Compartmentalized method of nucleic acid delivery and compositions and uses thereof

Abstract

Provided herein is a method of delivering nucleic acid molecules to a compartmentalized tissue or organ of a subject. Also provided herein are uses and processes for delivering a nucleic acid molecule to the parenchyma of a compartmentalized tissue or organ. The methods and uses can be used in the treatment of diseases and conditions and in industrial, agricultural and veterinary applications. Also provided herein are compositions containing an adenovirus or adeno-associated virus or other recombinant virus formulated for administration to the parenchyma of a tissue or organ.

What is claimed: 
     
         1 . A method of delivering a nucleic acid molecule to a subject, comprising:
 a) compartmentalizing a tissue or organ or portion of the tissue or organ from systemic circulation of the subject, whereby the tissue, organ or portion thereof is isolated from systemic circulation;   b) injecting a delivery agent directly to the parenchyma of the compartmentalized tissue or organ or portion of the tissue or organ, wherein the delivery agent comprises the nucleic acid molecule, wherein, upon injection directly into the parenchyma, the delivery agent enters parenchymal cells of the compartmentalized tissue or organ or portion of the tissue or organ, and is not delivered into systemic circulation;   c) maintaining the compartmentalization after administering the delivery agent for a time sufficient to effect quantitative cellular uptake of the delivery agent into cells of the parenchyma; and   d) after the time to effect cellular uptake of the delivery agent, restoring communication between the tissue, organ or portion of the tissue or organ and the systemic circulation.   
     
     
         2 . The method of  claim 1 , wherein compartmentalization is effected by clamping the tissue, organ or portion thereof. 
     
     
         3 . The method of  claim 1 , wherein the time sufficient to effect quantitative cellular uptake is more than 15 minutes. 
     
     
         4 . The method of  claim 3 , wherein the time sufficient to effect quantitative cellular uptake is up to 60 minutes. 
     
     
         5 . The method of  claim 1  wherein the tissue or organ or portion thereof is the liver or a portion thereof. 
     
     
         6 . The method of  claim 5 , wherein the portion has a volume of at least 5 mm 3 . 
     
     
         7 . The method of  claim 1 , wherein compartmentalization is achieved by applying pressure to the organ, tissue or portion thereof with a clamp, thereby compressing any arteries, veins, ducts and/or vessels and blocking the blood supply and flow to the arteries, veins, ducts and/or vessels in the tissue or organ or portion of the tissue or organ that is clamped, thereby eliminating the blood supply and flow to the tissue or organ or portion thereof that is clamped. 
     
     
         8 . The method of  claim 5 , wherein a portion of the liver is compartmentalized and the portion is a lobe, segment or a portion of a lobe or segment of the liver. 
     
     
         9 . The method of  claim 8 , wherein the lobe or portion of a lobe is selected from among the right lobe, the left lobe, the quadrate lobe and the caudate lobe or is a portion thereof. 
     
     
         10 . The method of  claim 1 , wherein the delivery agent is selected from among a non-viral vector, a virus, a virus-like particle, a minicircle, a nanoparticle and a whole cell that comprise the nucleic acid molecule. 
     
     
         11 . The method of  claim 10 , wherein the delivery agent is a virus and the virus is selected from among an adenovirus, an adeno-associated virus (AAV), a retrovirus, vaccinia virus and herpes simplex virus. 
     
     
         12 . The method of  claim 1 , wherein the delivery agent is a virus or viral vector that comprises a heterologous nucleic acid encoding a therapeutic protein. 
     
     
         13 . The method of  claim 12 , wherein the virus is a lentivirus. 
     
     
         14 . The method of  claim 12 , wherein the virus is an adenovirus. 
     
     
         15 . The method of  claim 14 , wherein the adenovirus is adenovirus type 2 or adenovirus type 5. 
     
     
         16 . The method of  claim 15 , wherein the encoded polypeptide is selected from among an enzyme, a hormone, a coagulation or clotting factor, a cytokine, a growth factor or active portion thereof, an antibody or antigen binding portions of antibodies, an angiogenesis modulator, an immunomodulator, a pain modulator, a receptor or active portion thereof, a transport protein, a regulatory protein, an antigen and an allergen. 
     
     
         17 . The method of  claim 1 , wherein the nucleic acid encodes insulin. 
     
     
         18 . The method of  claim 1 , wherein the nucleic acid encodes a coagulation or clotting factor. 
     
     
         19 . A composition, comprising a therapeutic nucleic acid molecule, wherein:
 the composition contains a delivery agent comprising the therapeutic nucleic acid molecule;   the delivery agent is a virus or viral vector that comprises the therapeutic nucleic acid molecule;   the nucleic acid molecule encodes a polypeptide;   the composition is formulated as a single dose for administration without dilution into the parenchyma of a tissue, organ, or portion of a tissue or organ that is to be compartmentalized for at least 25 minutes up to 1 hour following administration for uptake of the administered virus or viral vector;   a single dose for the composition contains 1×10 3  to 1×10 12  viral particles or vectors or plaque forming units (pfu) of virus to effect gene therapy; and   a compartmentalized tissue or organ or portion of a tissue or organ is a tissue or organ that has been isolated from the systemic circulation by clamping the tissue, organ, or portion thereof.   
     
     
         20 . The composition of  claim 19 , wherein the delivery agent is a virus or viral vector that encodes the polypeptide. 
     
     
         21 . The composition of  claim 19 , wherein the tissue, organ or portion thereof, is the liver or a portion of the liver.