Muscle-derived cells (MDCs) for promoting and enhancing nerve repair and regeneration

• US 20050238625A1
• Filed: 04/26/2004
• Published: 10/27/2005
• Est. Priority Date: 04/25/2003
• Status: Active Grant

• Current Assignee: University Of Pittsburgh (University of Pittsburgh of The Commonwealth System of Higher Education)
• Original Assignee: University Of Pittsburgh (University of Pittsburgh of The Commonwealth System of Higher Education)
• Inventors: Minnery, Brandon, Chancellor, Michael B., Huard, Johnny
• Granted Patent: US 9,617,516 B2
• Time in Patent Office: Days
• US Class Current: 424/93.210
• 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 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: C12N 5/0658 Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts

Abstract

The present invention describes methods involving the use of muscle derived cells (MDCs), preferably obtained from skeletal muscle, to support the innervation and repair of damaged tissues and organs, particularly associated with nerve damage or neuropathy. The invention relates to MDCs for use in methods for promoting or enhancing innervation of nerve cells, particularly in the peripheral nervous system, and their ability to contribute to the development of neuronal tissue when MDCs are introduced at or near a tissue or organ site in need of repair due to injury, damage, disease, or dysfunction. Such methods are useful for the treatment of central and peripheral nervous system disorders and to alleviate, abate, or eliminate the symptoms of neurologic or neurodegenerative diseases in animals, particularly mammals, including humans. The methods are also useful for treating both nerve and muscle tissue following injury, damage, or dysfunction to these tissue types.

Claims

1. A method of promoting or enhancing survival, regeneration, and/or innervation of damaged, injured, diseased, or dysfunctional nerve tissue, comprising:

introducing muscle-derived cells (MDCs) in or around a site of tissue damage, injury, disease, or dysfunction in a recipient, in an amount effective to promote or enhance the survival, regeneration, and/or innervation of the damaged, injured, diseased, or dysfunctional nerve tissue.

2. The method according to claim 1, wherein the MDCs are autologous to the tissue into which they are introduced.

3. The method according to claim 1, wherein the MDCs are histocompatibly-matched with the recipient in need of treatment.

4. The method according to claim 1, wherein the MDCs are obtained from skeletal muscle tissue.

5. The method according to claim 1, wherein the MDCs are introduced in an amount of about 10⁵ to 10⁶ cells per cm³ of tissue to be treated in a physiologically acceptable medium.

6. The method according to claim 1, wherein a cloned population of the MDCs is introduced into the recipient.

7. The method according to claim 1, wherein the MDCs carry at least one heterologous polynucleotide encoding a heterologous protein or polypeptide for promoting or enhancing innervation.

8. The method according to claim 7, wherein the protein or polypeptide is selected from at least one of a growth factor, growth factor receptor, peptide neurotransmitter and enzyme involved in the synthesis of neurotransmitters.

9. The method according to claim 8, wherein the growth factor is selected from nerve growth factor (NGF), ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF), neurotrophin 3, neurotrophin 4, interleukins, fibroblast growth factor 1 (FGF-1), fibroblast growth factor 1 (FGF-2), insulin-like growth factors (IGF), glucocorticoid hormone, somatostatin, platelet-derived growth factor (PDGF), tetanus toxoid, TGF-alpha, TGF-beta, epidermal growth factor, or retinoic acid.

10. The method according to claim 9, wherein the protein or polypeptide is nerve growth factor (NGF).

11. The method according to claim 8, wherein the growth factor receptor is selected from trk neurotrophin receptor, nerve growth factor (NGF) receptor, ciliary neurotrophic factor (CNTF) receptor, brain-derived neurotrophic factor (BDNF) receptor, neurotrophin 3 receptor, neurotrophin 4 receptor, interleukin receptor, fibroblast growth factor 1 (FGF-1) receptor, fibroblast growth factor 1 (FGF-2) receptor, insulin-like growth factor (IGF) receptor, glucocorticoid hormone receptor, platelet-derived growth factor (PDGF) receptor, TGF-alpha receptor, TGF-beta receptor, epidermal growth factor (EGF) receptor, or retinoic acid receptor.

12. The method according to claim 7, wherein the MDCs are transduced with a viral vector containing the at least one heterologous polynucleotide.

13. The method according to claim 7, wherein the MDCs are transfected with plasmid DNA containing the at least one heterologous polynucleotide.

14. The method according to claim 12, wherein the viral vector is selected from the group consisting of adenovirus, herpes simplex virus, adeno-associated virus, and retrovirus.

15. The method according to claim 12, wherein the viral vector is a replication-defective viral vector selected from the group consisting of adenovirus, herpes simplex virus, adeno-associated virus and retrovirus.

16. The method according to claim 1, wherein the tissue injury, damage, disease, or dysfunction comprises a disease, disorder, or condition selected from stroke, spinal injury, multiple sclerosis, amyotrophic lateral sclerosis, epilepsy, Parkinson'"'"'s Disease, Huntington'"'"'s Disease, Alzheimer'"'"'s Disease, ischemia following cardiac arrest, diabetes, peripheral nerve injury, peripheral nerve inflammation, autonomic nerve injury, pelvic nerve damage, burn, blunt trauma, back injury, back pain, or sciatica.

17. The method according to claim 1, wherein the MDCs are enriched from a muscle cell suspension by plating the suspension into at least two successive cultures so as to eliminate fibroblasts and non-muscle cells and to enrich for an end population of mononucleated MDCs.

18. The method according to claim 17, wherein the enriched end population of mononucleated MDCs comprises myoblasts.

19. The method according to claim 1, wherein the MDCs are administered in combination with a physiologically acceptable carrier, excipient, or diluent.

20. A method of repairing peripheral nerve and muscle tissue at the same tissue site or location in a recipient in need thereof, comprising:

introducing muscle-derived cells (MDCs) at the tissue site or location, in an amount effective for MDCs to regenerate and repopulate the tissue site to repair both the nerve and muscle tissue.

21. The method according to claim 20, wherein the MDCs are autologous to the nerve tissue into which they are introduced.

22. The method according to claim 20, wherein the MDCs are histocompatibly-matched with the recipient in need of treatment.

23. The method according to claim 20, wherein the MDCs are obtained from skeletal muscle tissue.

24. The method according to claim 20, wherein the MDCs are introduced in an amount of about 10⁵ to 10⁶ cells per cm³ of tissue to be treated in a physiologically acceptable medium.

25. The method according to claim 20, wherein a cloned population of the MDCs is introduced into the recipient.

26. The method according to claim 20, wherein the MDCs carry at least one heterologous polynucleotide encoding a heterologous protein or polypeptide for promoting or enhancing innervation.

27. The method according to claim 26, wherein the protein or polypeptide is selected from at least one of a growth factor, growth factor receptor, peptide neurotransmitter and enzyme involved in the synthesis of neurotransmitters.

28. The method according to claim 27, wherein the growth factor is selected from nerve growth factor (NGF), ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF), neurotrophin 3, neurotrophin 4, interleukins, fibroblast growth factor 1 (FGF-1), fibroblast growth factor 1 (FGF-2), insulin-like growth factors (IGF), glucocorticoid hormone, somatostatin, platelet-derived growth factor (PDGF), tetanus toxoid, TGF-alpha, TGF-beta, epidermal growth factor, or retinoic acid.

29. The method according to claim 26, wherein the protein or polypeptide is nerve growth factor (NGF).

30. The method according to claim 27, wherein the growth factor receptor is selected from trk neurotrophin receptor, nerve growth factor (NGF) receptor, ciliary neurotrophic factor (CNTF) receptor, brain-derived neurotrophic factor (BDNF) receptor, neurotrophin 3 receptor, neurotrophin 4 receptor, interleukin receptor, fibroblast growth factor 1 (FGF-1) receptor, fibroblast growth factor 1 (FGF-2) receptor, insulin-like growth factor (IGF) receptor, glucocorticoid hormone receptor, platelet-derived growth factor (PDGF) receptor, TGF-alpha receptor, TGF-beta receptor, epidermal growth factor (EGF) receptor, or retinoic acid receptor.

31. The method according to claim 26, wherein the MDCs are transduced with a viral vector containing the at least one heterologous polynucleotide.

32. The method according to claim 26, wherein the MDCs are transfected with plasmid DNA containing the at least one heterologous polynucleotide.

33. The method according to claim 31, wherein the viral vector is selected from the group consisting of adenovirus, herpes simplex virus, adeno-associated virus, and retrovirus.

34. The method according to claim 31, wherein the viral vector is a replication-defective viral vector selected from the group consisting of adenovirus, herpes simplex virus, adeno-associated virus and retrovirus.

35. The method according to claim 20, wherein the tissue injury, damage, disease, or dysfunction comprises a disease, disorder, or condition selected from stroke, spinal injury, multiple sclerosis, amyotrophic lateral sclerosis, epilepsy, Parkinson'"'"'s Disease, Huntington'"'"'s Disease, Alzheimer'"'"'s Disease, ischemia following cardiac arrest, diabetes, peripheral nerve injury, peripheral nerve inflammation, autonomic nerve injury, pelvic nerve damage, burn, blunt trauma, back injury, back pain, or sciatica.

36. The method according to claim 20, wherein the MDCs are enriched from a muscle cell suspension by plating the suspension into at least two successive cultures so as to eliminate fibroblasts and non-muscle cells and to enrich for an end population of mononucleated MDCs.

37. The method according to claim 36, wherein the enriched end population of mononucleated MDCs comprises myoblasts.

38. The method according to claim 20, wherein the MDCs are administered in combination with a physiologically acceptable carrier, excipient, or diluent.

39. A method of promoting or enhancing innervation of a tissue or organ transplant or graft in a host tissue or organ, comprising:

introducing muscle-derived cells (MDCs) into an area comprising a boundary between the tissue or organ transplant or graft, and the host tissue or organ in a recipient, in an amount effective for said MDCs to promote or enhance innervation between the host tissue or organ and the tissue or organ transplant.

40. The method according to claim 39, wherein the MDCs facilitate survival and growth of nerve fibers between the tissue or organ transplant or graft, and the host tissue or organ.

41. The method according to claim 39, wherein the MDCs are autologous to either (i) the host tissue or organ or (ii) the tissue or organ transplant or graft.

42. The method according to claim 39, wherein the tissue or organ transplant or graft is selected from one or more of digestive, reproductive, cardiovascular, urological, respiratory, epithelial, connective, neuronal, endocrine, skin, smooth muscle, skeletal muscle tissues or organs, or a portion or section thereof.

43. The method according to claim 39, wherein the tissue or organ transplant or graft comprises a skin substitute or engineered material having properties mimetic of natural tissue.

44. The method according to claim 43, wherein the skin substitute or engineered material comprises one or more of acellular cadaveric skin matrix, poly-lactic acid, hyaluronic acid, poly-glycolic acid, polyethylene oxide, polybutylene terephthalate, silicone, autologous cultured fibroblasts, allogeneic cultured fibroblasts, autologous cultured keratinocytes, allogeneic cultured keratinocytes, autologous cultured epidermis, allogeneic cultured epidermis, autologous cultured epithelium, allogeneic cultured epithelium, de-epidermized dermis, collagen sponge, collagen-chitosan sponge, chitosan-cross-linked collagen-glycosaminoglycan matrix, collagen gel, polyglactin mesh, or small intestinal submucosa (SIS).

45. The method according to claim 43, wherein the skin substitute or engineered material is introduced in combination with dermal tissue or dermal tissue components.

46. The method according to claim 45, wherein the dermal tissue or dermal tissue component is selected from one or more of keratinocytes, fibroblasts, melanocytes, dendritic cells, adnexal cells, neuronal cells, glial cells, endothelial cells, or smooth muscle cells.

47. The method according to claim 39, wherein the MDCs carry at least one heterologous polynucleotide encoding a heterologous protein or polypeptide for enhancing the innervation of transplanted tissues or organs.

48. The method according to claim 47, wherein the protein or polypeptide is selected from at least one of a growth factor, growth factor receptor, peptide neurotransmitter, or enzyme involved in the synthesis of neurotransmitters.

49. The method according to claim 48, wherein the growth factor is selected from nerve growth factor (NGF), ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF), neurotrophin 3, neurotrophin 4, interleukins, fibroblast growth factor 1 (FGF-1), fibroblast growth factor 1 (FGF-2), insulin-like growth factors (IGF), glucocorticoid hormone, somatostatin, platelet-derived growth factor (PDGF), tetanus toxoid, TGF-alpha, TGF-beta, epidermal growth factor, or retinoic acid.

50. The method according to claim 49, wherein the protein or polypeptide is nerve growth factor (NGF).

51. The method according to claim 48, wherein the growth factor receptor is selected from trk family of neurotrophin receptors, nerve growth factor (NGF) receptor, ciliary neurotrophic factor (CNTF) receptor, brain-derived neurotrophic factor (BDNF) receptor, neurotrophin 3 receptor, neurotrophin 4 receptor, interleukin receptor, fibroblast growth factor 1 (FGF-1) receptor, fibroblast growth factor 1 (FGF-2) receptor, insulin-like growth factor (IGF) receptor, glucocorticoid hormone receptor, platelet-derived growth factor (PDGF) receptor, TGF-alpha receptor, TGF-beta receptor, epidermal growth factor (EGF) receptor, or retinoic acid receptor.

52. The method according to claim 39, wherein the MDCs are enriched from a muscle cell suspension by plating the suspension into at least two successive cultures so as to eliminate fibroblasts and non-muscle cells and to enrich for an end population of mononucleated MDCs.

53. The method according to claim 52, wherein the enriched end population of mononucleated MDCs comprises myoblasts.

54. The method according to claim 39, wherein the MDCs are histocompatibly-matched with the recipient in need of treatment.

55. The method according to claim 39, wherein the MDCs are obtained from skeletal muscle tissue.

56. The method according to claim 39, wherein the MDCs are introduced in an amount of about 10⁵ to 10⁶ cells per cm³ of tissue or organ in a physiologically acceptable medium.

57. The method according to claim 39, wherein a cloned population of the MDCs is introduced into the recipient.

58. A method of promoting or enhancing innervation of a severed or partially severed tissue or limb, comprising:

introducing muscle-derived cells (MDCs) into a site of the severed or partially severed tissue or limb of a subject, in an amount effective for said MDCs to promote or enhance innervation in the tissue or limb.

59. The method according to claim 58, wherein the MDCs are introduced at one or more of the distal site, the proximal site, or both the distal site and the proximal site of the severed tissue or limb.

60. The method according to claim 58, wherein the MDCs are administered either separately or in combination with a physiologically acceptable carrier, excipient, or diluent.

61. The method according to claim 58, where the MDCs are engineered to carry at least one heterologous polynucleotide encoding a heterologous protein or polypeptide for enhancing the innervation of transplanted tissues or organs.

62. The method according to claim 61, wherein the protein or polypeptide is selected from at least one of a growth factor, growth factor receptor, peptide neurotransmitter, or enzyme involved in the synthesis of neurotransmitters.

63. The method according to claim 58, wherein the MDCs are enriched from a muscle cell suspension by plating the suspension into at least two successive cultures so as to eliminate fibroblasts and non-muscle cells and to enrich for an end population of mononucleated MDCs.

64. The method according to claim 63, wherein the enriched end population of mononucleated MDCs comprises myoblasts.

65. The method according to claim 58, wherein the MDCs are histocompatibly-matched with the recipient in need of treatment.

66. The method according to claim 58, wherein the MDCs are obtained from skeletal muscle tissue.

67. The method according to claim 58, wherein the MDCs are introduced in an amount of about 10⁵ to 10⁶ cells per cm³ of tissue or organ in a physiologically acceptable medium.

68. The method according to claim 58, wherein a cloned population of the MDCs is introduced into the recipient.

69. A method of promoting or enhancing neurorecovery and recovery of erectile function following prostatectomy, comprising:

administering to a patient in need thereof autologous muscle-derived cells (MDCs) in an amount effective to promote or enhance neurorecovery and the recovery of erectile function in the patient.

70. The method according to claim 69, wherein the MDCs are administered at one or more sites of prostatectomy anastomosis.

71. The method according to claim 69, wherein the MDCs are administered in or around the cavernous nerves, pelvic nerves, hypogastric nerves, pelvic plexi, and pelvic autonomic ganglia.

72. The method according to claim 69, wherein the MDCs are administered into the penis.

73. The method according to claim 69, wherein the prostatectomy is radical prostatectomy.

74. The method according to claim 69, further comprising cytoscopic re-administration of MDCs to the patient following surgery.

75. The method according to claim 69, wherein the MDCs are enriched from a muscle cell suspension by plating the suspension into at least two successive cultures so as to eliminate fibroblasts and non-muscle cells and to enrich for an end population of mononucleated MDCs.

76. The method according to claim 75, wherein the enriched end population of mononucleated MDCs comprises myoblasts.

77. The method according to claim 69, wherein the MDCs are obtained from skeletal muscle tissue.

78. The method according to claim 69, wherein the MDCs are administered in an amount of about 10⁵ to 10⁶ cells per cm³ of tissue of the patient.

79. The method according to claim 69, wherein the MDCs are administered in conjunction with a physiologically acceptable carrier, diluent, or excipient.

80. The method according to claim 69, wherein a cloned population of the MDCs is administered to the patient.

81. A method of preventing or retarding degeneration of peripheral cutaneous axons following tissue damage, comprising:

introducing muscle-derived cells (MDCs) into the damaged tissue in a recipient, in an amount effective for said MDCs to prevent or retard the degeneration of peripheral cutaneous axons.

82. The method according to claim 81, wherein the peripheral cutaneous axons have been damaged as a result of tissue injury, disease, or dysfunction.

83. The method according to claim 82, wherein the tissue injury, disease, or dysfunction is selected from inflammation, burn, trauma, blunt trauma, or insult.

84. A method of promoting or enhancing axonal regeneration and/or neuronal survival at the site of nerve damage, disease, or injury, comprising:

introducing muscle-derived cells (MDCs) into the site of tissue damage, disease, or injury in a recipient, in an amount effective for said MDCs to promote or enhance axonal regeneration and/or neuronal survival.

85. The method according to claim 84, wherein the MDCs are enriched from a muscle cell suspension by plating the suspension into at least two successive cultures so as to eliminate fibroblasts and non-muscle cells and to enrich for an end population of mononucleated MDCs.

86. The method according to claim 85, wherein the enriched end population of mononucleated MDCs comprises myoblasts.

87. The method according to claim 84, wherein the MDCs are obtained from skeletal muscle tissue.

88. A method of promoting or enhancing neurorecovery and repair of pelvic nerves damaged during prostatectomy or pelvic surgery, comprising:

administering to a patient in need thereof autologous muscle-derived cells (MDCs) in an amount effective to promote or enhance the neurorecovery and repair of pelvic nerves damaged during prostatectomy or pelvic surgery in the patient.

89. The method according to claim 88, wherein the MDCs are enriched from a muscle cell suspension by plating the suspension into at least two successive cultures so as to eliminate fibroblasts and non-muscle cells and to enrich for an end population of mononucleated MDCs.

90. The method according to claim 89, wherein the enriched end population of mononucleated MDCs comprises myoblasts.

91. The method according to claim 88, wherein the MDCs are obtained from skeletal muscle tissue.

92. The method according to claim 88, wherein the MDCs are administered in an amount of about 10⁵ to 10⁶ cells per cm³ of tissue of the patient.

93. The method according to claim 88, wherein the MDCs are administered in conjunction with a physiologically acceptable carrier, diluent, or excipient.

94. The method according to claim 88, wherein a cloned population of the MDCs is administered to the patient.

95. The method according to claim 88, wherein the prostatectomy is radical prostatectomy.

96. A method of evaluating the effect of a test substance on the ability of muscle-derived cells (MDCs) to support innervation or nerve cell growth, comprising:

(a) treating a culture of MDCs with the test substance;

(b) introducing the treated MDCs into a site of nerve tissue injury, disease, or damage; and

(c) comparing the ability of the treated MDCs to support innervation or nerve cell growth in the tissue of a recipient having nerve tissue disease, damage, or injury relative to the ability of non-treated MDCs to support innervation or nerve cell growth in the recipient tissue.

97. The method according to claim 96, wherein the MDCs are enriched from a muscle cell suspension by plating the suspension into at least two successive cultures so as to eliminate fibroblasts and non-muscle cells and to enrich for an end population of mononucleated MDCs.