Method for PR-39 peptide mediated selective inhibition of IkappaBalpha degradation

Abstract

The present invention provides both a method and means for regulating IκBα degradation, NFκB activity, and NFκB-dependent gene expression within living cells, tissues, and organs in-situ. The selective regulation is performed using native PR-39 peptide or one of its shorter-length homologs, for interaction with such IκBα and proteasomes as are present in the cytoplasm of viable cells. The result of PR-39 peptide interaction with IκBα is a selective alteration in the intracellular proteolytic activity of proteasomes, which in turn, causes a reduction of IκBα, a decrease of NFκB activity, and a down-regulation of NFκB-dependent expression.

1 - 2 . (canceled)  
     
     
         3 . A method for selective control of NFκB-dependent gene expression in-situ within a collection of viable cells, said method comprising the steps of: 
 identifying a collection of cells comprising viable cells in-situ as a target for controlling NFκB-dependent gene expression;    providing means for effecting an introduction of at least one member selected from the group consisting of the PR-39 oligopeptide collective to the cytoplasm of said targeted collection of cells;    introducing at least one member of the PR-39 oligopeptide collective to the cytoplasm of said targeted collection of cells using said effecting means;    allowing said introduced PR-39 oligopeptide collective member to interact with such IκBα and proteasomes as are present within the cytoplasm of said targeted collection of cells whereby    (a) at least some of the proteasomes interact with the PR-39 oligopeptide collective member,    (b) at least a part of the proteolytic activity mediated by said proteasomes becomes selectively altered by said interaction,    (c) the selectively altered proteolytic activity of said proteasomes results in a marked reduction of IκBα degradation in-situ within the cytoplasm of said targeted collection of cells,    (d) said reduction of IκBα degradation results in a decrease in activity for such NFκB transcription factor as is present intracellularly, and    (e) NFκB-dependent gene expression is selectively controlled in said targeted collection of viable cells.    
     
     
         4 . The method as recited in  claim 3  wherein said collection of viable cells includes at least one type of cell selected from the group consisting of endothelial cells, myocytes and myoblasts, fibrocytes and fibroblasts, epithelial cells, osteocytes and osteoblasts, neuronal cells and glial cells, erythrocuctes, leukocytes, and progenitor cells of all types.  
     
     
         5 . The method as recited in  claim 3  wherein said collection of cells comprises at least one tissue selected from the group consisting of myocardium, skeletal muscle, smooth muscle, an artery, a vein, lung, brain, kidney, spleen, liver, gastrointestinal tissue, nerve tissue, limbs, and extremities.  
     
     
         6 . The method as recited in  claim 3  wherein the means for an introduction of a PR-39 oligopeptide collective member include one selected from the group consisting of catheter-based introduction means, injection-based introduction means, infusion-based introduction means, localized intravascular introduction means, liposome-based introduction means, receptor-specific peptide introduction means, slow releasing means for peptide secretion in living cells and sequested organisms.  
     
     
         7 . The method as recited in  claim 3  wherein the means for an introduction of a PR-39 oligopeptide collective member includes the DNA sequences coding for PR-39 oligopeptides of different sizes inserted in a suitable vector for transfection and subsequent expression of peptides within said cells.  
     
     
         8 . The method as recited in  claim 3  wherein said method is practiced under in-vivo conditions.  
     
     
         9 . The method as recited in  claim 3  wherein said method is practiced under in-vitro conditions.  
     
     
         10 - 13 . (canceled)