Use of pro-apoptotic factors in treatment of atherosclerosis

Abstract

The present invention features a novel method of treating vascular disease that involves modifying smooth muscle cells to express a gene encoding a protein having both anti-inflammatory and pro-apoptotic activity. Preferably, the protein of the invention also has an anti-proliferative effect in smooth muscle cells. In general, the method is useful in preparing vascularized organs and vessels for transplant into a patient. Alternatively, the present invention can be applied to treat atherosclerotic lesions in damaged vessels.

What is claimed is:  
     
         1 . A method of treating or preventing vascular disease comprising modifying a mammalian smooth muscle cell by inserting into said smooth muscle cell, or a progenitor thereof, DNA encoding a protein capable of decreasing inflammation and increasing apoptosis of said smooth muscle cell and transplanting said smooth muscle cell, or a tissue or organ comprising such a cell, into a transplant candidate or a patient diagnosed with a vascular disease, wherein said protein has A20 activity.  
     
     
         2 . The method of    claim 1   , wherein said vascular disease comprises atherosclerosis, transplant-associated vasculopathy, or chronic rejection.  
     
     
         3 . A method of treating or preventing transplant-associated vasculopathy (TAV) comprising modifying a mammalian smooth muscle cell by inserting into said smooth muscle cell, or a progenitor thereof, DNA encoding a protein capable of decreasing inflammation and increasing apoptosis of said smooth muscle cell and transplanting said smooth muscle cell, or a tissue or organ comprising such a cell into a patient at risk for developing TAV, wherein said protein has A20 activity.  
     
     
         4 . The method of    claim 1    or    3   , wherein said protein is A20.  
     
     
         5 . The method of    claim 1    or    3   , wherein said protein further blocks smooth muscle cell proliferation.  
     
     
         6 . The method of    claim 4   , wherein said A20 comprises a mutant or derivative of said A20.  
     
     
         7 . A method of treating atherosclerosis comprising transplanting donor smooth muscle cells, or graftable tissues or organs comprising smooth muscle cells, to a mammalian recipient having atherosclerosis or a vascularized organ transplant, comprising the steps of (a) modifying the donor smooth muscle cells, or progenitor smooth muscle cells thereof, by inserting therein DNA encoding A20; and (b) transplanting the resultant modified donor smooth muscle cells, or tissues or organ comprising the cells into the recipient, and expressing in the cells A20, thereby substantially promoting apoptosis.  
     
     
         8 . A method of preventing TAV comprising, transplanting donor smooth muscle cells, or graftable tissues or organs comprising smooth muscle cells, to a mammalian recipient comprising the steps of (a) modifying the donor smooth muscle cells, or progenitor smooth muscle cells thereof, by inserting therein DNA encoding A20; and (b) transplanting the resultant modified donor smooth muscle cells, or tissues or organ comprising the cells into the recipient, and expressing in the cells A20, thereby substantially promoting apoptosis in the presence of a cellular activating stimulus.  
     
     
         9 . The method of    claim 7    or    8   , wherein said method substantially inhibits inflammation in the presence of a cellular activating stimulus.  
     
     
         10 . The method of    claim 7    or    8   , wherein said A20 comprises a mutant or derivatives of A20.  
     
     
         11 . A method of preparing a vascularized organ or vessel for transplantation into a patient comprising obtaining an organ or vessel for transplant and perfusing said organ or vessel with nucleic acid encoding A20 protein.  
     
     
         12 . The method of    claim 11   , wherein said nucleic acid encoding A20 is inserted within a vector comprising an adenovirus vector or a lentivirus vector.  
     
     
         13 . The method of    claim 11   , further comprising perfusing said organ or vessel with an immunoregulatory factor.  
     
     
         14 . The method of    claim 13   , wherein said immunoregulatory factor is a cytokine.  
     
     
         15 . The method of    claim 13   , wherein said immunoregulatory factor is Fas.  
     
     
         16 . The method of    claim 13   , wherein said perfusing occurs in vivo.  
     
     
         17 . The method of    claim 13   , wherein said perfusing occurs ex vivo.  
     
     
         18 . The method of    claim 12   , wherein said A20 comprises a mutant or derivative of said A20.  
     
     
         19 . The method of    claim 11   , further comprising enhancing the expression of A20 using a therapeutic agent.  
     
     
         20 . The method of    claim 11   , wherein said patient has suffered ischemia, reperfusion injury, mechanical injury, immunologic injury, pharmacologic injury, or coronary trauma.  
     
     
         21 . The method of    claim 11   , wherein said patient has undergone balloon angioplasty.  
     
     
         22 . The method of    claim 11   , wherein said patient has atherosclerosis.  
     
     
         23 . The method of    claim 11   , wherein said organ or vessel is being prepared as a vascularized organ or a vascular graft.  
     
     
         24 . A mammalian smooth muscle cell which is modified to express A20, said A20 being capable of inhibiting inflammation and promoting apoptosis in the presence of a cellular activating stimulus, said modified smooth muscle cell exhibiting increased sensitivity to apoptosis, relative to an untreated smooth muscle cell.  
     
     
         25 . The smooth muscle cell of    claim 24   , wherein said A20 further blocks smooth muscle cell proliferation.  
     
     
         26 . The smooth muscle cell of    claim 24   , wherein said A20 comprises a mutant or derivative of A20.  
     
     
         27 . A donor smooth muscle cell, or a tissue or organ comprising such a cell, for transplantation into a recipient species, wherein said cell is modified to express A20, said A20 being capable of inhibiting inflammation and promoting apoptosis in the presence of a cellular activating stimulus in said recipient.  
     
     
         28 . The smooth muscle cell of    claim 27   , wherein said A20 further blocks smooth muscle cell proliferation.  
     
     
         29 . The smooth muscle cell of    claim 27   , wherein said A20 comprises a mutant or derivative of A20.  
     
     
         30 . An organ or vessel from a non-human transgenic or somatic recombinant mammal comprising DNA encoding a pro-apoptotic protein of a different species, wherein said protein has A20 activity.  
     
     
         31 . An organ or vessel according to    claim 30    which is porcine.  
     
     
         32 . An organ or vessel according to    claim 30   , wherein said pro-apoptotic protein is human.  
     
     
         33 . A method of identifying compounds that substantially promote apoptosis and substantially inhibit growth or proliferation in smooth muscle cells comprising, 
 a) exposing smooth muscle cells to a compound; and    b) measuring activation of A20 in smooth muscle cells.    
     
     
         34 . A method of identifying compounds that substantially promote apoptosis and substantially inhibit growth or proliferation in smooth muscle cells comprising, 
 a) exposing smooth muscle cells to a compound; and    b) measuring binding of said compound to a pro-apoptotic protein, wherein said protein is A20.    
     
     
         35 . A method of treating a patient having a vascularized organ transplant, a vessel transplant, or vascular disease comprising enhancing A20 biological activity.  
     
     
         36 . A method of preventing vascular disease in a patient comprising enhancing A20 biological activity.  
     
     
         37 . The method of    claim 35    or    36   , wherein said method comprises administering to said patient a therapeutic agent.  
     
     
         38 . The method of    claim 37   , wherein said therapeutic agent comprises a cytokine.  
     
     
         39 . The method of    claim 37   , wherein said therapeutic agent comprises Fas.  
     
     
         40 . The method of    claim 37   , wherein said therapeutic agent comprises a drug.  
     
     
         41 . A method of promoting apoptosis in smooth muscle cells in a patient, said method comprising increasing the level of A20 in said smooth muscle cells.  
     
     
         42 . The method of    claim 41   , wherein said smooth muscle cell is engineered to express A20.  
     
     
         43 . The method of    claim 36    or    42   , wherein said A20 protein comprises a mutant or derivative of said A20 protein.  
     
     
         44 . The method of    claim 41   , wherein said method comprises administering to said patient a therapeutic agent.  
     
     
         45 . The method of    claim 44   , wherein said therapeutic agent comprises a transcription factor.  
     
     
         46 . The method of    claim 44   , wherein said therapeutic agent comprises a cytokine.  
     
     
         47 . The method of    claim 44   , wherein said therapeutic agent comprises a drug.  
     
     
         48 . The method of    claim 44   , wherein said therapeutic agent comprises Fas.  
     
     
         49 . The method of    claim 41   , wherein said patient has a vascularized organ transplant, a vessel transplant, or vascular disease.  
     
     
         50 . The method of    claim 49   , wherein said vascularized organ comprises the heart, liver, or kidney.  
     
     
         51 . The method of    claim 49   , wherein said vascular disease comprises atherosclerosis or TAV.  
     
     
         52 . A method of inducing cell death of undesired proliferating smooth muscle cells within the neointima of a diseased vessel, said method comprising, enhancing A20 biological activity in said smooth muscle cell.  
     
     
         53 . A method of inducing cell death of undesired proliferating smooth muscle cells within the neointima of a diseased vessel, said method comprising, increasing the level of A20 in said smooth muscle cell.  
     
     
         54 . The method of    claim 52    or    53   , wherein said method comprises administering to said smooth muscle cell a therapeutic agent.  
     
     
         55 . A method of treating or preventing vascular disease in a patient comprising, preparing a smooth muscle cell, or tissue or organ comprising a smooth muscle cell, for transplant by treating said smooth muscle cell, or tissue or organ comprising a smooth muscle cell, with a therapeutic agent that increases A20 biological activity in said smooth muscle cell, or tissue or organ comprising said smooth muscle cell, and transplanting said smooth muscle cell, or tissue or organ comprising said smooth muscle cell, treated with said therapeutic agent to a patient at risk for developing vascular disease.  
     
     
         56 . A method of treating or preventing vascular disease in a patient comprising preparing a smooth muscle cell, or tissue or an organ comprising a smooth muscle cell, for transplant by treating said smooth muscle cell, or tissue, or organ comprising a smooth muscle cell, with a therapeutic agent that increases the A20 level in said smooth muscle cell, or tissue or organ comprising said smooth muscle cell, and transplanting said smooth muscle cell, or tissue or organ comprising said smooth muscle cell, treated with said therapeutic agent to a patient at risk for developing vascular disease.