Gene-therapy vectors for treating cardiomyopathy
The present invention relates to a gene therapy vector which is useful in the treatment or prevention of hypertrophic cardiomyopathy in a subject in need thereof. The gene therapy vector of the invention comprises a nucleic acid sequence encoding a cardiac sarcomeric protein and a cardiomyocyte-specific promoter which is operably linked to said nucleic acid sequence. The invention furthermore relates to a cell which comprises the gene therapy vector. Pharmaceutical compositions which comprise the gene therapy vector and/or a cell comprising said vector are also provided. In another aspect, the invention relates to a method for treating or preventing hypertrophic cardiomyopathy in a subject by introducing the gene therapy vector of the invention into a subject in need of treatment.
1. A gene therapy vector for expressing an exogenous nucleic acid sequence comprising:
(a) a nucleic acid sequence encoding a functional cardiac myosin binding protein C (cMyBP-C), and
(b) a human cardiac troponin T (hTNNT2) promoter which is operably linked to said nucleic acid sequence,
wherein said gene therapy vector, and wherein the hTNNT2 promoter comprises a nucleic acid sequence at least 95% identical over its length compared to SEQ ID NO: 5 is an adeno-associated virus (AAV) vector.
2. The gene therapy vector of claim 1 , wherein the hTNNT2 promoter comprises the nucleic acid sequence of SEQ ID NO: 5.
3. The gene therapy vector of claim 1 wherein said cMyBP-C protein comprises the cMyBP-C amino acid sequence of SEQ ID NO: 2 or a functional variant thereof having at least 80% sequence identity thereto.
4. The gene therapy vector of claim 1 , wherein said cMyBP-C protein comprises the cMyBP-C amino acid sequence of SEQ ID NO: 2 or a functional variant thereof having at least 90% sequence identity thereto.
5. The gene therapy vector of claim 1 , wherein said cMyBP-C protein comprises the cMyBP-C amino acid sequence of SEQ ID NO: 2.
6. The gene therapy vector of claim 3 , further comprising an intron which increases gene expression levels, said intron comprising a fragment of beta globin gene intron.
7. The gene therapy vector of claim 3 , further comprising intron sequences from the human beta globin gene and human immunoglobulin G (IgG).
8. The gene therapy vector of claim 3 , further comprising an intron comprising the nucleic acid sequence of SEQ ID NO: 7.
9. The gene therapy vector of claim 1 , wherein the AAV vector is an AAV serotype 1 vector, AAV serotype 6 vector, AAV serotype 8 vector, or AAV serotype 9 vector.
10. The gene therapy vector of claim 1 , wherein the nucleic acid sequence encoding the functional cMyBP-C protein and the hTNNT2 promoter are within a polynucleotide insert having a size of at least 4.0 kbp, up to 5.4 kbp.
11. The gene therapy vector of claim 1 , wherein the nucleic acid sequence encoding the functional cMyBP-C protein and the hTNNT2 promoter are within a polynucleotide insert having a size of at least 4.5 kbp, up to 5.4 kbp.
12. The gene therapy vector of claim 3 , wherein the nucleic acid sequence encoding the functional cMyBP-C protein and the hTNNT2 promoter are within a polynucleotide insert having a size of at least 4.0 kbp, up to 5.4 kbp.
13. The gene therapy vector of claim 3 , wherein the nucleic acid sequence encoding the functional cMyBP-C protein and the hTNNT2 promoter are within a polynucleotide insert having a size of at least 4.5 kbp, up to 5.4 kbp.
14. The gene therapy vector of claim 1 comprising
(a) a hTNNT2 promoter that comprises the nucleic acid sequence of SEQ ID NO: 5,
(b) a nucleic acid sequence encoding the cMyBP-C amino acid sequence of SEQ ID NO: 2, and
(c) an intron comprising the nucleic acid sequence of SEQ ID NO: 7.
15. The gene therapy vector of claim 1 comprising:
(a) a hTNNT2 promoter that comprises the nucleic acid sequence of SEQ ID NO: 5, and
(b) a nucleic acid sequence encoding the cMyBP-C amino acid sequence of SEQ ID NO: 2.