METHODS FOR TREATING ISCHEMIC INJURY

• US 20190328836A1
• Filed: 01/09/2018
• Published: 10/31/2019
• Est. Priority Date: 01/09/2017
• Status: Active Application

• Current Assignee: Duke University, East Carolina University (University of North Carolina System)
• Original Assignee: Duke University, East Carolina University (University of North Carolina System)
• Inventors: Kontos, Christopher D., McClung, Joseph M.
• Application Number:
  US16/476,791
• Publication Number:
  US 20190328836A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/43 containing a FLAG-tag
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: G01N 2800/324 Coronary artery diseases, e.g. angina pectoris, myocardial infarction

Abstract

Embodiments of the present disclosure relate generally to the treatment and prevention of ischemic injury. More particularly, embodiments of the present disclosure include materials and methods for treating and preventing ischemic injury caused by diseases like peripheral artery disease (PAD) by modulating various candidate genes within the Lsq-1 QTL, such as Bcl-2-associated athanogene-3 (Bag3). Given that no alleles that modulate a subject'"'"'s susceptibility to conditions like critical limb ischemia are presently known, there is a need to identify the genetic determinants that play important roles in the treatment and prevention of ischemic injury.

Claims

1. A method of treating ischemic injury in a subject, the method comprising:

administering a polynucleotide encoding a Bcl2-associated athanogene-3 (BAG3) polypeptide to the subject, wherein the BAG3 polypeptide encoded by the polynucleotide is represented by SEQ ID NO;

2; and

treating at least one symptom associated with the ischemic injury in the subject.

2. The method of claim 1, wherein the BAG3 polypeptide encoded by the polynucleotide comprises an isoleucine at amino acid position 79.

3. The method of claim 2, wherein the BAG3 polypeptide comprises at least one amino acid substitution in addition to the isoleucine at amino acid position 79.

4. The method of any of claims 1 to 3, wherein the polynucleotide encoding the BAG3 polypeptide is operably coupled to a targeting vector capable of causing the expression of the BAG3 polypeptide in at least one of a muscle cell, fibroblast, stem cell, pericyte, and endothelial cell.

5. The method of claim 4, wherein the targeting vector comprises an adeno-associated virus (AAV).

6. The method of claim any of claims 1 to 5, wherein the subject has at least one of the following single nucleotide polymorphisms:

(i) a leucine at amino acid position 209;

(ii) an alanine at amino acid position 63;

(iii) a serine at amino acid position 380; and

wherein administering the polynucleotide encoding the BAG3 polypeptide represented by SEQ ID NO;

2 and/or the polynucleotide encoding the BAG3 polypeptide comprising an isoleucine at amino acid position 79 treats the at least one symptom associated with the ischemic injury in the subject.

7. The method of any of claims 1 to 6, wherein the ischemic injury comprises myopathy or vascular deficiency.

8. The method of any of claims 1 to 7, wherein the ischemic injury is caused by one or more of peripheral artery disease comprising intermittent claudication or critical limb ischemia, muscular dystrophy, myofibrillar myopathy, degenerative myopathies, glycogen storage diseases, trauma, renal disease, atrial fibrillation, COPD, coronary artery disease, morbid obesity, cachexia, congestive heart failure, myocardial infarction, and diabetes mellitus.

9. The method of any of claims 1 to 8, wherein at least one symptom is necrosis, and wherein administering the polynucleotide encoding the BAG3 polypeptide treats the ischemic injury in the subject by reducing necrosis.

10. The method of any of claims 1 to 8, wherein at least one symptom is myopathy, and wherein administering the polynucleotide encoding the BAG3 polypeptide treats the ischemic injury in the subject by reducing the myopathy.

11. The method of any of claims 1 to 10, wherein administering the polynucleotide encoding the BAG3 polypeptide represented by SEQ ID NO:

2 and/or the polynucleotide encoding the BAG3 polypeptide comprising an isoleucine at amino acid position 79 treats the at least one symptom associated with the ischemic injury by increasing one or more of muscle fiber cross-sectional area, capillary density, muscle function, muscle regeneration, stem cell activity, vascular density, and vascular luminal diameter.

12. The method of any of claims 1 to 11, wherein administering the polynucleotide encoding the BAG3 polypeptide represented by SEQ ID NO:

2 and/or the polynucleotide encoding the BAG3 polypeptide comprising an isoleucine at amino acid position 79 treats the at least one symptom associated with the ischemic injury by causing one or more of increase in myotube diameter, myotube phenotype, contractile function, an increase in stem cell or satellite cell activity/myogenesis, an increase in mitochondrial number or respiratory function, an increase in autophagic flux, and decreased DNA fragmentation.

13. The method of any of claims 1 to 12, wherein administering the polynucleotide encoding the BAG3 polypeptide represented by SEQ ID NO:

2 and/or the polynucleotide encoding the BAG3 polypeptide comprising an isoleucine at amino acid position 79 treats the at least one symptom associated with the ischemic injury by causing one or more of increased expression of vascular endothelial growth factor (VEGF), neuropilin (Nrp-1), vascular endothelial growth factor receptor 1 (Flt), vascular endothelial growth factor receptor 2 (Flk), myogenin, myoD, Tmem8c (myomaker) and muscle RING-finger protein 1 (MuRF-1), and decreased in expression of myostatin.

14. The method of any of claims 1 to 13, wherein administering the polynucleotide encoding the BAG3 polypeptide represented by SEQ ID NO:

2 and/or the polynucleotide encoding the BAG3 polypeptide comprising an isoleucine at amino acid position 79 to the subject comprises one or more of intramuscular injection, percutaneous injection, intraperitoneal injection, intravenous injection, and oral consumption.

15. The method of any of claims 1 to 14, wherein administering the polynucleotide encoding the BAG3 polypeptide represented by SEQ ID NO:

2 and/or the polynucleotide encoding the BAG3 polypeptide comprising an isoleucine at amino acid position 79 to the subject comprises two or more separate injections.

16. A method of preventing ischemic injury in a subject, the method comprising:

administering a polynucleotide encoding a Bcl2-associated athanogene-3 (BAG3) polypeptide to the subject, wherein the BAG3 polypeptide encoded by the polynucleotide is represented by SEQ ID NO;

2; and

preventing the onset of at least one symptom associated with the ischemic injury in the subject.

17. The method of claim 16, wherein the BAG3 polypeptide encoded by the polynucleotide comprises an isoleucine at amino acid position 79.

18. The method of claim 17, wherein the BAG3 polypeptide comprises at least one amino acid substitution in addition to the isoleucine at amino acid position 79.

19. The method of any of claims 16 to 18, wherein the polynucleotide encoding the BAG3 polypeptide is operably coupled to a targeting vector capable of causing the expression of the BAG3 polypeptide in at least one of a muscle cell, fibroblast, stem cell, pericyte, and endothelial cell.

20. The method of claim any of claims 16 to 19, wherein the subject has at least one of the following single nucleotide polymorphisms:

(i) a leucine at amino acid position 209;

(ii) an alanine at amino acid position 63;

(iii) a serine at amino acid position 380; and

wherein administering the polynucleotide encoding the BAG3 polypeptide represented by SEQ ID NO;

2 and/or the polynucleotide encoding the BAG3 polypeptide comprising an isoleucine at amino acid position 79 prevents the onset of the at least one symptom associated with the ischemic injury in the subject.

21. A pharmaceutical composition for treating or preventing ischemic injury in a subject, the composition comprising:

a polynucleotide encoding a Bcl2-associated athanogene-3 (BAG3) polypeptide, wherein the BAG3 polypeptide encoded by the polynucleotide is represented by SEQ ID NO;

2; and

a pharmaceutically acceptable excipient.

22. The composition of claim 21, wherein the BAG3 polypeptide encoded by the polynucleotide comprises an isoleucine at amino acid position 79.

23. The composition of claim 22, wherein the BAG3 polypeptide comprises at least one amino acid substitution in addition to the isoleucine at amino acid position 79.

24. The composition of any of claims 21 to 23, wherein the polynucleotide encoding the BAG3 polypeptide is operably coupled to a targeting vector capable of causing the expression of the BAG3 polypeptide in at least one of a muscle cell, fibroblast, stem cell, pericyte, and endothelial cell.

25. The composition of any of claims 21 to 24, wherein the composition is formulated for administration to a subject by one or more of intramuscular injection, percutaneous injection, intraperitoneal injection, ingestion, and intravenous injection.

26. A kit comprising:

a pharmaceutical composition comprising a polynucleotide encoding a Bcl2-associated athanogene-3 (BAG3) polypeptide, wherein the BAG3 polypeptide encoded by the polynucleotide is represented by SEQ ID NO;

2; and

a delivery device for administering the pharmaceutical composition to a subject.

27. The kit of claim 26, wherein the BAG3 polypeptide encoded by the polynucleotide comprises an isoleucine at amino acid position 79.

28. The kit of claim 27, wherein the BAG3 polypeptide comprises at least one amino acid substitution in addition to the isoleucine at amino acid position 79.