Conversion of liver stem and progenitor cells to pancreatic functional cells

• US 20050053588A1
• Filed: 10/18/2002
• Published: 03/10/2005
• Est. Priority Date: 10/18/2001
• Status: Abandoned Application

• Current Assignee: Ixion Biotechnology Inc. (Galderma Holding SA)
• Original Assignee: Ixion Biotechnology Inc. (Galderma Holding SA)
• Inventors: Yin, Li
• Application Number:
  US10/493,536
• Publication Number:
  US 20050053588A1
• 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: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/117 Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/13 Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
• CPC Class Codes: C12N 2501/148 Transforming growth factor alpha [TGF-a]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/24 Interferons [IFN]
• CPC Class Codes: C12N 2501/335 Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 2501/345 Gastrin; Cholecystokinins [CCK]
• 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 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/998 : Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/03 : from non-embryonic pluripotent stem cells
• CPC Class Codes: C12N 2506/14 : from hepatocytes
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 2510/02 : Cells for production
• CPC Class Codes: C12N 5/0676 : Pancreatic cells

Abstract

The subject invention a method for converting liver stem/progenitor cells to a pancreatic functional cell by transfecting said liver cells with a pancreatic development gene and/or by culturing with pancreatic differentiation factors. The resulting cells produce and secrete insulin protein in response to glucose stimulation.

Claims

1. A method of converting a liver stem/progenitor cell to a pancreatic functional cell, wherein said pancreatic functional cell produces and secretes insulin in response to glucose stimulation, said method comprising:

transfecting said liver stem/progenitor cell with a pancreatic development gene, culturing said liver stem/progenitor cell in a medium comprising factors that induce differentiation into the pancreatic functional cell, or both, whereby said transfected cell is converted to the pancreatic functional cell.

2. The method of claim 1, wherein the pancreatic functional cell is a cell of the pancreatic endocrine lineage.

3. The method of claim 2, wherein said converted cell is selected from the group consisting of islet producing stem cells (IPSCs), islet progenitor cells (IPCs) and islet-like structures or IPC-derived islets (IdIs).

4. The method of claim 1, wherein the pancreatic functional cell further expresses a marker of the liver stem/progenitor cell.

5. The method of claim 1, wherein said liver stem/progenitor cell is selected from the group consisting of hepatoblasts and liver oval cells.

6. The method of claim 1, wherein said liver stem/progenitor cell expresses at least one marker selected from the group consisting of hematopoietic markers and liver oval or hepatoblast cell markers.

7. The method of claim 6, wherein said hematopoietic markers are selected from the group consisting of CD34, Thy1.1 and CD45.

8. The method of claim 6, wherein said liver hepatoblast or oval cell markers are selected from the group consisting of α

-fetal protein, albumin, cytokeratin 14 (CK14), c-kit, OC.2, OC.3, OC.10, OV1 and OV6.

9. The method of claim 1, wherein said pancreatic development gene is selected from the group consisting of Pdx1, Hlxb9, Isl1, ngn3, Nkx2.2 Pax6, NeuroD/β

2, Nkx6.1 and Pax4.

10. The method of claim 9, wherein said pancreatic development gene is Pdx-1.

11. The method of claim 1, wherein said factors are selected from the group consisting of dexamethasone, glucagon-like peptide-1 (GLP-1), exendin-4, gastrin, interferon-γ

(IFNγ

), hepatocyte growth factor (HGF), epidermal growth factor (EGF), β

-cellulin, activin-A, keratinocyte growth factor (KGF), fibroblast growth factor (FGF), transforming growth factor-α

(TGF-α

), transforming growth factor-β

(TGF-β

), nerve growth factor (NGF), insulin-like growth factors (IGFs), islet neogenesis associated protein (INGAP), vascular endothelial growth factor (VEGF), nicotinamide, retinoic acid, sodium butyrate and any combination thereof.

12. The method of claim 1, wherein said converted cell expresses a pancreatic message selected from the group consisting of insulin I (InsI), insulin II (InsII), glucagon, somatostatin, pancreatic polypeptide (PP), amylase, elastase, glucose transporter 2 (GLUT2), glucokinase, PC1, PC2, PC3, carboxypeptidase E (CPE), Pdx1, Hlxb9, Isl1, ngn3, Nkx2.2, Pax6, NeuroD/β

2, Nkx6.1 and Pax4.

13. The method of claim 1, wherein said converted cell expresses a pancreatic protein selected from the group consisting of InsI, InsII, glucagon, somatostatin, PP, amylase, elastase, GLUT2, glucokinase, PC1, PC2, PC3, CPE, Pdx1, Hlxb9, Isl1, ngn3, Nkx2.2, Pax6, NeuroD/β

2, Nkx6.1 and Pax4.

14. A pancreatic functional cell produced by the method of claim 1.

15. The pancreatic functional cell of claim 14, wherein the cell further expresses Pdx1, amylase and insulin II.

16. A method for producing an endocrine hormone comprising converting liver stem/progenitor cells according to the method of claim 1, and further comprising:

culturing said converted cells; and

recovering endocrine hormone from said cell culture.

17. A liver stem/progenitor cell that has been transfected with a pancreatic development gene.

18. The method of claim 17, wherein said liver stem/progenitor cell is selected from the group consisting of hepatoblasts and liver oval cells.

19. The liver stem/progenitor cell of claim 17 wherein the pancreatic development gene is selected from the group consisting of Pdx1, Hlxb9, Isl1, ngn3, Nkx2.2, Pax6, NeuroD/β

2, Nkx6.1 and Pax4.

20. The liver stem/progenitor cell of claim 19 wherein the pancreatic development gene is Pdx1.

21. A liver stem/progenitor cell that has been cultured in a medium comprising factors that induce differentiation into a pancreatic functional cell.

22. The liver stem/progenitor cell of claim 21 wherein said factors are selected from the group consisting of dexamethasone, glucagon-like peptide-1 (GLP-1), exendin4, gastrin, interferon-γ

(IFNγ

), hepatocyte growth factor (HGF), epidermal growth factor (EGF), β

-cellulin, activin-A, keratinocyte growth factor (KGF), fibroblast growth factor (FGF), transforming growth factor-α

(TGF-α

), transforming growth factor-β

(TGF-β

), nerve growth factor (NGF), insulin-like growth factors (IGFs), islet neogenesis associated protein (INGAP), vascular endothelial growth factor (VEGF), nicotinamide, retinoic acid, sodium butyrate and any combination thereof.