Treatment of metabolic-related disorders using hypothalamic gene transfer of BDNF and compositions therefor
Described herein is a system which uses a gene therapy particle that includes at least one gene, cDNA, fragment or analogue of at least one neurotropin that binds to the trkB receptor or the trkB receptor itself. The gene therapy particle is capable of being delivered to a subject in need thereof for therapy of a metabolic disorder. In a particular aspect, this invention is directed to an inventive method that demonstrates a remarkable efficacy on lowering body weight.
1. A gene therapy particle comprising:
a first expression cassette comprised of a heterologous promoter operatively linked to a nucleotide coding sequence (cDNA) encoding a brain derived neurotrophic factor (BDNF) which is linked to WPRE; and,
at least a second expression cassette comprised of cDNA AGRP484 nt 568-1051 in SEQ ID NO:7) linked to miR-Bdnf which is, in turn, linked to WPRE74 having SEQ ID NO:3.
2. The gene therapy particle of claim 1 , wherein the gene therapy particle further comprises an adeno-associated viral vector, lentiviral vector or adenoviral vector.
3. A gene therapy particle comprising:
a first expression cassette comprised of a heterologous promoter operatively linked to a nucleotide coding sequence (cDNA) encoding a brain derived neurotrophic factor (BDNF) which is linked to WPRE; and
at least a second expression cassette comprised of cDNA AGRP484 (nt 586-1051 in SEQ ID NO: 7) linked to miR-Bdnf which is, in turn, linked to WPRE74 having SEQ ID NO: 3.
4. The gene therapy particle of claim 3 , wherein the adeno-associated viral vector is AAV-2, or a modified form of AAV-2 with an altered tropism.
5. The gene therapy particle of claim 3 , wherein the AAV nucleotide sequences are derived from AAV serotype 1 (AAV-1).
6. The gene therapy particle of claim 1 , wherein the BDNF is a human BDNF.
7. A pharmaceutical composition comprising the gene therapy particle of claim 1 , in a biocompatible pharmaceutical carrier.
8. The gene therapy particle of claim 1 , wherein the heterologous promoter is chicken β-actin (CBA).
9. The gene therapy particle of claim 1 , wherein the promoter of the first expression cassette includes a constitutive promoter.
10. The gene therapy particle of claim 1 , wherein the miR-Bdnf comprises SEQ ID NO: 9.
11. The gene therapy particle of claim 1 , wherein the BDNF is flanked by two loxP sites (flox-BDNF), wherein the BDNF is capable of being subsequently knocked out by a second viral vector delivering Cre recombinase.
12. A nucleic acid molecule comprising: an expression cassette comprising a heterologous promoter operatively linked to a nucleotide coding sequence (cDNA) encoding a brain derived neurotrophic factor (BDNF) linked to WPRE, and a post-transcriptional regulatory element comprised of cDNA AGRP484 (nt 568-1051 in SEQ ID NO: 7) linked to miR-BDNF which is linked to WPRE74 having SEQ ID NO: 3.
13. The nucleic acid molecule of claim 12 , wherein the promoter is chicken β-actin (CBA).
14. The nucleic acid molecule of claim 12 , wherein the BDNF is a human BDNF.
15. The nucleic acid molecule of claim 12 , wherein the BDNF is flanked by two loxP sites (flox-BDNF), wherein the BDNF is capable of being subsequently knocked out by a second viral vector delivering Cre recombinase.
16. A pharmaceutical composition comprising the nucleic acid molecule of claim 12 , in a biocompatible pharmaceutical carrier.
17. The nucleic acid molecule of claim 12 , wherein the miR-Bdnf comprises SEQ ID NO: 9.