Recombinant Viruses and their Use for Treatment of Atherosclerosis and Othe Forms of Coronary Artery Disease and Method, Reagent, and Kit for Evaluating Susceptibiity to Same

Abstract

Recombinant viruses comprising a heterologous DNA sequence coding for a lipase involved in lipoprotein metabolism. The invention also concerns the preparation and use in therapy of said recombinant viruses, especially for the treatment or prevention of dyslipoproteinemia-related pathologies.

1 - 15 . (canceled) 
     
     
         16 . A method for treating a patient with dyslipoproteinaemia comprising administering to the patient via intravenous injection a recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL), wherein the nucleic acid sequence is operably linked to a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter, and wherein the nucleic acid sequence is expressed so as to cause a reduction of lipoprotein in the patient. 
     
     
         17 . A method for treating a patient with hypertriglyceridaemia comprising administering to the patient via intravenous injection a recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL) wherein the nucleic acid sequence is operably linked to a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter, and wherein the nucleic acid sequence is expressed so as to cause a reduction of triglyceride in the patient. 
     
     
         18 . A method for treating a patient with hypercholesterolaemia comprising administering to the patient via intravenous injection a recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL), wherein the nucleic acid sequence is operably linked to a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter, and wherein the nucleic acid sequence is expressed so as to cause a reduction of cholesterol in the patient. 
     
     
         19 . A method for treating a patient with hyperlipidaemia comprising administering to the patient via intravenous injection a recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL), wherein the nucleic acid sequence is operably linked to a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter, and wherein the nucleic acid sequence is expressed so as to cause a reduction of lipid in the patient. 
     
     
         20 . A method for treating a patient with familial hypertriglyceridaemia comprising administering to the patient via intravenous injection a recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL) wherein the nucleic acid sequence is operably linked to a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter, and wherein the nucleic acid sequence is expressed so as to cause a reduction of triglyceride in the patient. 
     
     
         21 . A method for treating a patient with combined familial hyperlipidaemia and postprandial hyperlipidaemia comprising administering to the patient via intravenous injection a recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL), wherein the nucleic acid sequence is operably linked to a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter, and wherein the nucleic acid sequence is expressed so as to cause a reduction of lipid in the patient. 
     
     
         22 . The method of treatment according to  claim 16 , wherein the adeno-associated virus is administered by direct injection into the patient's portal vein, such that viral infection is targeted to the liver. 
     
     
         23 . A method for preventing or delaying the onset of coronary artery disease in a human individual having lipoprotein lipase enzyme in which a serine residue is present at amino acid 291 in the enzyme, comprising administering to the individual a recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a nucleic acid sequence coding for a replacement lipoprotein lipase enzyme under the control of a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter, said replacement lipoprotein lipase enzyme having an asparagine residue as amino acid 291, wherein the replacement lipoprotein lipase enzyme is produced in the individual to provide a functional lipoprotein lipase enzyme. 
     
     
         24 . The method according to  claim 23 , wherein the adeno-associated virus further comprises a promoter effective to promote expression of the nucleic acid sequence in human cells. 
     
     
         25 . The method according to  claim 23 , wherein the adeno-associated virus is administered by parenteral injection. 
     
     
         26 . A recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a nucleic acid sequence coding for a biologically active lipoprotein lipase (LPL) under the control of a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter. 
     
     
         27 . The adeno-associated virus according to  claim 26 , wherein the nucleic acid sequence is placed under the control of a CMV promoter. 
     
     
         28 . The adeno-associated virus according to  claim 26 , wherein the nucleic acid sequence is a cDNA sequence. 
     
     
         29 . The adeno-associated virus according to  claim 26 , wherein the nucleic acid sequence codes for human LPL. 
     
     
         30 . The adeno-associated virus according to  claim 26 , wherein the nucleic acid sequence is placed under the control of an RSV LTR promoter. 
     
     
         31 . A recombinant defective adeno-associated virus, wherein the rep gene, the cap gene, or the rep and cap genes of the adeno-associated virus are deleted and replaced by a cDNA sequence coding for a biologically active lipoprotein lipase (LPL) under the control of a cytomegalovirus (CMV) or Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter. 
     
     
         32 . The adeno-associated virus according to  claim 26 , wherein the virus further comprises a nucleic acid sequence enabling the lipoprotein lipase to be directed into a pathway of secretion in an infected cell. 
     
     
         33 . The adeno-associated virus according to  claim 31 , wherein the secretion sequence is the native secretion sequence of lipoprotein lipase. 
     
     
         34 . The adeno-associated virus according to  claim 26 , wherein the adeno-associated virus lacks the rep gene. 
     
     
         35 . The adeno-associated virus according to  claim 26 , wherein the adeno-associated virus lacks the cap gene. 
     
     
         36 . The adeno-associated virus according to  claim 26 , wherein the adeno-associated virus lacks the rep and cap genes. 
     
     
         37 . A composition comprising the adeno-associated virus according to  claim 26  and a pharmaceutically acceptable vehicle. 
     
     
         38 . The composition according to  claim 36 , wherein the composition is in an injectable form. 
     
     
         39 . A mammalian cell infected in vitro with the adeno-associated virus according to  claim 26 , whereby a biologically active lipoprotein lipase is expressed from the adeno-associated virus. 
     
     
         40 . The mammalian cell according to  claim 38 , wherein the cell is a human cell. 
     
     
         41 . The mammalian cell according to  claim 39 , wherein the cell is selected from the group consisting of a fibroblast, myoblast, hepatocyte, endothelial cell, glial cell, and keratinoctyte. 
     
     
         42 . A composition comprising the infected mammalian cell according to  claim 38  and an extracellular matrix. 
     
     
         43 . The composition according to  claim 41 , wherein the extracellular matrix comprises a gelling compound selected from the group consisting of collagen, gelatin, glycosaminoglycans, fibronectin, and lectins. 
     
     
         44 . The composition according to  claim 42 , wherein the extracellular matrix comprises a support permitting anchorage of the infected cell. 
     
     
         45 . The composition according to  claim 43 , wherein the support comprises polytetrafluoroethylene fibers.