Atherosclerosis inhibition via modulation of monocyte-macrophage phenotype using apo a-i milano gene transfer

Abstract

A method of changing the phenotype of monocytes and macrophages from a pro inflammatory MI phenotype to an anti-inflammatory M2 phenotype is disclosed. The method can comprises providing a composition comprising a recombinant adeno-associated virus (rAAV) vector comprising an exogenous gene encoding ApoA-1 Milano or a fragment thereof, and administering the composition to mammal in need thereof to change the phenotype of monocytes or macrophages from a pro inflammatory M1 phenotype to an anti inflammatory M2 phenotype. By changing the phenotype of monocytes or macrophages from a pro-inflammatory M1 phenotype to anti-inflammatory M2 phenotype, atherosclerosis can be treated. A method of monitoring macrophage phenotypic switching and a method of assessing the efficacy of the treatment of atherosclerosis are also described.

What is claimed is: 
     
         1 . A method, comprising:
 providing a composition comprising a recombinant adeno-associated virus (rAAV) vector comprising an exogenous gene encoding ApoA-I Milano, or fragment thereof; and   administering the composition to a mammal in need of changing the phenotype of a monocyte or macrophage from a proinflammatory M1 phenotype to an anti-inflammatory M2 phenotype to change the phenotype of the monocyte or macrophage from the proinflammatory M1 phenotype to the anti-inflammatory M2 phenotype.   
     
     
         2 . The method of  claim 1 , further comprising monitoring monocyte or macrophage phenotypic switching in the mammal. 
     
     
         3 . The method of  claim 1 , further comprising assessing the efficacy of a treatment for atherosclerosis in the mammal. 
     
     
         4 . The method of  claim 1 , wherein the rAAV vector is rAAV8 vector comprising an exogenous gene encoding ApoA-I Milano or a fragment thereof. 
     
     
         5 . The method of  claim 4 , wherein the rAAV8 is produced by co-transfecting a host cell with a first plasmid and a second plasmid, wherein the first plasmid genome for the rAAV8 is derived from AAV serotype 2 and the second plasmid is derived from AAV serotypes 2 and 8 (Rep2Cap8). 
     
     
         6 . The method of  claim 1 , wherein the rAAV vector is rAAV2 vector comprising an exogenous gene encoding ApoA-I Milano or a fragment thereof. 
     
     
         7 . The method of  claim 6 , wherein the rAAV2 is produced by co-transfecting a host cell with a first plasmid and a second plasmid, wherein the first plasmid genome for the rAAV2 is derived from AAV serotype 2 and the second plasmid is derived from AAV serotype 2 (Rep2Cap2). 
     
     
         8 . The method of  claim 1 , wherein the monocyte or macrophage is a circulating monocyte or macrophage. 
     
     
         9 . The method of  claim 1 , wherein the monocyte or macrophage is a peritoneal monocyte or macrophage. 
     
     
         10 . The method of  claim 1 , wherein changing the phenotype of the monocyte or macrophage from the proinflammatory M1 phenotype to the anti-inflammatory M2 phenotype inhibits or treats atherosclerosis. 
     
     
         11 . The method of  claim 1 , wherein changing the phenotype of the monocyte or macrophage from the proinflammatory M1 phenotype to the anti-inflammatory M2 phenotype reduces atherosclerosis. 
     
     
         12 . The method of  claim 11 , wherein the atherosclerosis is reduced in a whole aorta, an aortic sinus, an inominate artery or combinations thereof. 
     
     
         13 . The method of  claim 1 , wherein changing the phenotype of the monocyte or macrophage from the proinflammatory M1 phenotype to the anti-inflammatory M2 phenotype reduces plaque lipid content in an aortic sinus, an innominate artery, or both. 
     
     
         14 . The method of  claim 1 , wherein changing the phenotype of the monocyte or macrophage from the proinflammatory M1 phenotype to the anti-inflammatory M2 phenotype reduces plaque macrophage content in an aortic sinus, an innominate artery or both. 
     
     
         15 . The method of  claim 1 , wherein the rAAV vector is produced by
 providing a first plasmid comprising ApoA-I Milano or a fragment thereof;   providing a second plasmid complimentary to the first plasmid and which comprises components for rescue and packaging;   co-transfecting the first plasmid and the second plasmid into a host cell; and   generating a quantity of the rAAV vector from the co-transfected host cell.   
     
     
         16 . The method of  claim 15 , wherein the pair of the first plasmid and the second plasmid is selected such that the rAAV vector is targeted for delivery to a specific tissue type. 
     
     
         17 . The method of  claim 15 , wherein the second plasmid further comprises AAV rescue and packaging components derived from an AAV serotype selected from the group consisting of AAV1, AAV2, AAV5, AAV7, AAV8, AAV9, AAV10 and combinations thereof. 
     
     
         18 . A method, comprising:
 measuring the expression level of one or more markers selected from the group consisting of MCP-1, IL-6, TNF-a, Arg-1, Ym-1 and CD206 to monitor monocyte or macrophage phenotypic switching in a mammal in need thereof; and   determining the presence of a phenotypic switch from a proinflammatory M1 macrophage to an anti-inflammatory M2 macrophage when MCP-1, IL-6 and/or TNF-α is down-regulated, and/or Arg-1, Ym-1 and/or CD206 is up-regulated, or   determining the absence of a phenotypic switch from the proinflammatory M1 macrophage to the anti-inflammatory M2 macrophage when MCP-1, IL-6 and/or TNF-a is not down-regulated, and/or Arg-1, Ym-1 and/or CD206 is not up-regulated, or   determining the presence of a phenotypic switch from an anti-inflammatory M2 macrophage to a proinflammatory M1 macrophage when MCP-1, IL-6 and/or TNF is up-regulated, and/or Arg-1 and/or CD206 is down-regulated, or   determining the presence of a phenotypic switch from a proinflammatory M1 macrophage to an anti-inflammatory M2 macrophage when MCP-1, IL-6 and/or TNF-a is down-regulated, and/or when, Arg-1, Ym-1 and/or CD206 is not up-regulated.   
     
     
         19 . A method for assessing the efficacy of a treatment for atherosclerosis in a mammalian subject in need thereof, comprising:
 determining the phenotype of the macrophages in the mammalian subject; and   making a determination that a treatment is providing beneficial results to a mammalian subject if a phenotypic switch from proinflammatory M1 macrophage to anti-inflammatory M2 macrophage is detected, or   making a determination that a treatment is not providing beneficial results to the mammalian subject if a phenotypic switch from anti-inflammatory M2 macrophage to proinflammatory M1 macrophage is detected.   
     
     
         20 . The method of  claim 19 , wherein the treatment for atherosclerosis is treatment with ApoA-1 Milano.