Selective gene therapy expression system
The present invention relates to an expression system for systemic administration comprising a sequence encoding a protein, said expression system allowing: the expression at a therapeutically acceptable level of the protein in the target tissues including skeletal muscles; and the expression at toxically acceptable level of the protein in tissues other than the target tissues, especially in the heart.
1. An expression vector for systemic administration of a sequence encoding calpain 3, comprising:
the sequence encoding calpain 3;
a muscle-specific promoter sequence that regulates expression of the sequence encoding calpain 3 in skeletal muscles, wherein the muscle-specific promoter sequence comprises a promoter sequence of calpain 3, a promoter sequence of miR206, a promoter sequence of skeletal alpha-actin, a promoter sequence of troponin, or a fragment thereof; and
at least one target sequence of miR208a that comprises SEQ ID NO: 10.
2. The expression vector according to claim 1 , wherein the calpain 3 comprises SEQ ID NO: 7.
3. The expression vector according to claim 1 , wherein the promoter sequence of calpain 3 comprises SEQ ID NO: 12.
4. The expression vector according to claim 1 , wherein the promoter sequence of miR206 comprises SEQ ID NO: 13.
5. The expression vector according to claim 1 , wherein the vector is a viral vector.
6. The expression vector according to claim 1 , wherein the vector comprises an adeno-associated viral vector (AAV).
7. The expression vector according to claim 6 , wherein the AAV is AAV8 or AAV9 serotype.
8. A pharmaceutical composition comprising the expression vector according to claim 1 and a pharmaceutically acceptable carrier.
9. The expression vector according to claim 1 , wherein the muscle-specific promoter sequence is a skeletal muscle-specific promoter sequence.
10. The expression vector according to claim 1 , wherein the muscle-specific promoter sequence is a promoter sequence of skeletal alpha-actin or a fragment thereof.
11. The expression vector according to claim 1 , wherein the muscle-specific promoter sequence is a promoter sequence of troponin or a fragment thereof.
12. The pharmaceutical composition of claim 8 , wherein the pharmaceutical composition is formulated for systemic administration.
13. The pharmaceutical composition of claim 8 , wherein the pharmaceutical composition is formulated in liquid form.
14. The expression vector according to claim 1 , wherein the muscle-specific promoter sequence comprises a promoter sequence of skeletal alpha-actin or a fragment thereof.
15. An expression AAV vector for systemic administration of a sequence encoding calpain 3, comprising:
an AAV vector selected from the group consisting of a AAV8 and AAV9 vectors, variants thereof or artificial serotypes, and chimeric AAV vectors;
the sequence encoding calpain 3;
a muscle-specific promoter sequence that regulates expression of the sequence encoding calpain 3 in skeletal muscles, wherein the muscle-specific promoter sequence comprises a promoter sequence of calpain 3, a promoter sequence of skeletal alpha-actin, a promoter sequence of troponin, a promotor sequence of miR206, or a fragment thereof; and
at least one target sequence of miR208a that comprises SEQ ID NO: 10.
16. The expression vector according to claim 15 , wherein the muscle-specific promoter sequence is a promoter sequence of skeletal alpha-actin or a fragment thereof.