KIDNEY-DERIVED CELLS AND METHODS OF USE IN TISSUE REPAIR AND REGENERATION

• US 20080112939A1
• Filed: 10/11/2007
• Published: 05/15/2008
• Est. Priority Date: 10/12/2006
• Status: Active Grant

• Current Assignee: Depuy Synthes Products Incorporated (Johnson & Johnson)
• Original Assignee: Ethicon Incorporated (Johnson & Johnson)
• Inventors: Seyda, Agnieszka, Gosiewska, Anna, Colter, David C., Buensuceso, Charito S.
• Granted Patent: US 10,500,230 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• 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 35/22 Urine; Urinary tract, e.g. kidney or bladder; Intraglomerular mesangial cells; Renal mesenchymal cells; Adrenal gland
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• 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 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 5/0686 Kidney cells
• CPC Class Codes: C12N 5/0687 Renal stem cells; Renal progenitors

Abstract

Isolated or purified mammalian kidney-derived cell populations from mammalian kidney tissue are provided. Methods are provided for the isolation and purification of the mammalian kidney-derived cell population. Methods for treating kidney disease are provided by administration of the isolated or purified mammalian kidney-derived cell population to a mammalian subject.

Claims

1. An isolated or purified mammalian kidney-derived cell population, said cell population capable of self-renewal and expansion in culture, wherein the cell population is positive for expression of at least one of Oct-4, Rex-1, Pax-2, Cadherin-11, FoxD1, WT1, Eya1, HNF3B, CXC-R4, Sox-17, EpoR, BMP2, BMP7, or GDF5 and negative for the expression of at least one of Sox2, FGF4, hTert, Wnt-4, SIX2 or GATA-4.

2. The cell population of claim 1 that is positive for expression of at least one of Eya1, WT1, FoxD1, BMP7, BMP2, GDF5, EpoR or Rex-1, and negative for expression of at least one of Sox2, FGF4, hTert or Wnt-4.

3. The cell population of claim 1 that is positive for at least one of cell-surface markers HLA I, CD24, CD29, CD44, CD49c, CD73, CD166, or SSEA-4, and negative for at least one of cell-surface markers HLA II, CD31, CD34, CD45, CD56, CD80, CD86, CD104, CD105, CD117, CD133, CD138, CD141, or E-cadherin.

4. The cell population of claim 1 that is positive for at least one of the surface markers HLA I, CD166 or SSEA-4, and negative for at least one of the cell-surface markers HLA II, CD80, CD86, CD133, CD141 or E-cadherin.

5. The cell population of claim 1 that is positive for the cell-surface marker HLA I, and negative for at least one of cell-surface markers HLA II, CD80, or CD86.

6. The cell population of claim 5 wherein the cell population is non-immunogenic for allogeneic transplantation in a mammalian subject.

7. The cell population of claim 1, wherein the cells are derived from kidney cortex.

8. The cell population of claim 1, wherein the cells are derived from kidney medulla.

9. The cell population of claim 1, wherein the cells are derived from kidney subcapsular region.

10. The cell population of claim 1, wherein the cell population secretes at least one of trophic factors FGF2, HGF, TGFα

, TIMP-1, TIMP-2, MMP-2 or VEGF and does not secrete at least one of trophic factors PDGF-bb or IL12p70.

11. The cell population of claim 1, wherein the cell population comprises human cells, primate cells, or rodent cells.

12. The cell population of claim 1, wherein the cell population comprises kidney progenitor cells.

13. The cell population of claim 12, wherein the kidney progenitor cells are capable of differentiating to adipocytes or osteoblasts.

14. A method for treating ischemic kidney disease in a mammalian subject comprising, administering to the mammalian subject a therapeutically effective amount of the cell population according to any one of claims 1 to 13, thereby reducing or eliminating the ischemic kidney tissue disease in the mammalian subject.

15. The method of claim 14, wherein the administration of said cell population induces an effect selected from:

(a) formation of blood vessels supplying blood to the ischemic tissue;

(b) blood flow to the ischemic tissue;

(c) oxygen supply to the ischemic tissue; and

(d) combinations thereof.

16. The method of claim 14, wherein the administration of said cell population induces formation of kidney subcapsular region tissue, kidney cortex tissue, or kidney medulla tissue.

17. The method of claim 14, wherein the mammalian subject is a human, a primate, or a rodent.

18. A method for replacing renal tissues, organs, components or structures which are damaged due to trauma, age, metabolic or toxic injury, disease or idiopathic loss in a mammalian subject comprising, administering to the mammalian subject a therapeutically effective amount of the cell population according to any one of claims 1 to 13, thereby reducing or eliminating the damaged tissues or organs and restoring kidney function in the mammalian subject.

19. The method of claim 18, wherein administration of said cell population induces an effect selected from:

(a) formation of blood vessels supplying blood to the kidney tissue;

(b) blood flow to the kidney tissue;

(c) oxygen supply to the kidney tissue; and

(d) combinations thereof.

20. The method of claim 18, wherein administration of said cell population induces formation of kidney subcapsular region tissue, kidney cortex tissue, or kidney medulla tissue.

21. The method of claim 18, wherein the mammalian subject is a human, a primate, or a rodent.

22. A method for preparing an isolated mammalian kidney derived cell population comprising,

obtaining tissue from a subcapsular region, a cortex or a medulla of a mammalian kidney,

incubating the tissue in the presence of one or more metalloprotease, neutral protease, or mucolytic enzyme to disassociate at least a portion of the tissue into individual cells,

plating the cells on a substrate,

identifying within said cells a subset of cells that is positive for expression of at least one of Oct-4, Rex-1, Pax-2, Cadherin-11, FoxD1, WT1, Eya1, HNF3B, CXC-R4, Sox-17, EpoR, BMP2, BMP7, or GDF5 and negative for the expression of at least one of Sox2, FGF4, hTert, Wnt-4, SIX2 or GATA-4, and

isolating said subset of cells to provide said mammalian kidney-derived cell population.

23. The method of claim 22, wherein the cell population is positive for expression of at least one of Eya1, WT1, FoxD1, BMP7, BMP2, GDF5, EpoR or Rex-1, and negative for expression of at least one of Sox2, FGF4, hTert or Wnt-4.

24. The method of claim 22, further comprising the step of identifying a subset of cells that are positive for the cell-surface marker HLA I, and negative for at least one of the cell-surface markers HLA II, CD80, or CD86.

25. The method of claim 24, wherein the cell population is non-immunogenic for allogeneic transplantation in the mammalian subject.

26. The method of claim 22, wherein said tissue is from the cortex of a mammalian kidney.

27. The method of claim 22, wherein said tissue is from the medulla of a mammalian kidney.

28. The method of claim 22, wherein said tissue is from the subcapsular region of a mammalian kidney.

29. The method of claim 22, wherein the substrate is a tissue culture flask, a scaffold, or a hollow, fiber-based device.

30. The method of claim 22, further comprising isolating a single mammalian kidney-derived cell by limiting dilution and expanding the single cell in cell culture.

31. The method of claim 22, wherein said tissue is from a human kidney.

32. A method for treating a disease in a mammalian subject with gene therapy comprising,

obtaining an isolated or purified population of mammalian kidney-derived cells capable of self-renewal and expansion in culture, wherein the cell population is positive for expression of at least one of Oct-4, Rex-1, Pax-2, Cadherin-11, FoxD1, WT1, Eya1, HNF3B, CXC-R4, Sox-17, EpoR, BMP2, BMP7, or GDF5 and negative for the expression of at least one of Sox2, FGF4, hTert, Wnt-4, SIX2 or GATA-4,

genetically altering at least one cell of the isolated or purified mammalian kidney-derived cell population to produce a therapeutic gene product,

expanding the genetically altered cells in culture, and

administering the genetically altered cells to the mammalian subject to reduce or eliminate the disease in the mammalian subject.

33. The method of claim 32, wherein the cell population is positive for expression of at least one of Eya1, WT1, FoxD1, BMP7, BMP2, GDF5, EpoR or Rex-1, and negative for expression of at least one of Sox2, FGF4, hTert or Wnt-4.

34. The method of claim 32, wherein the cell population is positive for the cell-surface marker HLA I, and negative for at least one of the cell-surface markers HLA II, CD80, or CD86.

35. The method of claim 34, wherein the cell population is non-immunogenic for allogeneic transplantation in the mammalian subject.

36. The method of claim 32, wherein the genetically altered cells are administered to the kidney of the mammalian subject to treat a kidney disease, kidney injury, kidney ischemia, or genetic defect in the mammalian subject.

37. The method of claim 32, wherein the mammalian subject is a human, a primate, or a rodent.

38. The method of claim 32, wherein the mammalian kidney-derived cell population is derived from the kidney of a human, a primate, or a rodent.

39. The method of claim 38, wherein the mammalian kidney-derived cell population are derived from the kidney of a human and the cells are transplanted in utero to produce human kidney cells in the prenatal or postnatal human subject following transplantation, wherein the cells produce therapeutic products in the human or animal to reduce or eliminate the disease in the human subject.

40. A method for screening a drug candidate to treat a disorder involving renal cells in a mammalian subject comprising the steps of:

(a) providing an isolated or purified mammalian kidney-derived cell population, said population capable of self-renewal and expansion in culture, wherein the cell population is positive for expression of at least one of Oct-4, Rex-1, Pax-2, Cadherin-11, FoxD1, WT1, Eya1, HNF3B, CXC-R4, Sox-17, EpoR, BMP2, BMP7, or GDF5 and negative for the expression of at least one of Sox2, FGF4, hTert, Wnt-4, SIX2 or GATA-4;

(b) culturing the mammalian kidney-derived cell population under proliferation conditions to obtain a cellular composition comprising cells with potential or increased potential for self-renewal and expansion in vitro;

(c) exposing the cultured cells obtained from step (a) or (b) to the drug candidate; and

(d) detecting the presence or absence of an effect of the drug candidate on the survival of the cells or on a morphological, functional, or physiological characteristic and/or molecular biological property of said cells, whereby an effect altering cell survival, a morphological, functional or physiological characteristic of the cells, and/or a molecular biological property of the cells indicates an activity of the drug candidate to treat the renal cell disorder.

41. The method of claim 40, wherein the cell population is positive for expression of at least one of Eya1, WT1, FoxD1, BMP7, BMP2, GDF5, EpoR or Rex-1, and negative for expression of at least one of Sox2, FGF4, hTert or Wnt-4.

42. The method of claim 40, wherein the morphological, functional, or physiological characteristic of the cell is increased formation of renal tubules or expression of BMP or BMP receptor.

43. The method of claim 40, wherein the morphological, functional, or physiological characteristic of the cell is increased secretion of at least one or more of the trophic factors FGF2, HGF, TGFα

, TIMP-1, TIMP-2, VEGF, or MMP-2.

44. The method of claim 40, wherein the disorder is ischemic kidney disease or damage to kidney tissues, organs, components or structures due to trauma, age, metabolic or toxic injury, disease or idiopathic loss in a mammalian subject.

45. A method for assaying toxicity of a test substance to affect kidney cell function comprising the steps of:

(a) providing an isolated or purified mammalian kidney-derived cell population capable of self-renewal and expansion in culture, wherein the cell population is positive for expression of at least one of Oct-4, Rex-1, Pax-2, Cadherin-11, FoxD1, WT1, Eya1, HNF3B, CXC-R4, Sox-17, EpoR, BMP2, BMP7, or GDF5 and negative for the expression of at least one of Sox2, FGF4, hTert, Wnt-4, SIX2 or GATA-4;

(b) exposing the cell population to the test substance; and

(c) detecting the presence or absence of an effect of the test substance on the survival of the cells in the population or on a morphological, functional, or physiological characteristic and/or molecular biological property of cells in the population, whereby an effect altering cell survival, a morphological, functional, or physiological characteristic of the cells and/or a molecular biological property of cells in the population indicates the toxicity of the test substance to kidney cell function.

46. The method of claim 45, wherein the cell population is positive for expression of at least one of Eya1, WT1, FoxD1, BMP7, BMP2, GDF5, EpoR or Rex-1, and negative for expression of at least one of Sox2, FGF4, hTert or Wnt-4.

47. The method of claim 45, wherein the morphological, functional, or physiological characteristic of the cells is formation of renal tubules or expression of BMP or BMP receptor.

48. The method of claim 47, wherein increased toxicity of the test substance is measured as decreased formation of renal tubules or decreased expression of BMP or BMP receptor.

49. The method of claim 45, wherein the morphological, functional, or physiological characteristic of the cells is secretion of at least one or more of trophic factors FGF2, HGF, TGFα

, TIMP-1, TIMP-2, VEGF, or MMP-2.