery, in order to reduce the incidence of cellular proliferation of the lens epithelial cells in the eye following surgery and thereby prevent secondary cataracts, comprising the replication-defective recombinant virus of  claim 18 .  
     
     
         29 . (New) A replication-defective recombinant adenovirus virus comprising an ORF selected from the group consisting of: 
 a) a non-phosphorylatable retinoblastoma ORF;    b) a dominant negative mutant of a RAS ORF; or    c) a thymidine kinase ORF of a Herpes virus;    wherein 
 i) said ORF is under the control of a promoter sequence comprising nucleotide sequences derived from at least one promoter that is active specifically in human lens epithelial cells;  
 ii) said replication-defective recombinant virus is capable of infecting lens epithelial cells of an open lens capsule of an eye.  
   
     
     
         30 . (New) The replication-defective recombinant adenovirus of  claim 29  which lacks E1a, E1b and E4 ORF 6.  
     
     
         31 . (New) The replication-defective recombinant virus of  claim 29  wherein the promoter sequence is one of the following: 
 a) the promoter of a human Major Intrinsic Protein gene in SEQ ID NO:2; or  
 b) the promoter of a human β A3/A1-crystallin gene in SEQ ID NO:4.  
 
     
     
         32 . (New) The replication-defective recombinant adenovirus of  claim 31  which lacks E1a, E1b and E4 ORF 6.  
     
     
         33 . (New) A method for the treatment of an eye of a patient, undergoing eye surgery, in order to reduce the incidence of cellular proliferation in the eye following the eye surgery, and thereby prevent the formation of secondary cataracts, comprising the step of treating the eye with a replication-defective recombinant adenovirus virus of  claim 29 .  
     
     
         34 . (New) A method for the treatment of an eye of a patient, undergoing eye surgery, in order to reduce the incidence of cellular proliferation in the eye following the eye surgery, and thereby prevent the formation of secondary cataracts, comprising the step of treating the eye with a replication-defective recombinant adenovirus virus of  claim 32 .  
     
     
         35 . (New) The replication-defective recombinant virus of  claim 18 , wherein the promoter sequence is a composite promoter sequence comprising a basal transcription region and at least one transcription enhancer element, the enhancer including but not limited to an immediate early enhancer.  
     
     
         36 . (New) The replication-defective recombinant virus of  claim 29 , wherein the promoter sequence is a composite promoter sequence comprising a basal transcription region and at least one transcription enhancer element, the enhancer including but not limited to an immediate early enhancer.