Gene transfer into the kidney

Abstract

This invention pertains to a method of infecting an organ or a tissue other than a liver with an effective amount of a vector carrying genetic material of interest. This invention features a method of infecting kidney cells by introducing into the vasculature of a kidney a vector carrying genetic material of interest and maintaining the vector in contact with the renal vasculature for a period of time sufficient to allow infection of kidney cells with an effective amount of the vector, and under conditions which protect the kidney from ischemic damage. This method allows for infection of a significant number of renal endothelial cells. The method of the invention can be used for both in vivo and ex vivo applications.

What is claimed is:  
     
         1 . A method of infecting an organ or a tissue other than a liver, comprising: introducing a vector carrying genetic material of interest into a vasculature of said organ or tissue for a period of time sufficient to allow for infection of said organ or tissue with an effective amount of said vector under conditions which protect said organ or tissue from ischemic damage.  
     
     
         2 . The method of  claim 1 , wherein said organ or tissue is selected from the group consisting of a heart, limb, blood vessels, lung and kidney.  
     
     
         3 . A method of infecting kidney cells, comprising: introducing a vector carrying genetic material of interest into a vasculature of a kidney for a period of time sufficient to allow for infection of said kidney cells with an effective amount of said vector under conditions which protect said kidney from ischemic damage.  
     
     
         4 . The method of  claim 3 , wherein said kidney is protected from ischemic damage by reducing kidney temperature.  
     
     
         5 . The method of  claim 4 , wherein said kidney temperature is reduced by incubating said kidney on ice.  
     
     
         6 . The method of  claim 3 , wherein said vector is introduced in combination with an oxygen supplying agent.  
     
     
         7 . The method of  claim 3 , wherein said vector is introduced in combination with a vasodilator.  
     
     
         8 . The method of  claim 3 , wherein said kidney is a diseased kidney.  
     
     
         9 . The method of  claim 8 , wherein said diseased kidney is afflicted with polycystic kidney disease (PKD).  
     
     
         10 . A method of infecting kidney cells with genetic material of interest comprising: 
 (i) restricting blood flow to and from a kidney;    (ii) introducing into a vasculature of said kidney a vector carrying said genetic material of interest; and    (iii) maintaining said vector in contact with the vasculature of said kidney long enough to allow for infection of said kidney cells with said vector while reducing kidney temperature to a temperature such that said kidney is protected from ischemic damage.    
     
     
         11 . The method of  claim 10 , wherein said vector is introduced intra-arterially or intravenously.  
     
     
         12 . The method of  claim 10 , wherein said blood flow to a kidney is restricted by clamping the aorta.  
     
     
         13 . The method of  claim 12 , wherein said aorta is clamped under the superior mesenteric artery.  
     
     
         14 . The method of  claim 10 , wherein said blood flow from a kidney is restricted by clamping the renal vein.  
     
     
         15 . The method of  claim 10 , wherein said vector is a virus capable of infecting non-dividing cells.  
     
     
         16 . The method of  claim 15 , wherein said virus is selected from the group consisting of adeno associated-virus (AAV), adenovirus and herpesviruses.  
     
     
         17 . The method of  claim 16 , wherein said adenovirus is a replication deficient adenovirus.  
     
     
         18 . The method of  claim 17 , wherein said replication deficient adenovirus is selected from the group consisting of Ad-2, Ad-3, Ad-5 and Ad-7 strain of adenoviruses.  
     
     
         19 . The method of  claim 17 , wherein said replication deficient adenovirus lacks one or more of E1A, E1B or E3.  
     
     
         20 . The method of  claim 16 , wherein said replication deficient adenovirus is introduced in the amount ranging from about 10 10  pfu/ml to about 10 11  pfu/ml.  
     
     
         21 . The method of  claim 10 , wherein said genetic material of interest is selected from the group consisting of cytokines, growth factors, chimeric toxins, adhesion molecules and cell surface molecules.  
     
     
         22 . The method of  claim 10 , wherein said kidney cells of interest are endothelial cells of kidney cortex or medulla.  
     
     
         23 . The method of  claim 10 , wherein the temperature of said kidney is reduced to at least about 0° C. to at least about 25° C.  
     
     
         24 . The method of  claim 10 , wherein the temperature of said kidney is reduced to about 4° C.  
     
     
         25 . The method of  claim 10 , wherein the temperature of said kidney is reduced by contacting the kidney with a cold reagent.  
     
     
         26 . The method of  claim 25 , wherein said cold reagent is ice.  
     
     
         27 . The method of  claim 10 , wherein said kidney is incubated at a reduced temperature for at least about 30 min. to at least about 1 hour.  
     
     
         28 . The method of  claim 10 , wherein said kidney is incubated at a reduced temperature for about 45 minutes.  
     
     
         29 . The method of  claim 10 , wherein said vector is introduced in combination with a vasodilator.  
     
     
         30 . The method of  claim 29 , wherein said vasodilator is selected from the group consisting of dopamine and papaverine.  
     
     
         31 . The method of  claim 10 , wherein said vector is introduced in combination with an oxygen supplying agent.  
     
     
         32 . A method of infecting kidney cells with genetic material of interest comprising: 
 (i) restricting blood flow to and from a kidney;    (ii) introducing into a vasculature of said kidney a vector carrying said genetic material of interest; and    (iii) incubating said kidney on ice for substantially the period of time that said vector is maintained in contact with said vasculature.    
     
     
         33 . A method of infecting kidney cells in situ comprising: 
 (i) restricting blood flow to a kidney by clamping the aorta;    (ii) flushing said kidney with cold saline;    (iii) introducing into said kidney via the renal artery an adenovirus carrying genetic material of interest;    (iv) restricting blood flow from said kidney by clamping the renal vein such that said vector is maintained in contact with the vasculature of said kidney;    (v) reducing kidney temperature to a temperature such that said kidney is protected from ischemic damage long enough to allow for infection of said kidney cells with said vector; and    (vi) restoring blood flow to said kidney.    
     
     
         34 . The method of  claim 33 , wherein said kidney cells are endothelial cells of kidney cortex or medulla.  
     
     
         35 . The method of  claim 33 , wherein said genetic material of interest is selected from the group consisting of cytokines, growth factors, chimeric toxins, adhesion molecules and cell surface molecules.  
     
     
         36 . The method of  claim 33 , wherein said vector is administered in combination with a vasodilator.  
     
     
         37 . The method of  claim 33 , wherein said vasodilator is selected from the group consisting of dopamine and papaverine.  
     
     
         38 . The method of  claim 33 , wherein said vector is introduced in combination with an oxygen supplying agent.  
     
     
         39 . The method of  claim 33 , wherein said kidney temperature is reduced by contacting the kidney with a cold reagent.  
     
     
         40 . The method of  claim 39 , wherein said cold reagent is ice.  
     
     
         41 . The method of  claim 33 , wherein the temperature of said kidney is reduced to at least about 0° C. to at least about 25° C.  
     
     
         42 . The method of  claim 33 , wherein the temperature of said kidney is reduced to about 4° C.  
     
     
         43 . The method of  claim 33 , wherein said kidney is incubated at the reduced temperature for at least about 30 min to at least about 1 hour.  
     
     
         44 . The method of  claim 33 , wherein said kidney is incubated at a reduced temperature for about 45 min.  
     
     
         45 . The method of  claim 33 , wherein said kidney is a diseased kidney.  
     
     
         46 . The method of  claim 45 , wherein said diseased kidney is afflicted with polycystic kidney disease (PKD).  
     
     
         47 . A kidney having endothelial cells infected with a recombinant virus carrying genetic material of interest.  
     
     
         48 . The kidney of  claim 47 , wherein at least about 1% to at least about 15% of said endothelial cells are infected with said recombinant virus.  
     
     
         49 . The kidney of  claim 47 , wherein at least about 10% of said endothelial cells are infected with said recombinant virus.  
     
     
         50 . The kidney of  claim 47 , wherein said recombinant virus is capable of infecting non-dividing cells.  
     
     
         51 . The kidney of  claim 50 , wherein said virus is selected from the group consisting of adeno-associated viruses (AAV), adenoviruses and herpesviruses.  
     
     
         52 . The kidney of  claim 51 , wherein said adenovirus is replication deficient adenovirus.  
     
     
         53 . The kidney of  claim 47 , wherein said genetic material of interest is selected from the group consisting of cytokines, growth factors, chimeric toxins, adhesion molecules and cell surface molecules.