Method of effecting de-differentiation of a cell

• US 9,102,920 B2
• Filed: 10/21/2008
• Issued: 08/11/2015
• Est. Priority Date: 05/06/2008
• Status: Expired due to Fees

• Current Assignee: Agency For Science Technology and Research (Government of Singapore)
• Original Assignee: Agency For Science Technology and Research (Government of Singapore)
• Inventors: Feng, Bo, Jiang, Jianming, Ng, Huck Hui, Lufkin, Thomas, Kraus, Petra
• Primary Examiner(s): Noble, Marcia S
• Application Number:
  US12/991,284
• Publication Number:
  US 20110165570A1
• Time in Patent Office: 2,485 Days
• US Class Current: 1/1
• CPC Class Codes: C12N 2501/38 with nuclear receptors
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 2501/606 c-Myc
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

The invention provides a method of effecting de-differentiation of an at least partially differentiated cell or of maintaining pluripotency and/or self-renewing characteristics of an undifferentiated cell. The method comprises increasing the amount or the activity of an Err protein, or a functional fragment thereof, in the cell.

Claims

1. A method of reprogramming a cell to produce a cell with pluripotency and self-renewing characteristics, the method comprising increasing the amount or activity of an estrogen-related (Err) protein, or a functional fragments thereof, Oct4, Sox2, and c-Myc in a cultured cell, wherein the cultured cell is a cultured somatic cell, a cultured precursor cell, or a cultured partially differentiated stem cell and wherein the Err protein is one of an Esrrb protein and Esrrg protein.

2. The method of claim 1, wherein the cultured partially differentiated stem cell is one of a lineage-restricted stem cell, a somatic stem cell, and a progenitor cell.

3. The method of claim 1, wherein increasing the amount of Err protein in the cultured cell comprises increasing the expression of the Err protein in the cultured cell.

4. The method of claim 3, wherein increasing the expression of the Err protein in the culture cell comprises increasing expression of a nucleic acid molecule encoding the Err protein in the cultured cell.

5. The method of claim 4, wherein the nucleic acid molecule is heterologous to the cultured cell.

6. The method of claim 4, wherein the nucleic acid molecule is RNA or DNA.

7. The method of claim 1, further comprising assessing one of:

(i) the amount or the activity of the Err protein, or a functional fragment thereof, in the cell, and

(ii) the presence of a marker of the differentiation status in the cell.

8. The method of claim 7, wherein the marker of the differentiation status in the cell is:

(i) the expression of one of Nanog, Oct4, Sox2, Sall4, Tc11, Tbx3, Eras, Klf2, Klf4, Klf5, Baf250a, BC031441, Eno3, Etv5, Gm1739, Gtf2h3, Hes6, Jub, Mtf2, Myod1, Nmyc1, Notch4, Nr5a2, Nrg2, Otx2, Rab2b, Rbpsuh, Rest, Stat3, Utf1, Tcfap2c, and Zfp553, or (ii) the methylation status of a promoter present in a gene encoding Nanog, Oct4, Sox2, Sall4, Tc11, Tbx3, Eras, Klf2, Klf4, Klf5, Baf250a, BC031441, Eno3, Etv5, Gm1739, Gtf2h3, Hes6, Jub, Mtf2, Myod1, Nmyc1, Notch4, Nr5a2, Nrg2, Otx2, Rab2b, Rbpsuh, Rest, Stat3, Utf1, Tcfap2c, or Zfp553.

9. The method of claim 7, wherein assessing the amount and/or activity of the Err protein comprises assessing the amount and/or activity over a period of time.

10. The method of claim 7, wherein assessing the amount of the Err protein in the cell is achieved by measuring gene expression of the Err protein in the cell.

11. The method of claim 10, further comprising comparing the result of the measurement of Err gene expression with that of a control measurement.

12. The method of claim 11, wherein the control measurement comprises a measurement wherein Err gene expression is not modulated.

13. The method of claim 7, wherein assessing the presence of a marker of the differentiation status in the cell comprises measuring the expression and/or the activity of the marker over a period of time.

14. The method of claim 1, wherein the cell is obtained from an animal.

15. The method of claim 14, wherein the animal is one of a fish, an amphibian, a bird and a mammal.

16. The method of claim 15, wherein the cell is a mammalian somatic cell selected from the group consisting of a fibroblast, a myeloid cell, a B lymphocyte, a T lymphocyte, a dendritic cell, a keratinocyte, an adipose cell, a mesenchymal cell, an epithelial cell, an epidermal cell, a chondrocyte, a hepatocyte, a cumulus cell, a neural cell, a glial cell, an astrocyte, a cardiac cell, an esophageal cell, a muscle cell, a pancreatic beta cell, a melanocyte, a hematopoietic cell, a macrophage, a monocyte, and a mononuclear cell.

17. The method of claim 1, wherein increasing the amount or activity of the Err protein in the cultured cell comprises contacting the cultured cell with a compound that increases the expression or activity of the Err protein.