Recombinant clusterin and use thereof in the treatment and prevention of disease

Abstract

Recombinant clusterin polypeptides and compositions comprising the same are provided. In some aspects, recombinant clusterin or nucleic acids encoding the same may be used for treating and preventing an abnormality of morphology and function in a mammal with disease (e.g., cardiovascular diseases or alcoholism).

1 . (canceled) 
     
     
         2 . The method of  claim 47 , comprising a Clusterin coding sequence having a deletion of the nuclear localization signal and/or transmembrane domain. 
     
     
         3 . The method of  claim 47 , comprising a Clusterin coding sequence and a heterologous polypeptide sequence fused to said Clusterin coding sequence. 
     
     
         4 . The method of  claim 3 , wherein the heterologous polypeptide sequence comprises a protease cleavage site. 
     
     
         5 . The method of  claim 3 , wherein the protease cleavage site is a thrombin cleavage site. 
     
     
         6 . The method of  claim 47 , wherein the polypeptide is a fusion protein comprising the Clusterin coding sequence and a heterologous polypeptide sequence. 
     
     
         7 . The method of  claim 47 , wherein the polypeptide is aglycosylated. 
     
     
         8 . The method of  claim 47 , wherein the Clusterin coding sequence has a deletion of the nuclear localization signal. 
     
     
         9 . The method of  claim 47 , wherein the Clusterin coding sequence has a deletion of the transmembrane domain. 
     
     
         10 . The method of  claim 9 , wherein the deletion of the transmembrane domain disrupts endoplasmic reticulum (ER)-targeting of the polypeptide. 
     
     
         11 . The method of  claim 47 , further comprising a tag sequence. 
     
     
         12 . The method of  claim 11 , further comprising a protease cleavage site positioned between the tag sequence and the clusterin coding sequence. 
     
     
         13 . The method of  claim 11 , wherein the protease cleavage site is a thrombin or enteropeptidase cleavage site. 
     
     
         14 . The method of  claim 11 , wherein the tag sequence is a polyhistidine tag. 
     
     
         15 . The method of  claim 11 , wherein the tag sequence is positioned N-terminally relative to the Clusterin coding sequence. 
     
     
         16 . The method of  claim 11 , wherein the tag sequence is positioned C-terminally relative to the Clusterin coding sequence. 
     
     
         17 - 46 . (canceled) 
     
     
         47 . A method of treating or preventing alcoholism in a subject comprising administering an effective amount of a composition comprising a polypeptide comprising:
 (i) a Clusterin coding sequence, said coding sequence having a deletion of the nuclear localization signal and/or transmembrane domain; or   (ii) a Clusterin coding sequence and a heterologous polypeptide sequence fused to said Clusterin coding sequence.   
     
     
         48 . The method of  claim 47 , wherein the composition is administered by intravenous injection, intratissue injection and/or catheter delivery. 
     
     
         49 . The method of  claim 47 , wherein the nucleic acid sequence encoding the Clusterin polypeptide is operably linked to a promoter. 
     
     
         50 . The method of  claim 49 , wherein the promoter is a gene promoter functional in mammalian cells. 
     
     
         51 . The method of  claim 50 , wherein the polynucleotide molecule is part of a gene expression vector, such as a viral expression vector.