Adult Human Cardiac-Derived Progenitor Cells

• US 20110110897A1
• Filed: 01/09/2009
• Published: 05/12/2011
• Est. Priority Date: 01/11/2008
• Status: Abandoned Application

• Current Assignee: Baylor College of Medicine (Baylor College)
• Original Assignee: Baylor College of Medicine (Baylor College)
• Inventors: Schneider, Michael D., Noseda, Michela, Schwarz, Richard P.
• Application Number:
  US12/812,353
• Publication Number:
  US 20110110897A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• 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: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• 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: C12N 5/0657 Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/0662 Stem cells

Abstract

The present invention is based, in part, on the discovery that cardiac progenitor cells are present in and can be isolated from adult human heart. Accordingly, a cell of the present invention comprises a human adult cardiac-derived progenitor cell capable of differentiating into a cardiac myocyte where said cell is isolated according to the expression of specific biomarkers, identified elsewhere herein. The present invention also includes methods of use of an adult cardiac-derived progenitor cell in the treatment of heart disease.

Claims

1. An isolated human adult progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein said cell expresses an Abcg2 protein or an mRNA encoding Abcg2, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

2. The cell of claim 1, wherein said cell further expresses a mesenchymal stem cell marker protein or a nucleic acid encoding said stem cell marker protein selected from the group consisting of CD105, CD90, and CD44.

3. The cell of claim 1, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

4. The cell of claim 1, wherein said cell is capable of differentiating into a cardiac muscle cell.

5. A pharmaceutical composition comprising a therapeutically effective amount of an isolated human adult progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein said cell expresses an Abcg2 protein or an mRNA encoding Abcg2, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

6. The pharmaceutical composition of claim 5, wherein said cell further expresses a mesenchymal stem cell marker protein or a nucleic acid encoding said stem cell marker protein selected from the group consisting of CD105, CD90, and CD44.

7. The pharmaceutical composition of claim 5, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

8. The pharmaceutical composition of claim 5, wherein said cell is capable of differentiating into a cardiac muscle cell.

9. A cultured human adult progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein said cell expresses an Abcg2 protein or an mRNA encoding Abcg2, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

10. The cell of claim 9, wherein said cell further expresses a mesenchymal stem cell marker protein or a nucleic acid encoding said stem cell marker protein selected from the group consisting of CD105, CD90, and CD44.

11. The cell of claim 9, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

12. The cell of claim 9, wherein said cell is capable of differentiating into a cardiac muscle cell.

13. A pharmaceutical composition comprising a therapeutic amount of a cultured human adult progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein said cell expresses an Abcg2 protein or an mRNA encoding Abcg2, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

14. The pharmaceutical composition of claim 13, wherein said cell further expresses a mesenchymal stem cell marker protein or a nucleic acid encoding said stem cell marker protein selected from the group consisting of CD105, CD90, and CD44.

15. The pharmaceutical composition of claim 13, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

16. The pharmaceutical composition of claim 13, wherein said cell is capable of differentiating into a cardiac muscle cell.

17. A substantially pure population of adult human progenitor cells, wherein said population is derived from a cardiac tissue sample obtained from a human, wherein said population of cells is selected by fluorescence or magnetic cell sorting, and wherein said population of cells expresses an Abcg2 protein or an mRNA encoding Abcg2, wherein said population of cells also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said population of cells does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of selection.

18. The population of adult human progenitor cells of claim 17, wherein said population of cells express at least one mesenchymal stem cell marker protein or a nucleic acid encoding said stem cell marker protein selected from the list consisting of CD105, CD90, and CD44.

19. The population of adult human progenitor cells of claim 17, where said population of cells does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

20. The population of adult human cardiac-derived progenitor cells of claim 17, wherein said population of cells are capable of differentiating into cardiac muscle cells.

21. A composition comprising a therapeutic amount of an isolated adult human progenitor cell, wherein said cell is derived from a cardiac tissue sample obtained from a human, wherein said cell expresses an Abcg2 protein or an mRNA encoding Abcg2, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

22. The composition of claim 21, wherein said composition further comprises a pharmaceutical acceptable carrier.

23. The composition of claim 21, wherein said cell further expresses a mesenchymal stem cell marker protein or a nucleic acid encoding said stem cell marker protein selected from the group consisting of CD 105, CD90, and CD44.

24. The composition of claim 21, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

25. The pharmaceutical composition of claim 21, wherein said cell is capable of differentiating into a cardiac muscle cell.

26. A method of treating a patient with cardiovascular disease, said method comprising the steps of administering to said patient a therapeutically effective amount of a human progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein when said cell is isolated, said cell expresses an Abcg2 protein or an mRNA encoding Abcg2, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding at said least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

27. The method of claim 26, wherein said cell further expresses a mesenchymal stem cell marker protein or a nucleic acid encoding said stem cell marker protein selected from the group consisting of CD105, CD90, and CD44.

28. The method of claim 26, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

29. The method of claim 26, wherein said cardiovascular disease is selected from the list consisting of coronary artery disease, myocardial infarction, ischemic heart disease, and heart failure.

30. The method of claim 26, wherein said cell is admixed with a pharmaceutical acceptable carrier.

31. The method of claim 26, wherein said cell is autologous.

32. A method of treating damaged myocardium in a patient, said method comprising the steps of:

(a) obtaining cells from an adult cardiac tissue sample;

(b) selecting cells one or more times, before or after expanding said cells, to generate a substantially pure population of cardiac progenitor cells that are c-kit^neg and Abcg2^pos at the time of cell selection;

(c) expanding said cells to form a substantially pure population of adult human cardiac progenitor cells;

(d) administering a therapeutically effective amount of said cardiac progenitor cells to said patient.

33. A method of obtaining a population of human cardiac progenitor cells, said method comprising the steps of:

(a) obtaining cells from a cardiac tissue sample;

(b) selecting cells one or more times, before or after expanding said cells, using either fluorescent or magnetic cell sorting techniques, to generate a substantially pure population of cardiac progenitor cells, wherein said cells are substantially c-kit^neg and Abcg2^pos at the time of sorting;

(c) expanding said cells to form a substantially pure population of adult human cardiac progenitor cells.

34. The method of claim 33, wherein the cells are sorted at least once using one or more biomarkers.

35. An isolated human adult progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein said cell expresses at least one mesenchymal stem cell marker protein or nucleic acid encoding said protein selected from the group consisting of CD105, CD90, and CD44, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

36. The cell of claim 35, wherein said cell further expresses an Abcg2 protein or an mRNA encoding Abcg2.

37. The cell of claim 35, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

38. The cell of claim 35, wherein said cell is capable of differentiating into a cardiac muscle cell.

39. A pharmaceutical composition comprising a therapeutically effective amount of an isolated human adult progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein said cell expresses at least one mesenchymal stem cell marker protein or a nucleic acid encoding said protein selected from the group consisting of CD105, CD90, and CD44, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

40. The pharmaceutical composition of claim 39, wherein said cell further expresses an Abcg2 protein or an mRNA encoding Abcg2.

41. The pharmaceutical composition of claim 39, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

42. The pharmaceutical composition of claim 39, wherein said cell is capable of differentiating into a cardiac muscle cell.

43. A cultured human adult progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein said cell expresses at least one mesenchymal stem cell marker protein or a nucleic acid encoding said protein selected from the group consisting of CD105, CD90, and CD44, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

44. The cell of claim 43, wherein said cell further expresses an Abcg2 protein or an mRNA encoding Abcg2.

45. The cell of claim 43, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

46. The cell of claim 43, wherein said cell is capable of differentiating into a cardiac muscle cell.

47. A pharmaceutical composition comprising a therapeutic amount of a cultured human adult progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein said cell expresses at least one mesenchymal stem cell marker protein or a nucleic acid encoding said protein selected from the group consisting of CD105, CD90, and CD44, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

48. The pharmaceutical composition of claim 47, wherein said cell further expresses an Abcg2 protein or an mRNA encoding Abcg2.

49. The pharmaceutical composition of claim 47, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

50. The pharmaceutical composition of claim 47, wherein said cell is capable of differentiating into a cardiac muscle cell.

51. A substantially pure population of adult human progenitor cells, wherein said population is derived from a cardiac tissue sample obtained from a human, wherein said population of cells is selected by fluorescence or magnetic cell sorting, and wherein said population of cells expresses at least one mesenchymal stem cell marker protein or a nucleic acid encoding said protein selected from the list consisting of CD105, CD90, and CD44, wherein said population of cells also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said population of cells does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of selection.

52. The population of adult human progenitor cells of claim 51, wherein said population of cells expresses an Abcg2 protein or an mRNA encoding Abcg2.

53. The population of adult human progenitor cells of claim 51, where said population of cells does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

54. The population of adult human cardiac-derived progenitor cells of claim 51, wherein said population of cells is capable of differentiating into cardiac muscle cells.

55. A composition comprising a therapeutic amount of an isolated adult human progenitor cell, wherein said cell is derived from a cardiac tissue sample obtained from a human, wherein said cell expresses at least one mesenchymal stem cell marker protein or a nucleic acid encoding said protein selected from the group consisting of CD 105, CD90, and CD44, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

56. The composition of claim 55, wherein said composition further comprises a pharmaceutical acceptable carrier.

57. The composition of claim 55, wherein said cell further expresses an Abcg2 protein or an mRNA encoding Abcg2.

58. The composition of claim 55, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

59. The pharmaceutical composition of claim 55, wherein said cell is capable of differentiating into a cardiac muscle cell.

60. A method of treating a patient with heart disease, said method comprising the steps of administering to said patient a therapeutically effective amount of a human progenitor cell, wherein said cell is isolated from a cardiac tissue sample, wherein when said cell is isolated, said cell expresses at least one mesenchymal stem cell marker protein or a nucleic acid encoding said protein selected from the group consisting of CD105, CD90, and CD44, wherein said cell also expresses at least one cardiac transcription factor protein selected from the group consisting of dHAND, Gata4, Mef2a, Tbx2, Tbx5, Tbx20, or an mRNA encoding said at least one cardiac transcription factor, and further wherein, said cell does not substantially express a c-kit protein or an mRNA encoding c-kit at the time of cell isolation.

61. The method of claim 60, wherein said cell further expresses an Abcg2 protein or an mRNA encoding Abcg2.

62. The method of claim 60, where said cell does not express at least one hematopoietic biomarker protein or a nucleic acid encoding said biomarker protein selected from the group consisting of CD45, CD34, CD31, Lin, Lmo2, Gata2, and Tal.

63. The method of claim 60, wherein said cardiovascular disease is selected from the list consisting of coronary artery disease, myocardial infarction, ischemic heart disease, and heart failure.

64. The method of claim 60, wherein said cell is admixed with a pharmaceutical acceptable carrier.

65. The method of claim 60, wherein said cell is autologous.

66. A method of treating damaged myocardium in a patient, said method comprising the steps of:

(a) obtaining cells from an adult cardiac tissue sample;

(b) selecting cells one or more times, before or after expanding said cells, to generate a substantially pure population of cardiac progenitor cells that are c-kit^neg and positive for at least one mesenchymal stem cell marker selected from the list consisting of CD105, CD90, and CD44 at the time of cell selection;

(c) expanding said selected cells to form a substantially pure population of adult human cardiac progenitor cells;

(d) administering a therapeutically effective amount of said cardiac progenitor cells so expanded to said patient.

67. A method of obtaining a population of human cardiac progenitor cells, said method comprising the steps of:

(a) obtaining cells from a cardiac tissue sample;

(b) selecting cells one or more times, before or after expanding said cells, using either fluorescent or magnetic cell sorting techniques, to generate a substantially pure population of cardiac progenitor cells, wherein said cells are substantially c-kit^neg and positive for at least one mesenchymal stem cell marker selected from the list consisting of CD105, CD90, and CD44 at the time of sorting;

(c) expanding said selected cells to form a substantially pure population of adult human cardiac progenitor cells.

68. The method of claim 67, wherein the cells are sorted at least once using one or more biomarkers.

71. The method of claim 67, wherein the cells are sorted at least once using one or more biomarkers.

69. A method of treating damaged myocardium in a patient, said method comprising the steps of:

(a) obtaining cells from an adult cardiac tissue sample;

(b) selecting cells one or more times, before or after expanding said cells, to generate a substantially pure population of cardiac progenitor cells that are c-kit^neg and positive for at least one marker selected from the list consisting of Abcg2, CD 105, CD90, and CD44 at the time of cell selection;

(c) expanding said selected cells to form a substantially pure population of adult human cardiac progenitor cells;

(d) administering a therapeutically effective amount of said cardiac progenitor cells so expanded to said patient.

70. A method of obtaining a population of human cardiac progenitor cells, said method comprising the steps of:

(a) obtaining cells from a cardiac tissue sample;

(b) selecting cells one or more times, before or after expanding said cells, using either fluorescent or magnetic cell sorting techniques, to generate a substantially pure population of cardiac progenitor cells, wherein said cells are substantially c-kit^neg and positive for at least one marker selected from the list consisting of Abcg2, CD105, CD90, and CD44 at the time of sorting;

(c) expanding said selected cells to form a substantially pure population of adult human cardiac progenitor cells.