METHODS OF DIFFERENTIATING AND PROTECTING CELLS BY MODULATING THE P38/MEF2 PATHWAY

• US 20110212070A1
• Filed: 03/31/2011
• Published: 09/01/2011
• Est. Priority Date: 06/05/2000
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Sanford-Burnham Medical Research Institute (Sanford Burnham Prebys Medical Discovery Institute)
• Inventors: Lipton, Stuart A., Okamoto, Shu-ichi
• Granted Patent: US 8,236,563 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: A61P 25/00 Drugs for disorders of the nervous system
• 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: C12N 2501/385 of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2503/00 Use of cells in diagnostics
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0619 Neurons
• CPC Class Codes: G01N 33/6887 from muscle, cartilage or connective tissue

Abstract

The present invention provides a method of differentiating progenitor cells to produce a population containing protected neuronal cells. A method of the invention includes the steps of contacting the progenitor cells with a differentiating agent; and introducing into the progenitor cells a nucleic acid molecule encoding a MEF2 polypeptide or an active fragment thereof, thereby differentiating the progenitor cells to produce a population containing protected neuronal cells. In one embodiment, the MEF2 polypeptide is human MEF2C or an active fragment thereof.

Claims

1. A method of differentiating progenitor cells, comprising the steps of:

(a) contacting said progenitor cells with a differentiating agent; and

(b) introducing into said progenitor cells a nucleic acid molecule encoding a MEF2 polypeptide or an active fragment thereof,

thereby differentiating said progenitor cells to produce a cell population containing protected neuronal cells.

2. The method of claim 1, wherein said MEF2 polypeptide is human MEF2C, or an active fragment thereof.

3. The method of claim 1, wherein said MEF2 polypeptide is constitutively active.

4. The method of claim 3, wherein said constitutively active MEF2 polypeptide is a MEF2/VP16 fusion protein.

5. The method of claim 3, wherein said constitutively active MEF2 polypeptide contains one or more serine/threonine to aspartic acid/glutamic acid substitutions in the MEF2 transactivation domain.

6. The method of claim 3, further comprising inhibiting caspase activity in said progenitor cells.

7. The method of claim 1, wherein said progenitor cells are human stem cells.

8. The method of claim 1, wherein said progenitor cells are embryonic stem cells.

9. The method of claim 8, wherein said embryonic stem cells are human embryonic stem cells.

10. The method of claim 1, wherein said progenitor cells are hematopoietic progenitor cells.

11. The method of claim 10, wherein said hematopoietic progenitor cells are human hematopoietic progenitor cells.

12. The method of claim 1, further comprising selecting CD133-positive human progenitor cells.

13. The method of claim 1, further comprising selecting CD133-positive/CD34-positive human progenitor cells.

14. The method of claim 1, further comprising selecting CD133-positive/CD34-negative human progenitor cells.

15. The method of claim 1, further comprising selecting CD133-positive/CD34-negative/CD45-negative human progenitor cells.

16. The method of claim 1, further comprising selecting CD34-negative/CD38-negative/Lin-negative human progenitor cells.

17. The method of claim 1, further comprising selecting CD34-positive/CD38-negative/Lin-negative/Thy-1-negative human progenitor cells.

18. The method of claim 1, wherein said differentiating agent is retinoic acid.

19. The method of claim 1, wherein said differentiating agent is selected from the group consisting of neurotrophic factor 3, epidermal growth factor, insulin-like growth factor 1 and a platelet-derived growth factor.

20. The method of claim 1, wherein said population containing protected neuronal cells comprises at least 50% neuronal cells.

21. The method of claim 1, further comprising the step of

(c) transplanting cells comprising a nucleic acid molecule encoding a MEF2 polypeptide or an active fragment thereof into a patient to produce a cell population containing protected neuronal cells in said patient.

22-57. -57. (canceled)