Prevention and treatment of vascular disease with recombinant adeno-associated virus vectors encoding apolipoprotein A-I and apolipoprotein A-I milano
Described herein is a gene therapeutic approach to the prevention and treatment of vascular disease and coronary heart disease; in particular, atherosclerosis. The inventive methods may be used in the prevention and treatment of atherosclerosis, as well as any disease or physiological condition in which atherosclerosis plays a role. The inventive methods involve the gene delivery of ApoA-I or ApoA-IMilano. This may be accomplished by the use of rAAV technology. rAAV virions may be delivered to a mammalian subject by various methodologies, including transplantation of transduced bone marrow cells, direct intramuscular injection, intravenous or portal vein injection or stent delivery.
1. A method of treating atherosclerosis in a mammal, comprising:
providing a recombinant adeno-associated viral (rAAV) particle comprising:
an exogenous gene encoding ApoA-I Milano ; and
AAV rescue and packaging components derived from an AAV serotype selected from the group of consisting of AAV1, AAV2, and AAV5 and combinations thereof; and
delivering said rAAV particle in a single dose to said mammal in an amount sufficient to treat atherosclerosis for at least 20 weeks.
2. The method of claim 1 , wherein said rAAV particle is produced by the process of:
(i) providing a first plasmid that comprises said exogenous gene,
(ii) providing a second plasmid that is complementary to the first plasmid and which comprises components for rescue and packaging,
(iii) co-transfecting the first and second plasmids into a host cell, and
(iv) generating a quantity of said rAAV particle from said co-transfected host cell,
wherein the pair of said first and second plasmids is selected such that said rAAV particle is targeted for delivery to a specific tissue type.
3. The method of claim 2 , wherein said second plasmid further comprises AAV rescue and packaging components derived from an AAV serotype selected from the group consisting of AAV1, AAV2, AAV5, and combinations thereof.
4. The method of claim 1 , wherein said mammal is a human.
5. The method of claim 1 , wherein said amount sufficient to treat atherosclerosis is from about 1×10 10 rAAV genome/kg of said mammal to about 1×10 14 rAAV genome/kg of said mammal.
6. The method of claim 1 , wherein said rAAV particle is delivered to said mammal intramuscularly, intravenously, or both.
7. The method of claim 1 , wherein providing said rAAV particle, further comprises transducing multipotent stem cells with a quantity of said rAAV particle, and delivering said rAAV particle further comprises transplanting said multipotent stem cells into said mammal.
8. The method of claim 7 , wherein said cells are bone marrow cells.
9. A kit for the treatment of atherosclerosis in a mammal, comprising:
a volume of recombinant adeno-associated viral (rAAV) particle comprising:
an exogenous gene encoding ApoA-I Milano ;
AAV rescue and packaging components derived from an AAV serotype selected from the group of consisting of AAV1, AAV2, and AAV5 and combinations thereof; and
instructions for the use of said volume of rAAV particle for treating atherosclerosis in said mammal.
10. The method of claim 1 , wherein the recombinant rAAV particle comprises AAV rescue and packaging components derived from AAV2.
11. The kit of claim 9 , wherein the recombinant rAAV particle comprises AAV rescue and packaging components derived from AAV2.