Treatment of heart disease

• US 10,568,912 B2
• Filed: 11/06/2017
• Issued: 02/25/2020
• Est. Priority Date: 11/09/2009
• Status: Active Grant

• Current Assignee: AAL Scientifics, Inc.
• Original Assignee: AAL Scientifics, Inc.
• Inventors: Anversa, Piero, Leri, Annarosa
• Primary Examiner(s): Berke-Schlessel, David W
• Application Number:
  US15/804,339
• Publication Number:
  US 20180055889A1
• Time in Patent Office: 841 Days
• Field of Search: None
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/34 Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes vascular smooth muscle A61K35/44
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 5/0657 Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/0662 Stem cells

Abstract

Disclosed herein are methods, compositions and kits for treating cardiac stem cells to be administered to a subject in need thereof, e.g., with a damaged myocardium. The methods, composition and kits of the invention can be used to treat cardiovascular diseases such as heart failure, myocardial infarction and an age-related cardiomyopathy.

Claims

1. A method of treating cardiac stem cells comprising:

a. contacting a population of cardiac stem cells with an effective amount of IGF-1 or a variant thereof; and

b. contacting the population of cardiac stem cells from step (a) with an effective amount of IGF-2 or a variant thereof,

wherein the population of cardiac stem cells is isolated from a myocardial tissue of a human subject.

2. The method of claim 1, wherein at least a subset of the population of cardiac stem cells express IGF-1 receptor.

3. The method of claim 1, wherein the effective amount of IGF-1 or a variant thereof is sufficient to increase proliferation of the cardiac stem cells by at least about 10%, as compared to cardiac stem cells in the absence of IGF-1.

4. The method of claim 1, wherein the effective amount of IGF-1 or a variant thereof is about 100 ng/mL.

5. The method of claim 1, wherein the cardiac stem cells are contacted with IGF-1 for at least about 12 hours.

6. The method of claim 1, wherein the effective amount of IGF-2 is sufficient to increase expression of at least one marker for myocyte differentiation by at least about 10%, as compared to cardiac stem cells in the absence of IGF-2.

7. The method of claim 6, wherein the marker for myocyte differentiation is selected from the group consisting of:

α

-sarcomeric actin, connexin-43, Nkx2.5, GATA-4 and MEF2C.

8. The method of claim 1, wherein the effective amount of IGF-2 or a variant thereof is about 100 ng/mL.

9. The method of claim 1, wherein the cardiac stem cells are contacted with IGF-2 for an amount of time sufficient to increase myocyte formation, as compared to cardiac stem cells in the absence of IGF-2.

10. The method of claim 1, wherein the cardiac stem cells are contacted with IGF-2 for an amount of time sufficient for the cardiac stem cells to confer acquisition of at least one adult cardiomyocyte phenotype, as compared to cardiac stem cells in the absence of IGF-2.

11. The method of claim 10, wherein the adult cardiomyocyte phenotype is selected from the group consisting of:

a myocyte volume greater than 2000 μ

m³, contractility in response to electric stimulation, an elongated morphology, calcium tolerance, expression of at least one contractile protein, and activation of ion current channels.

12. The method of claim 11, wherein the contractile protein is selected from the group consisting of:

α

-sarcomeric actin, myosin heavy chains, myosin light chains and tropomyosin.

13. The method of claim 1, wherein the population of cardiac stem cells is administered to an area of a damaged heart tissue of the subject in need thereof.

14. The method of claim 1, wherein the subject in need thereof has a damaged myocardium.

15. The method of claim 1, wherein the subject in need thereof is diagnosed with or suffering from a myocardial infarction.

16. The method of claim 1, wherein the subject in need thereof is diagnosed with or suffering from a heart failure.

17. The method of claim 1, wherein the subject in need thereof is diagnosed with or suffering from an age-related cardiomyopathy.

18. The method of claim 11, wherein the adult cardiomyocyte phenotype is a myocyte volume greater than 2000 μ

m³.

19. The method of claim 11, wherein the adult cardiomyocyte phenotype is contractility in response to electric stimulation.

20. The method of claim 11, wherein the adult cardiomyocyte phenotype is an elongated morphology.

21. The method of claim 11, wherein the adult cardiomyocyte phenotype is calcium tolerance.

22. The method of claim 11, wherein the adult cardiomyocyte phenotype is expression of at least one contractile protein.

23. The method of claim 11, wherein the adult cardiomyocyte phenotype is activation of ion current channels.

24. The method of claim 12, wherein the contractile protein is α

-sarcomeric actin.

25. The method of claim 12, wherein the contractile protein is myosin heavy chain.

26. The method of claim 12, wherein the contractile protein is myosin light chain.

27. The method of claim 12, wherein the contractile protein is tropomyosin.

28. The method of claim 1, wherein the cardiac stem cells express at least one of an IGF-1 receptor, (IGF-1R), an IGF-2 receptor (IGF-2R), and an angiotensin type 1 receptor (AT1R), and combinations thereof.

29. The method of claim 1, wherein the cardiac stem cells are lineage negative.

30. The method of claim 1, wherein the cardiac stem cells express c-kit.

31. The method of claim 1, wherein the cardiac stem cells are lineage negative and express c-kit.

32. The method of claim 1, wherein the population of cardiac stem cells is pre-selected for expression of IGF-1R.

33. The method of claim 1, wherein the population of cardiac stem cells is pre-selected for expression of IGF-2R.

34. The method of claim 1, wherein the population of cardiac stem cells is pre-selected for expression of IGF-1R and IGF-2R.