Composition and Method for Treatment and Prevention of Restenosis

Abstract

Compositions and methods are disclosed which employ PARIS proteins that are useful for suppressing proliferation of smooth muscle cells. Preferred PARISs are soluble proteins that are secreted by vascular smooth muscle cells, and include PARIS-1 (neuronal pentraxin 1), PARIS-2 (SBP (MIC-1, GDF-15), PARIS-3 (BTG2) and PARIS-4 (soluble fractalkine). Methods of preventing or treating restenosis by administering the new compositions are disclosed. Also disclosed are methods for treating patients undergoing angioplasty procedures, patients with atherosclerosis, and patients with other proliferative disorders, in order to suppress the growth of vascular smooth muscle cells or other cells that play a role in the particular proliferative disorder or condition. A method of screening mRNAs and identifying genes encoding PARISs is also disclosed.

1 . An inhibitor of smooth muscle cell proliferation comprising at least one protein or polypeptide chosen from the group consisting of: 
 PARIS-1, PARIS-2 and PARIS-3,    a biologically active portion of PARIS-1, PARIS-2, PARIS-3 or PARIS-4 having activity for directly or indirectly inhibiting smooth muscle cell proliferation, and    a biologically active addition, deletion or substitution homolog having at least 24% amino acid identity to SEQ. ID NO:1, SEQ. ID NO:2, SEQ. ID NO:3 or SEQ ID NO:4 and having activity for directly or indirectly inhibiting smooth muscle cell proliferation.    
     
     
         2 . A composition for treating a cell proliferation disorder comprising: 
 an inhibitor of smooth muscle cell proliferation comprising at least one protein or polypeptide chosen from the group consisting of:    PARIS-1, PARIS-2 and PARIS-3,    biologically active portions of PARIS-1, PARIS-2, PARIS-3 or PARIS-4, wherein biological activity comprises inhibition of smooth muscle cell proliferation, and    polypeptides having at least 24% amino acid identity to SEQ. ID NO:1, SEQ. ID NO:2, SEQ. ID NO:3 or SEQ ID NO:4; and    optionally, a carrier.    
     
     
         3 . The composition of  claim 2  further comprising PARIS-4.  
     
     
         4 . The composition of  claim 2  wherein said protein or polypeptide has at least 40% amino acid identity with SEQ. ID NO:1, SEQ. ID NO:2 and SEQ. ID NO:3.  
     
     
         5 . A method of inhibiting smooth muscle cell proliferation comprising contacting said cell with the composition of  claim 2 , whereby proliferation of said smooth muscle cell is inhibited.  
     
     
         6 . The method of  claim 5  wherein said smooth muscle cell is a vascular smooth muscle cell.  
     
     
         7 . The method of  claim 6 , wherein said contacting comprises administering to a site at risk of undesired smooth muscle cell proliferation a cell growth inhibitory amount of said composition, whereby a smooth muscle cell proliferative disorder is deterred or prevented.  
     
     
         8 . The method of  claim 6 , wherein said contacting comprises administering to a patient at risk of restenosis an effective amount of said inhibitor to inhibit vascular smooth muscle cell proliferation resulting in deterrence or prevention of restenosis.  
     
     
         9 . The method of  claim 8  wherein said patient is undergoing an angioplasty procedure and said administering comprises administering an effective amount of said inhibitor to said patient before, during or after an angioplasty procedure to deter or prevent restenosis.  
     
     
         10 . The method of  claim 9  wherein said administering includes delivering said inhibitor to an angioplasty site in said patient.  
     
     
         11 . The method of  claim 9  wherein said angioplasty procedure includes placement of a stent at an angioplasty site in said patient.  
     
     
         12 . The method of  claim 11  wherein said stent is a drug-eluting stent capable of releasing said inhibitor in situ.  
     
     
         13 . The method of  claim 10  wherein said contacting comprises administering said inhibitor to a patient at risk of atherosclerosis progression, to suppress the proliferation of vascular smooth muscle cells in said patient, whereby the risk of atherosclerosis progression in the patient is reduced.  
     
     
         14 . The method of  claim 5  wherein said contacting comprises administering said inhibitor to a patient at risk of keloid formation, whereby keloid formation in said patient is inhibited.  
     
     
         15 . The method of  claim 5  wherein said contacting comprises administering said inhibitor to a patient suffering from cancer originating from a smooth muscle cell, whereby proliferation of a cancer cell is inhibited.  
     
     
         16 . A method for identifying an inhibitor of smooth muscle cell proliferation comprising: 
 extracting RNAs from growing vascular smooth muscle cells from a first animal model that is restenosis-resistant with respect to balloon injury to a blood vessel in said first animal model, to provide a first pool of isolated RNAs;    generating a first cDNA pool from said first pool of RNAs;    extracting RNAs from growing vascular smooth muscle cells from a second animal model that is restenosis-prone with respect to balloon injury of a blood vessel in said second animal model, to provide a second pool of isolated RNAs;    generating a second cDNA pool from said second pool of isolated RNAs;    identifying cDNAs that are present in said first cDNA pool in a greater amount than in said second cDNA pool, to provide a pool of upregulated cDNAs;    identifying a first set of genes that correspond to said upregulated cDNAs;    out of said first set of genes, identifying a subset of genes that encode soluble proteins which are secreted to a greater extent by vascular smooth muscle cells from said first animal model than from said second animal model, to provide a group of upregulated genes encoding soluble proteins;    confirming that at least one said encoded soluble protein is expressed to a greater extent in vascular smooth muscle cells from said first animal model than in vascular smooth muscle cells from said second animal model, to identify at least one upregulated soluble protein inhibitor of smooth muscle cell proliferation;    optionally, expressing at least one gene coding for said upregulated soluble protein;    optionally, purifying said at least one upregulated soluble protein, to provide a purified upregulated soluble protein; and    optionally, adding said purified protein to vascular smooth muscle cells in cell culture medium and confirming that said protein is active for retarding proliferation of said vascular smooth muscle cells.    
     
     
         17 . The method of  claim 16  comprising determining that at least one said upregulated gene is expressed at least 1.5-fold more in said first animal model than in said second animal model.  
     
     
         18 . The method of  claim 16  further comprising identifying a biologically active region of said upregulated soluble protein that is capable of directly or indirectly inhibiting the proliferation of smooth muscle cells.  
     
     
         19 . The method of  claim 19  comprising identifying at least one soluble protein capable of being secreted by a proliferating human vascular smooth muscle cell, having at least 24% amino acid identity to at least one of said protein or proteins encoded by said upregulated gene(s), and having activity for inhibiting smooth muscle cell proliferation.  
     
     
         20 . The method of  claim 19  comprising identifying at least one soluble protein capable of being secreted by a proliferating human vascular smooth muscle cell, having at least 40% amino acid identity to at least one of said protein or proteins encoded by said upregulated gene(s), and having activity for inhibiting smooth muscle cell proliferation.