Recombinant viruses and their use for treatment of atherosclerosis and other forms of coronary artery disease and method, reagent, and kit for evaluating susceptibility 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 an adeno-associated virus comprising a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL), wherein the nucleic acid sequence is operably linked to a 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 an adeno-associated virus comprising a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL) wherein the nucleic acid sequence is operably linked to a 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 an adeno-associated virus comprising a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL), wherein the nucleic acid sequence is operably linked to a 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 an adeno-associated virus comprising a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL), wherein the nucleic acid sequence is operably linked to a 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 an adeno-associated virus comprising a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL) wherein the nucleic acid sequence is operably linked to a 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 an adeno-associated virus comprising a nucleic acid sequence coding for a biologically active human lipoprotein lipase (LPL), wherein the nucleic acid sequence is operably linked to a 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 . The method according to one of  claims 16 - 21 , wherein the promoter is a viral promoter. 
     
     
         24 . The method according to  claim 23 , wherein the viral promoter is selected from the group consisting of an E1A promoter, a MLP promoter, a CMV promoter, and a RSV LTR promoter. 
     
     
         25 . The method according to  claim 24 , wherein the viral promoter is the RSV LTR promoter. 
     
     
         26 . The method according to  claim 22 , wherein the promoter is a viral promoter. 
     
     
         27 . The method according to  claim 22 , wherein the viral promoter is selected from the group consisting of an E1A promoter, a MLP promoter, a CMV promoter, and a RSV LTR promoter. 
     
     
         28 . The method according to  claim 22 , wherein the viral promoter is the RSV LTR promoter. 
     
     
         29 . 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 an adeno-associated virus comprising a nucleic acid sequence coding for a replacement lipoprotein lipase enzyme, 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. 
     
     
         30 . The method according to  claim 29 , wherein the adeno-associated virus further comprises a promoter effective to promote expression of the nucleic acid sequence in human cells. 
     
     
         31 . The method according to  claim 29 , wherein the adeno-associated virus is administered by parenteral injection. 
     
     
         32 . An adeno-associated virus comprising a nucleic acid sequence coding for a biologically active lipoprotein lipase (LPL). 
     
     
         33 . The adeno-associated virus according to  claim 32 , wherein the nucleic acid sequence is placed under the control of a promoter permitting its expression in an infected cell. 
     
     
         34 . The adeno-associated virus according to  claim 32 , wherein the nucleic acid sequence is a cDNA sequence. 
     
     
         35 . The adeno-associated virus according to  claim 32 , wherein the nucleic acid sequence codes for human LPL. 
     
     
         36 . The adeno-associated virus according to  claim 32 , wherein the promoter is selected from the group consisting of E1A, MLP, CMV, and RSV LTR promoters. 
     
     
         37 . An adeno-associated virus comprising a cDNA sequence coding for a biologically active lipoprotein lipase (LPL) under the control of an RSV LTR promoter. 
     
     
         38 . The adeno-associated virus according to  claim 32 , 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. 
     
     
         39 . The adeno-associated virus according to  claim 38 , wherein the secretion sequence is the native secretion sequence of lipoprotein lipase. 
     
     
         40 . The adeno-associated virus according to  claim 32 , wherein the adeno-associated virus lacks the rep gene. 
     
     
         41 . The adeno-associated virus according to  claim 32 , wherein the adeno-associated virus lacks the cap gene. 
     
     
         42 . The adeno-associated virus according to  claim 32 , wherein the adeno-associated virus lacks the rep and cap genes. 
     
     
         43 . The adeno-associated virus according to  claim 32 , further comprising a gene coding for an apolipoprotein. 
     
     
         44 . The adeno-associated virus according to  claim 43 , wherein the apolipoprotein is selected from the group consisting of ApoA-I and ApoA-IV. 
     
     
         45 . An adeno-associated virus comprising a nucleic acid sequence coding for hepatic lipase and further comprising a nucleic acid sequence coding for apolipoprotein selected from the group consisting of ApoA-I and ApoA-IV, wherein the nucleic acid sequences are placed under the control of a promoter permitting their expression in an infected cell. 
     
     
         46 . A composition comprising the adeno-associated virus according to  claim 32  and a pharmaceutically acceptable vehicle. 
     
     
         47 . The composition according to  claim 46 , wherein the composition is in an injectable form. 
     
     
         48 . A mammalian cell infected in vitro with the adeno-associated virus according to  claim 32 , whereby a biologically active lipoprotein lipase is expressed from the adeno-associated virus. 
     
     
         49 . The mammalian cell according to  claim 48 , wherein the cell is a human cell. 
     
     
         50 . The mammalian cell according to  claim 49 , wherein the cell is selected from the group consisting of a fibroblast, myoblast, hepatocyte, endothelial cell, glial cell, and keratinoctyte. 
     
     
         51 . A composition comprising the infected mammalian cell according to  claim 48  and an extracellular matrix. 
     
     
         52 . The composition according to  claim 51 , wherein the extracellular matrix comprises a gelling compound selected from the group consisting of collagen, gelatin, glycosaminoglycans, fibronectin, and lectins. 
     
     
         53 . The composition according to  claim 52 , wherein the extracellular matrix comprises a support permitting anchorage of the infected cell. 
     
     
         54 . The composition according to  claim 53 , wherein the support comprises polytetrafluoroethylene fibers. 
     
     
         55 . The adeno-associated virus of  claim 32 , wherein the adeno-associated virus is defective.