Modulation of stem and progenitor cell differentiation, assays, and uses thereof

• US 20030235909A1
• Filed: 04/11/2003
• Published: 12/25/2003
• Est. Priority Date: 04/12/2002
• Status: Active Grant

• Current Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company), Celgene Corporation (Bristol-Myers Squibb Company), Signal Pharmaceuticals, LLC (Bristol-Myers Squibb Company)
• Original Assignee: Signal Pharmaceuticals, LLC (Bristol-Myers Squibb Company)
• Inventors: Hariri, Robert J., Stirling, David I., Chan, Kyle W.H., Moutouh-De Parseval, Laure A.
• Granted Patent: US 7,498,171 B2
• Time in Patent Office: Days
• US Class Current: 435/372
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 31/454 containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/14 : of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/02 : Non-specific cardiovascular stimulants, e.g. drugs for syncope, antihypotensives
• 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 2501/125 : Stem cell factor [SCF], c-kit ligand [KL]
• CPC Class Codes: C12N 2501/14 : Erythropoietin [EPO]
• CPC Class Codes: C12N 2501/22 : Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/23 : Interleukins [IL]
• CPC Class Codes: C12N 2501/25 : Tumour necrosing factors [TNF]
• CPC Class Codes: C12N 2501/385 : of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/70 : Enzymes
• CPC Class Codes: C12N 2506/02 : from embryonic cells
• CPC Class Codes: C12N 5/0639 : Dendritic cells, e.g. Langherhans cells in the epidermis
• CPC Class Codes: C12N 5/0642 : Granulocytes, e.g. basopils, eosinophils, neutrophils, mast cells
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

The present invention relates to methods of modulating mammalian stem cell and progenitor cell differentiation. The methods of the invention can be employed to regulate and control the differentiation and maturation of mammalian, particularly human stem cells along specific cell and tissue lineages. The methods of the invention relate to the use of certain small organic molecules to modulate the differentiation of stem or progenitor cell populations along specific cell and tissue lineages, and in particular, to the differentiation of embryonic-like stem cells originating from a postpartum placenta or for the differentiation of early progenitor cells to a granulocytic lineage. Finally, the invention relates to the use of such differentiated stem or progenitor cells in transplantation and other medical treatments.

Claims

1. A method for modulating the differentiation of a mammalian stem cell comprising differentiating said stem cell under suitable conditions and in the presence of a compound that inhibits TNF-α

activity, wherein said compound is not a polypeptide, peptide, protein, hormone, cytokine, oligonucleotide, or nucleic acid.

2. The method of claim 1 wherein said stem cell is differentiated into a hematopoietic cell.

3. The method of claim 1 wherein said stem cell is selected from the group consisting of an embryonic stem cell, a placental stem cell, a cord blood stem cell, a peripheral blood stem cell, and a bone marrow stem cell.

4. The method of claim 1, wherein said compound is an amino-substituted thalidomide analog or amino-substituted isoindoline.

5. The method of claim 4, wherein said amino-substituted thalidomide analog or amino-substituted isoindoline is selected from the group consisting of Actimid™

, Revimid™

, an analog or prodrug of Actimid™

, an analog or prodrug of Revimid™

, and a compound of the formula;

6. The method of claim 4, wherein said amino-substituted thalidomide is selected from the group consisting of 1-oxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-6-aminoisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, 1,3dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, and 1,3dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline.

7. The method of claim 1 wherein the contacting step is conducted in cell culture.

8. The method of claim 1, wherein said contacting step is conducted within said individual.

9. The method of claim 1 wherein the concentration of the compound is from about 0.005 μ

g/ml to about 5 mg/ml.

10. The method of claim 1 wherein the stem cell is a human stem cell.

11. A method for modulating the proliferation or differentiation of a mammalian CD34⁺ or CD 133⁺ progenitor cell comprising proliferating or differentiating said cell under conditions suitable for proliferation or differentiation and in the presence of a compound that inhibits TNF-α

activity, wherein said compound is not a polypeptide, peptide, protein, hormone, cytokine, oligonucleotide, or nucleic acid.

12. The method of claim 11, wherein said progenitor cell is selected from the group consisting of a CD34⁺ progenitor cell and a CD133⁺ progenitor cell.

13. The method of claim 11, wherein said progenitor cells differentiate into CD34⁺CD38⁻

CD33⁺ or CD34⁺CD38⁻

CD33⁻

cells.

14. The method of claim 11, wherein said compound analog is an amino-substituted thalidomide analog or amino-substituted isoindoline.

15. The method of claim 11, wherein said compound is selected from the group consisting of Actimid™

, Revimid™

, an analog or prodrug of Actimid™

, an analog or prodrug of Revimid™

, and a compound of the formula

16. The method of claim 11, wherein said compound is selected from the group consisting of 1-oxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-6-aminoisoindoline, 1-oxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline, 1,3dioxo-2-(2,6-dioxopiperidin-3-yl)-4-aminoisoindoline, and 1,3dioxo-2-(2,6-dioxopiperidin-3-yl)-5-aminoisoindoline.

17. The method of claim 11 wherein the contacting step is conducted in cell culture.

18. The method of claim 11, wherein said contacting step is conducted within an individual.

19. The method of claim 17, wherein said progenitor cells are cells that have been transplanted into said individual.

20. The method of claim 11, wherein said contacting is sufficient to cause a detectable difference in differentiation or proliferation relative to a control.

21. The method of claim 11, wherein said CD34^{+l or CD} 133⁺ progenitor cell has been cryopreserved and thawed prior to said differentiating.

22. A method for expanding a progenitor cell population in a mammalian subject, comprising administering a therapeutically effective amount of CD34⁺ progenitor cells and a compound that inhibits TNF-α

activity to said mammalian subject, wherein said compound is not a polypeptide, peptide, protein, hormone, cytokine, oligonucleotide, or nucleic acid

23. The method of claim 22 wherein said CD34⁺ progenitor cells are differentiated in said mammalian subject.

24. The method of claim 22 wherein said CD34⁺ progenitor cells are administered to said mammalian subject in a cell preparation that is substantially free of red blood cells.

25. The method of claim 22 wherein said CD34⁺ progenitor cells are administered to said mammalian subject in a cell preparation that comprises bone marrow cells, placental cells, or cord blood cells.

26. The method of claim 22 wherein said CD34⁺ progenitor cells are administered to said mammalian subject in conjunction with a carrier.

27. The method of claim 22 wherein said CD34⁺ progenitor cells are CD34⁺CD38⁻

CD33⁺ or CD34⁺CD38⁻

CD33⁻

progenitor cells.

28. The method of claim 22 wherein said CD34⁺ cell is a CD34⁺CD133⁺ progenitor cell.

29. The method of claim 22 wherein the progenitor cells express incorporated genetic material of interest.

30. A pharmaceutical composition comprising a mammalian stem cell and a pharmaceutically-acceptable carrier, wherein said stem cell has been contacted with a compound that inhibits TNF-α

activity for a time sufficient to cause modulation of differentiation or proliferation of said stem cell, and wherein said compound is not a polypeptide, peptide, protein, hormone, cytokine, oligonucleotide, or nucleic acid.

31. The pharmaceutical composition of claim 30 wherein the stem cell is selected from the group consisting of an embryonic stem cell, a placental stem cell, a cord blood stem cell, a peripheral blood stem cell, and a bone marrow stem cell.

32. The pharmaceutical composition of claim 30 wherein said compound is selected from the group consisting of Actimid™

, Revimid™

, an analog or prodrug of Actimid™

, an analog or prodrug of Revimid™

, and a compound of the formula

33. The pharmaceutical composition of claim 30 wherein said contacting step is conducted in cell culture.

34. The pharmaceutical composition of claim 30 wherein the concentration of said compound is from about 0.005 mg/ml to about 5 mg/ml.

35. The pharmaceutical composition of claim 30 wherein the stem cell is a human stem cell.

36. The pharmaceutical composition of claim 30 wherein the differentiation is differentiation into a hematopoietic cell.

37. The pharmaceutical composition of claim 36 wherein said hematopoietic cell is a CD34+ or CD38+ hematopoietic cell.

38. The pharmaceutical composition of claim 36 wherein the hematopoietic cell is a CD11b+ cell.

39. A pharmaceutical composition comprising isolated cord blood cells and an isolated population of white blood cells, wherein the white blood cells are generated by a method comprising differentiating stem cells under suitable conditions and in the presence of a compound that inhibits TNF-α

activity, with the proviso that the compound is not a polypeptide, peptide, protein, hormone, cytokine, oligonucleotide, or nucleic acid, and isolating the white blood cells differentiated thereby.

40. The pharmaceutical composition of claim 39 wherein the compound is an imide or amide.

41. The pharmaceutical composition of claim 39 wherein the differentiating step is conducted in cell culture.

42. The pharmaceutical composition of claim 39 wherein the concentration of the compound is from about 0.005 μ

g/ml to about 5 mg/ml.

43. The pharmaceutical composition of claim 39 wherein the stem cell is a human stem cell.

44. The pharmaceutical composition of claim 39 wherein the stem cell is a progenitor cell.

45. The pharmaceutical composition of claim 44 wherein the progenitor cell is committed to a specific cell lineage.

46. The pharmaceutical composition of claim 44 wherein the progenitor cell is a hematopoietic progenitor cell.

47. A pharmaceutical composition comprising cultured CD34⁺ or CD133⁺ progenitor cells and a pharmaceutically-acceptable carrier, wherein said progenitor cells have been contacted within the first six days of culture with a compound that inhibits the activity of TNF-α

, under conditions that promote proliferation and differentiation of said progenitor cells.

48. The pharmaceutical composition of claim 47 wherein said progenitor cells are collected and cryopreserved after six days of culture.

49. The pharmaceutical composition of claim 47 wherein said progenitor cells are CD34⁺CD38⁻

CD34⁻

or CD34⁺CD38⁻

CD34⁺ cells.

50. The pharmaceutical composition of claim 47 wherein said compound is thalidomide or a thalidomide analog.

51. The pharmaceutical composition of claim 47 in which said compound is Actimid™

or Revimid™

.

52. A method of transplanting a mammalian stem cell comprising:

(a) contacting said stem cell with an immunomodulatory compound to produce a treated stem cell, wherein said contacting is sufficient to modulate the differentiation of said stem cell; and

(b) administering said treated stem cell to an individual.

53. The method of claim 52, wherein step (b) comprises administering said treated stem cell in combination with untreated cells.

54. The method of claim 52 wherein the untreated cell is selected from the group consisting of an embryonic stem cell, a placental cell, a cord blood cell, a peripheral blood cell, and a bone marrow cell.

55. The method of claim 52, wherein said stem cell has been cryopreserved and thawed prior to said administering.

56. A method of transplanting a mammalian progenitor cell comprising:

(a) contacting said progenitor cell with immunomodulatory compound to produce a treated progenitor cell, wherein said contacting is sufficient to modulate the differentiation of said progenitor cell; and

(b) administering said treated progenitor cell to an individual.

57. The method of claim 56, wherein step (b) comprises administering said treated progenitor cell in combination with untreated cells.

58. The method of claim 56 wherein the untreated cell is selected from the group consisting of an embryonic stem cell, a placental cell, a cord blood cell, a peripheral blood cell, and a bone marrow cell.

59. The method of claim 56, wherein said stem cell has been cryopreserved and thawed prior to said administering.

60. A method of treating an individual experiencing a condition comprising administering to said individual an agent selected from the group consisting of:

(a) a compound that inhibits TNF-α

activity, wherein said compound is not a polypeptide, peptide, protein, hormone, cytokine, oligonucleotide, or nucleic acid;

(b) a stem cell differentiated in the presence of said compound; and

(c) a progenitor cell differentiated in the presence of said compound, wherein said agent detectably reduces or ameliorates said condition.

61. The method of claim 60, wherein said condition is selected from the group consisting of inflammation, heart disease, vascular disease, amylotrophic lateral sclerosis, a lysosomal storage disease, and diabetes.

62. The method of claim 60, wherein said agent comprises both a stem cell and compound that inhibits TNF-α

activity, wherein said compound is not a polypeptide, peptide, protein, hormone, cytokine, oligonucleotide, or nucleic acid

63. A method of treating an individual comprising administering a therapeutically effective amount of white blood cells to said recipient mammalian subject, wherein said white blood cells are generated by a method comprising differentiating a stem cell under suitable conditions and in the presence of a compound that inhibits TNF-α

activity, with the proviso that the compound is not a polypeptide, peptide, protein, hormone, cytokine, oligonucleotide, or nucleic acid.

64. The method of claim 63 wherein the stem cells are differentiated in vitro.

65. The method of claim 63 wherein the stem cells are differentiated in a postpartum perfused placenta.

66. The method of claim 63 wherein the white blood cells are administered to the individual in a cell preparation that is substantially free of red blood cells.

67. The method of claim 63 wherein the white blood cells are administered to the individual in a cell preparation which comprises cord blood cells.

68. The method of claim 63 wherein the white blood cells are administered to the individual in conjunction with a carrier.

69. The method of claim 63 wherein the white blood cells are administered to treat or repair a defect in the recipient mammalian subject.

70. The method of claim 69 wherein the defect is a hematopoietic or blood cell proliferation defect.

71. The method of claim 69 wherein the hematopoietic or blood cell proliferation defect is neutropenia or leukopenia.

72. The method of claim 69 wherein the white blood cells are administered systemically.

73. The method of claim 69 wherein the white blood cells are administered intravenously.

74. The method of claim 69 wherein the white blood cells express incorporated genetic material of interest.

75. The method of claim 63 wherein the white blood cells are allogeneic.

76. The method of claim 63 wherein the recipient mammalian subject is human.

77. A method of making a pharmaceutical composition, comprising:

(a) contacting CD34⁺ or CD133⁺ progenitor cells with a compound that inhibits TNF-α

activity, wherein said progenitor cells are cultured for six days under culture conditions that allow proliferation and differentiation of said progenitor cells;

(b) collecting said cells after six days of culture; and

(c) placing said cells in a pharmaceutically-acceptable carrier.