Cardiac-specific protein targeting domain
View Patent ↗The present invention relates to Cardiac Targeting Peptides or CTPs that are able to transduce cardiomyocytes specifically in culture and in vivo, and to methods for using such peptides and their derivatives to deliver peptides, proteins or nucleic acids specifically to the heart. It is based, at least in part, on the discovery that the peptide APWHLSSQYSRT (SEQ ID NO:1) functioned as a cardiac-specific protein targeting peptide and was successful in delivering a number of different cargoes to cardiac muscle cells in vitro and in vivo.
1. A method of introducing a cargo into a cardiac muscle cell comprising administering, to the cardiac muscle cell, an amount of a complex comprising a cardiac targeting peptide linked to a cargo effective to introduce the cargo into the muscle cell, wherein the cardiac targeting peptide comprises the sequence APWHLSSQYSRT (SEQ ID NO:1).
2. The method of claim 1 , wherein the cargo comprises a radioisotope, fluorescent marker, gadolinium marker, luciferase marker, microsphere or nanoparticle.
3. A method of introducing a cargo into a cardiac muscle cell comprising administering, to the cardiac muscle cell, an amount of a complex comprising a cardiac targeting peptide linked to a cargo effective to introduce the cargo into the muscle cell,
wherein the cardiac targeting peptide
(a) is between 8 and 12 amino acids in length and comprises the sequence HLSSQYSR (SEQ ID NO:2), with the proviso that when the cardiac targeting peptide is 12 amino acids in length it is APWHLSSQX 1 SRT (SEQ ID NO:7) where X 1 is W; or
(b) is between 8 and 12 amino acids in length and comprises the sequence HLSSQYSR (SEQ ID NO:2) in which one amino acid of HLSSQYSR (SEQ ID NO:2) is either deleted or substituted by another amino acid.
4. The method of claim 3 , wherein the cardiac targeting peptide is selected from the group consisting of:
APWHLSSQYSR;
(SEQ ID NO: 3)
PWHLSSQYSRT;
(SEQ ID NO: 4)
PWHLSSQYSR;
(SEQ ID NO: 5)
APWHLSSQ X 1 SRT
(SEQ ID NO: 7)
where X 1 is W;
PX 1 HLSSQYSRT
(SEQ ID NO: 8)
where X 1 is W or Y;
PWHLSSQ X 1 SRT
(SEQ ID NO: 9)
where X 1 is W or Y;
X 1 HLSSQYSRT
(SEQ ID NO: 10)
where X 1 is W or Y;
WHLSSQ X 1 SRT
(SEQ ID NO: 11)
where X 1 is W or Y;
X 1 HLSSQYSR
(SEQ ID NO: 12)
where X 1 is W or Y;
and
WHLSSQ X 1 SR
(SEQ ID NO: 13)
where X 1 is W or Y.
5. The method of claim 3 , wherein the cargo comprises a radioisotope, fluorescent marker, gadolinium marker, luciferase marker, microsphere or nanoparticle.
6. A method of treating a human subject suffering from a myocardial infarction, comprising introducing a cargo into a cardiac muscle cell of the human subject comprising administering, to the human subject, a therapeutically effective amount of a complex comprising a cardiac targeting peptide linked to a cargo, where the cargo inhibits cell death, inhibits arrhythmias, improves contractility, lengthens subject survival, or a combination thereof,
wherein the cardiac targeting peptide
(a) is between 8 and 12 amino acids in length and comprises the sequence HLSSQYSR (SEQ ID NO:2) , with the proviso that when the cardiac targeting peptide is 12 amino acids in length it is selected from the group consisting of APX 1 HLSSQYSRT (SEQ ID NO:6) where X 1 is Y, and APWHLSSQX 1 SRT (SEQ ID NO:7) where X 1 is W; or
(b) is between 8 and 12 amino acids in length and comprises the sequence HLSSQYSR (SEQ ID NO:2) in which one amino acid of HLSSQYSR (SEQ ID NO:2) is either deleted or substituted by another amino acid.
7. The method of claim 6 where the cargo is selected from an NE-κB inhibitor, NSD peptide, hemeoxygenase, an antioxidant, iNOS, S100A1, superoxide dismutase, catalase, glutathione peroxidase, a TGFβ inhibitor, VEGF, FGF-1, FGF-2, sonic hedgehog protein, HGF and an IAP.
8. The method of claim 6 , wherein the cardiac targeting peptide is selected from the group consisting of:
APWHLSSQYSR;
(SEQ ID NO: 3)
PWHLSSQYSRT;
(SEQ ID NO: 4)
PWHLSSQYSR;
(SEQ ID NO: 5)
APWHLSSQ X 1 SRT
(SEQ ID NO: 7)
where X 1 is W;
PX 1 HLSSQYSRT
(SEQ ID NO: 8)
where X 1 is W or Y;
PWHLSSQ X 1 SRT
(SEQ ID NO: 9)
where X 1 is W or Y;
X 1 HLSSQYSRT
(SEQ ID NO: 10)
where X 1 is W or Y;
WHLSSQ X 1 SRT
(SEQ ID NO: 11)
where X 1 is W or Y;
X 1 HLSSQYSR
(SEQ ID NO: 12)
where X 1 is W or Y;
and
WHLSSQ X 1 SR
(SEQ ID NO: 13)
where X 1 is W or Y.
9. A method of treating a human subject suffering from a myocardial infarction, comprising introducing a cargo into a cardiac muscle cell of the human subject comprising administering, to the human subject, a therapeutically effective amount of a complex comprising a cardiac targeting peptide linked to a cargo, where the cargo inhibits cell death, inhibits arrhythmias, improves contractility, lengthens subject survival, or a combination thereof, where the cardiac targeting peptide has the sequence APWHLSSQYSRT (SEQ ID NO:1).
10. The method of claim 9 where the cargo is selected from an NF-κB inhibitor, NBD peptide, hemeoxygenase, an antioxidant, iNOS, S100A1, superoxide dismutase, catalase, glutathione peroxidase, a TGFβ inhibitor, VEGF, FGF-1, FGF-2, sonic hedgehog protein, HGF and an IAP.
11. A method of treating a subject suffering from a metabolic defect that damages the heart comprising introducing a cargo into a cardiac muscle cell of the human subject comprising administering, to the subject, a therapeutically effective amount of a complex comprising a cardiac targeting peptide linked to a cargo, where the cargo corrects the metabolic defect,
wherein the cardiac targeting peptide
(a) is between 8 and 12 amino acids in length and comprises the sequence HLSSQYSR (SEQ ID NO:2), with the proviso that when the cardiac targeting peptide is 12 amino acids in length it is selected from the group consisting of APX 1 HLSSQYSRT (SEQ ID NO:6) where X 1 is Y, and APWHLSSQX 1 SRT (SEQ ID NO:7) where X 1 is W; or
(b) is between 8 and 12 amino acids in length and comprises the sequence HLSSQYSR (SEQ ID NO:2) in which one amino acid of HLSSQYSR (SEQ ID NO:2) is either deleted or substituted by another amino acid.
12. The method of claim 11 , where the metabolic defect is Gaucher's disease and the cargo is glucocerebrosidase.
13. The method of claim 11 , wherein the cardiac targeting peptide is selected from the group consisting of:
APWHLSSQYSR;
(SEQ ID NO: 3)
PWHLSSQYSRT;
(SEQ ID NO: 4)
PWHLSSQYSR;
(SEQ ID NO: 5)
APWHLSSQ X 1 SRT
(SEQ ID NO: 7)
where X 1 is W;
PX 1 HLSSQYSRT
(SEQ ID NO: 8)
where X 1 is W or Y;
PWHLSSQ X 1 SRT
(SEQ ID NO: 9)
where X 1 is W or Y;
X 1 HLSSQYSRT
(SEQ ID NO: 10)
where X 1 is W or Y;
WHLSSQ X 1 SRT
(SEQ ID NO: 11)
where X 1 is W or Y;
X 1 HLSSQYSR
(SEQ ID NO: 12)
where X 1 is W or Y;
and
WHLSSQ X 1 SR
(SEQ ID NO: 13)
where X 1 is W or Y.
14. A method of treating a subject suffering from a metabolic defect that damages the heart comprising introducing a cargo into a cardiac muscle cell of the human subject comprising administering, to the subject, a therapeutically effective amount of a complex comprising a cardiac targeting peptide linked to a cargo, where the cargo corrects the metabolic defect, where the cardiac targeting peptide has the sequence APWHLSSQYSRT (SEQ ID NO:1).
15. The method of claim 14 , where the metabolic defect is Gaucher's disease and the cargo is glucocerebrosidase.
16. A method of introducing a detectable cargo into a cardiac muscle cell comprising administering, to the cardiac muscle cell, an amount of a complex comprising a cardiac targeting peptide linked to a cargo effective to introduce the cargo into the muscle cell, wherein the cardiac targeting peptide comprises the sequence APWHLSSQYSRT (SEQ ID NO:1), and wherein the cargo comprises a detectable compound.
17. The method of claim 16 , wherein the detectable compound comprises a detectable radioisotope, fluorescent marker, gadolinium marker, or luciferase marker.
18. A method of introducing a detectable cargo into a cardiac muscle cell comprising administering, to the cardiac muscle cell, an amount of a complex comprising a cardiac targeting peptide linked to a cargo effective to introduce the cargo into the muscle cell,
wherein the cardiac targeting peptide
(a) is between 8 and 12 amino acids in length and comprises the sequence HLSSQYSR (SEQ ID NO:2), with the proviso that when the cardiac targeting peptide is 12 amino acids in length it is APWHLSSQX 1 SRT (SEQ ID NO:7) where X 1 is W; or
(b) is between 8 and 12 amino acids in length and comprises the sequence HLSSQYSR (SEQ ID NO:2) in which one amino acid of HLSSQYSR (SEQ ID NO:2) is either deleted or substituted by another amino acid, and
wherein the cargo comprises a detectable compound.
19. The method of claim 18 , wherein the detectable compound comprises a detectable radioisotope, fluorescent marker, gadolinium marker, or luciferase marker.