Composition and method for clusterin-mediated stem cell therapy for treatment of atherosclerosis and heart failure

Abstract

Methods and compositions are disclosed for inhibiting, deterring or preventing apoptosis of cardiac myocytes, transplanted stem cells, vascular stem cells, and vascular smooth muscle cells by means of expressing or synthesizing clusterin. Also disclosed are methods and compositions for producing recombinant clusterin, or its biologically active peptides, and for induction of clusterin-associated lipoproteins or enzymes for deterring or preventing inflammatory injury and apoptosis induced by oxLDL, oxysterols, cytokines, and Fas Ligand. Also disclosed is an induction method and composition for enhancing expression of ALDH and ALDH-associated enzymes or co-factors to prevent cytotoxicity or detoxification. Therapeutic methods providing new expression or overexpression of clusterin in vascular or cardiac tissue are expected to inhibit the formation of atherosclerotic lesions, stabilize existing atherosclerotic plaques, and repair failing or damaged cardiac tissue.

1 - 10 . (canceled) 
     
     
         11 . A method of treating or preventing atherosclerosis, or a complication thereof, in a mammal, comprising:
 transplanting an isolated population of transfected autologous or allogenic bone marrow stromal cells into cardiac tissue of a mammal by direct delivery of the cells to a cardiac site where expression of clusterin is desired, wherein said cardiac site comprises an atherosclerotic lesion or an area that is at risk of forming an atherosclerotic lesion, wherein said stromal cells comprise bone marrow stem cells, and wherein said transfected cells contain a nucleic acid encoding clusterin protein, or encoding a truncated clusterin protein lacking the transmembrane domain, operably linked to a promoter and capable of being expressed in the transfected cells; and   expressing said clusterin protein encoded by said nucleic acid in an amount sufficient to inhibit apoptosis in said transfected cells and/or adjacent cells, to prevent, or reduce the risk of, formation or rupture of an atherosclerotic lesion.   
     
     
         12 . (canceled) 
     
     
         13 . The method of  claim 11  wherein said amount of clusterin is effective to induce an aldehyde dehydrogenase enzyme that acts as a detoxification agent for oxidized lipoproteins and oxysterols. 
     
     
         14 . The method of  claim 11  wherein said atherosclerotic lesion comprises an aneurism. 
     
     
         15 . The method of  claim 11  wherein said atherosclerotic lesion comprises an unstable plaque caused by hyperlipidemia and wherein said amount of clusterin protein is effective to stabilize said plaque. 
     
     
         16 - 17 . (canceled) 
     
     
         18 . A method of treating heart failure in a mammal comprising:
 transplanting an isolated population of transfected autologous or allogenic bone marrow stromal cells into cardiac tissue of a mammal by direct delivery of the cells to a cardiac site where expression of clusterin is desired, wherein said cardiac site comprises an atherosclerotic lesion or an area that is at risk of forming an atherosclerotic lesion, wherein said stromal cells comprise bone marrow stem cells, and wherein said transfected cells contain a nucleic acid encoding clusterin protein, or encoding a truncated clusterin protein lacking the transmembrane domain, operably linked to a promoter and capable of being expressed in the transfected cells; and   expressing said recombinant clusterin or said truncated clusterin protein lacking the transmembrane domain encoded by said nucleic acid in an amount sufficient to improve heart function and protect said transplanted cells or their progeny from inflammatory injury.   
     
     
         19 . A transgenic mouse comprising, stably integrated into the genome of said mouse, a transgenic DNA sequence encoding clusterin or a truncated clusterin protein lacking the transmembrane domain, operably linked to a promoter, wherein said DNA sequence is expressed, and, as a result of said expression, the transgenic mouse has an increased level of serum clusterin relative to the serum clusterin level in a mouse that does not express said transgenic DNA sequence. 
     
     
         20 . The transgenic mouse of  claim 19  wherein serum from said transgenic mouse has increased oxysterol-binding activity relative to the oxysterol-binding activity of serum from a mouse that does not express said transgenic DNA sequence. 
     
     
         21 . The transgenic mouse of  claim 19  wherein the vascular cells of said transgenic mouse have reduced risk of atherosclerotic lesion formation relative to that of a mouse that does not express said transgenic DNA sequence. 
     
     
         22 . The method of  claim 11 , wherein said cardiac site is exposed to at least one inflammatory agent selected from the group consisting of oxidized low density lipoprotein (oxLDL), oxysterols, cytokines and Fas ligand. 
     
     
         23 . The method of  claim 11  wherein some of the bone marrow stem cells are uncommitted and capable of differentiating into a cardiac myocyte phenotype or cardiac cells. 
     
     
         24 . Recombinantly produced clusterin or truncated clusterin protein lacking the transmembrane domain. 
     
     
         25 . A transgenic stem cell expressing the recombinantly produced clusterin or truncated clusterin protein lacking the transmembrane domain of  claim 24 . 
     
     
         26 . A composition comprising transgenic bone marrow stromal cells including transgenic stem cells according to  claim 25  expressing recombinant clusterin or truncated clusterin protein lacking the transmembrane domain.