Isolated stromal cells and methods of using the same

• US 20040166097A1
• Filed: 02/26/2004
• Published: 08/26/2004
• Est. Priority Date: 03/28/1995
• Status: Abandoned Application

• Current Assignee: Thomas Jefferson University (Main Line Health System)
• Original Assignee: Thomas Jefferson University (Main Line Health System)
• Inventors: OʼHara, Michael D., Prockop, Darwin J., Pereira, Ruth F., Laptev, Alexey, Kulkosky, Joseph, Phinney, Donald, Leeper, Dennis B., Caro, Jose
• Application Number:
  US10/787,506
• Publication Number:
  US 20040166097A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 48/0025 wherein the non-active part clearly interacts with the delivered nucleic acid
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 14/745 Blood coagulation or fibrinolysis factors
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C12N 2502/1394 Bone marrow stromal cells; whole marrow
• CPC Class Codes: C12N 2799/027 where the vector is derived from a retrovirus
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

Methods of treating patients who are suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect are disclosed. The methods comprising the step of intravenous administration of stromal cells isolated from normal syngeneic individuals or intravenous administration of stromal cells isolated from the patient subsequent to correction of the genetic defect in the isolated cells. Implant devices comprising a container that has at least one membrane surface and stromal cells isolated from bone marrow that comprise a gene construct are disclosed. The gene construct in the stromal cells comprises a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in stromal cells. Methods of treating individuals with diseases, disorders or conditions which can be treated with a beneficial protein, including diseases, disorders or conditions characterized by gene defects are disclosed. The methods comprise introducing into such individuals, stromal cells that are administered in a manner that physically isolates them from the recipient'"'"'s immune system and that comprise a gene construct that comprises a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in stromal cells.

Claims

1. A method of treating patient who is suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect comprising the steps of:

a) obtaining a bone marrow sample from a donor who is not suffering from a disease, disorder or condition characterized by a bone, cartilage or lung defect and who is syngeneic with said patient;

b) isolating stromal cells from said sample; and

,

c) administering said isolated stromal cells by intravenous infusion to said patient.

2. The method of claim 1 wherein said patent undergoes bone marrow ablation prior to administration of isolated stromal cells.

3. The method of claim 2 wherein said stromal cells are administered by intravenous infusion to said patient together with hematopoeitic precursor cells from a bone marrow sample from a donor who is not suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect and who is syngeneic with said patient.

4. The method of claim 2 wherein said stromal cells are administered by intravenous infusion to said patient free from hematopoeitic precursor cells.

5. The method of claim 1 wherein prior to administering said stromal cells, said stromal cells are transfected with a gene construct that comprises a herpes thymidine kinase gene, wherein said gene is operably linked to regulatory sequences and is expressed by said stromal cells.

6. The method of claim 1 wherein said disease, disorder or condition is characterized by a defect in said patient'"'"'s bone.

7. The method of claim 6 wherein said disease, disorder or condition is osteogenesis imperfecta or osteoporosis.

8. The method of claim 1 wherein said disease, disorder or condition is characterized by a defect in said patient'"'"'s cartilage.

9. The method of claim 8 wherein said disease, disorder or condition is chondrodysplasia or osteoarthritis.

10. The method of claim 1 wherein said disease, disorder or condition is characterized by defect in said patient'"'"'s lungs.

11. The method of claim 10 wherein said disease, disorder or condition characterized is cystic fibrosis.

12. A method of treating patient who suffering from a disease, disorder or condition characterized by a mutated, non-functioning or under-expressed gene which results in a defect in the bone, cartilage or lungs of said patient comprising the steps of:

a) obtaining a bone marrow sample from said patient;

b) isolating stromal cells from said sample;

c) transfecting said stromal cells with a normal copy of said mutated, non-functioning or under-expressed gene wherein said copy of said gene is operably linked to functional regulatory elements; and

d) administering said transfected stromal cells to said patient by intravenous infusion.

13. The method of claim 12 wherein said patent undergoes bone marrow ablation prior to administration of stromal cells.

14. The method of claim 13 wherein said stromal cells are administered by intravenous infusion to said patient together with hematopoietic precursor cells from said sample.

15. The method of claim 12 wherein prior to administering said stromal cells, said stromal cells are transfected with a gene construct that comprises a herpes thymidine kinase gene, wherein said gene is operably linked to regulatory sequences and is expressed by said stromal cells.

16. The method of claim 12 wherein said disease, disorder or condition is characterized by a defect in said patient'"'"'s bone.

17. The method of claim 16 wherein said disease, disorder or condition is osteogenesis imperfecta and said gene encodes type I procollagen or type I collagen.

18. The method of claim 12 wherein said disease, disorder or condition is characterized by a defect in said patient'"'"'s cartilage.

19. The method of claim 18 wherein said disease, disorder or condition is chondrodysplasia and said gene encodes type II procollagen or type II collagen.

20. The method of claim 12 wherein said disease, disorder or condition is characterized by defect in said patient'"'"'s lungs.

21. The method of claim 20 wherein said disease, disorder or condition characterized is cystic fibrosis and said gene is a cystic fibrosis gene.

22. An implant device comprising:

a container having at least one membrane surface

stromal cells that comprise a gene construct, said gene construct comprising a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in said stromal cell.

23. The implant device of claim 22 wherein said membrane has a pore size of 0.3 microns.

24. The implant device of claim 22 having a membrane surface area of at least 100 mm².

25. The implant device of claim 22 comprising 10⁴ to 10¹¹ stromal cells.

26. The implant device of claim 22 comprising 10⁴ to 10⁸ stromal cells.

27. The implant device of claim 22 wherein said beneficial protein is selected form the group consisting of human growth hormone, obesity factor and human Factor VIII.

28. A method of treating an individual with a disease, disorder or condition which can be treated with a beneficial protein comprising the step of introducing into such an individual, immunologically isolated stromal cells that comprise a gene construct, said gene construct comprising a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in said stromal cell.

29. The method of claim 28 wherein said disease, disorder or condition which can be treated with a beneficial protein is a disease, disorder or conditions characterized by a gene defect.

30. The method of claim 29 wherein said beneficial protein is selected from the group consisting of human growth hormone and human Factor VIII.

31. The method of claim 28 wherein said immunologically isolated stromal cells are within an implant device that comprises said stromal cells and a container having at least one membrane surface.

32. The method of claim 31 wherein said membrane of said implant device has a pore size of 0.3 microns.

33. The method of claim 31 wherein said implant device has a membrane surface area of at least 100 mm².

34. The method of claim 31 wherein said implant device comprises 10⁴ to 10¹ stromal cells.

35. The method of claim 31 wherein said implant device comprises 10⁴ to 10⁸ stromal cells.

36. The method of claim 31 wherein said implant device is implanted into said individual subcutaneously.

37. Immunologically isolated stromal cells that comprise a gene construct, said gene construct comprising a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in said stromal cell.

38. The immunologically isolated stromal cells of claim 37 wherein said stromal cells are microencapsulated.

39. A method of treating patient who is suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect comprising the steps of:

a) obtaining a bone marrow sample from a donor who is not suffering from a disease, disorder or condition characterized by a bone or cartilage defect and who is syngeneic with said patient; and

,

b) administering a therapeutically effective amount of said bone marrow by intravenous infusion to said patient.

40. The method of claim 39 wherein said patent undergoes bone marrow ablation prior to administration of isolated stromal cells.

41. The method of claim 39 wherein said disease, disorder or condition is characterized by a defect in said patient'"'"'s bone.

42. The method of claim 41 wherein said disease, disorder or condition is osteogenesis imperfecta.

43. The method of claim 39 wherein said disease, disorder or condition is characterized by a defect in said patient'"'"'s cartilage.

44. The method of claim 43 wherein said disease, disorder or condition is chondrodysplasia.

45. A method of treating patient who suffering from a disease, disorder or condition characterized by a mutated, non-functioning or under-expressed gene which results in a defect in the bone, cartilage or lungs of said patient comprising the steps of:

a) obtaining a bone marrow sample from said patient;

b) isolating stromal cells from said sample;

c) culturing said stromal cells under conditions which result in replication of said stromal cells into an expanded culture of stromal cells; and

d) administering stromal cells of said expanded culture of stromal cells to said patient by intravenous infusion.

46. The method of claim 45 wherein said patent undergoes bone marrow ablation prior to administration of stromal cells.

47. The method of claim 46 wherein said stromal cells are administered by intravenous infusion to said patient together with hematopoietic precursor cells from said sample.

48. The method of claim 46 wherein said stromal cells are administered by intravenous infusion to said patient free from precursor cells from said sample.

49. The method of claim 45 wherein said disease, disorder or condition is characterized by a defect in said patient'"'"'s bone.

50. The method of claim 49 wherein said disease, disorder or condition is osteogenesis imperfecta or osteoporosis.

51. The method of claim 45 wherein said disease, disorder or condition is characterized by a defect in said patient'"'"'s cartilage.

52. The method of claim 46 wherein said disease, disorder or condition is chondrodysplasia or osteoarthritis.

53. The method of claim 45 wherein said disease, disorder or condition is characterized by defect in said patient'"'"'s lungs.

54. The method of claim 53 wherein said disease, disorder or condition characterized is cystic fibrosis and said gene is a cystic fibrosis gene.