Methods for the treatment of Danon disease and other disorders of autophagy
This disclosure provides gene therapy vectors comprising a polynucleotide encoding one or more isoforms of lysosome-associated membrane protein 2 (LAMP-2), and methods of using such gene therapy vectors for the treatment of Danon disease and other autophagy disorders.
1. A serotype 9 adeno-associated virus (AAV9) vector comprising a polynucleotide encoding lysosome-associated membrane protein 2B (LAMP-2B) operatively linked to a hybrid promoter comprising a chicken beta-actin promoter and a CMV enhancer (CAG promoter).
2. The vector of claim 1 , wherein the expression cassette comprises operably linked in the 5′ to 3′ direction, a first inverse terminal repeat, the CAG promoter, the polynucleotide encoding LAMP-2B, a 3′ untranslated region including a polyadenylation signal, and a second inverse terminal repeat.
3. The vector of claim 1 , wherein the LAMP-2B is human LAMP-2B.
4. The vector of claim 1 , wherein the polynucleotide encoding LAMP-2B has at least about 90% sequence identity to the nucleic acid sequence of SEQ ID NO:2.
5. The gene therapy vector of claim 4 , wherein the polynucleotide encoding LAMP-2B comprises the nucleic acid sequence of SEQ ID NO:2.
6. A method of treating impaired autophagic flux in a mammal that has Danon disease, the method comprising:
intravenously administering an adeno-associated virus 9 (AAV9) vector to the mammal that has impaired autophagic flux, a mutation in a lysosomal-associated protein 2 (LAMP2) gene, and Danon disease;
wherein the AAV9 vector comprises a polynucleotide encoding lysosome-associated membrane protein 2B (LAMP-2B) operatively linked to a hybrid promoter comprising a chicken beta-actin promoter and a CMV enhancer (CAG promoter) such that
the impaired autophagic flux is improved.
7. The method of claim 6 , wherein the vector is administered to the mammal multiple times.
8. The method of claim 6 , wherein the mammal has reduced or non-detectable LAMP-2B expression.
9. The method of claim 6 , wherein the AAV9 vector comprises operably linked in the 5′ to 3′ direction, a first inverse terminal repeat, the CAG promoter, the polynucleotide encoding LAMP-2B, a 3′ untranslated region including a polyadenylation signal, and a second inverse terminal repeat.
10. The method of claim 6 , wherein the LAMP-2B is human LAMP-2B.
11. The method of claim 6 , wherein the polynucleotide encoding LAMP-2B has at least about 90% sequence identity to the nucleic acid sequence of SEQ ID NO:2.
12. The method of claim 6 , wherein the polynucleotide encoding LAMP-2B comprises the nucleic acid sequence of SEQ ID NO:2.
13. The method of claim 12 , wherein the vector is administered to the mammal multiple times.
14. The method of claim 12 , wherein the mammal has reduced or non-detectable LAMP-2B expression.