Hepatocyte Based Insulin Gene Therapy For Diabetes
A method and vectors for controlling blood glucose levels in a mammal are disclosed. In one embodiment, the method comprises the steps of: treating the hepatocyte cells of a patient with a first, second or third vector, wherein the first vector comprises a promoter enhancer, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase and an albumin 3′UTR and lacks an HGH intron, wherein the second vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase site and an albumin 3′UTR and lacks a promoter enhancer, wherein the third vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase site, an albumin 3′UTR and a promoter enhancer and observing the patient's insulin levels, wherein the patient's insulin levels are controlled.
1 . A method for obtaining glucose-regulated expression of insulin ex vivo in hepatocyte cells, wherein the method comprises delivering a first, second or third genetic vector for glucose-regulated synthesis of insulin into an isolated hepatocyte cell,
wherein the first vector comprises a promoter enhancer, 1-5 glucose inducible regulatory elements, a liver-specific promoter, a gene encoding insulin with modified peptidase sites and an albumin 3′UTR and lacks an HGH intron,
wherein the second vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding insulin with modified peptidase sites and an albumin 3′UTR and lacks a promoter enhancer,
wherein the third vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding insulin with modified peptidase sites, an albumin 3′UTR and a promoter enhancer, and
wherein glucose-regulated expression of insulin occurs.
2 . The method of claim 1 additionally comprising the step of transplanting the hepatocytes back into a mammal.
3 . The method of claim 1 , wherein the genetic vector is delivered by exposing the cells to a virus infective for the cells, wherein the virus comprises the genetic construct, and whereby at least a portion of the cells are infected by the virus under suitable conditions and at a sufficient multiplicity.
4 . The method of claim 1 wherein the mammal is human.
5 . The method of claim 1 wherein the insulin is human insulin.
6 . A vector suitable for controlling blood glucose levels in a mammal,
wherein the vector comprises a promoter enhancer, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding insulin with modified peptidase sites and an albumin 3′UTR and lacks an HGH intron or
wherein the vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding insulin with modified peptidase sites and an albumin 3′UTR and lacks a promoter enhancer or
wherein the vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase sites, an albumin 3′UTR and a promoter enhancer.
7 . The vector of claim 6 wherein additionally comprising a VEGF-Enhancer.
8 . The vector of claim 6 wherein the promoter is an albumin promoter.
9 . The vector of claim 6 wherein the insulin is human insulin.
10 . A method of controlling blood glucose levels in a mammal, comprising the steps of:
treating a mammal with a first, second or third vector,
wherein the first vector comprises a promoter enhancer, 1-5 glucose inducible regulatory elements, a liver-specific promoter, a gene encoding insulin with modified peptidase sites and an albumin 3′UTR and lacks an HGH intron and
wherein the second vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding insulin with modified peptidase sites and an albumin 3′UTR and lacks a promoter enhancer,
wherein the third vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding insulin with modified peptidase sites and an albumin 3′UTR and a promoter enhancer, and
observing the mammal's insulin levels, wherein the mammal's insulin levels are controlled.
11 . The method of claim 1 wherein the vector is in a minicircle format.
12 . The method of claim 10 wherein the vector is in a minicircle format.
13 . The method of claim 10 wherein the mammal is human.
14 . The method of claim 13 wherein the insulin is human insulin.
15 . The method of claim 10 wherein the mammal's cholesterol level decreases after treatment.
16 . The method of claim 10 wherein the mammal's triglyceride level decreases after treatment.
17 . The method of claim 10 wherein the mammal is a cat.
18 . The method of claim 10 wherein the mammal is a dog.
19 . The method of claim 10 wherein the mammal is selected for the group consisting of hamsters, gerbils, rats, mice, rabbits, guinea pigs, chinchillas and ferrets.
20 . The method of claim 10 wherein the mammal is a non-human mammal.
21 . The method of claim 10 wherein the patient has a decrease in the plasma level of a compound selected from the group of AST, ALT, and alkaline phosphatase after treatment.