Chemical approaches for generation of induced pluripotent stem cells

Patent metadata

Field	Value
Publication number	US-2016369244-A1
Application number	US-201615179657-A
Country	US
Kind code	A1
Filing date	Jun 10, 2016
Priority date	Mar 17, 2008
Publication date	Dec 22, 2016
Grant date	—

Abstract

The present invention provides for identification and use of small molecules to induce pluripotency in mammalian cells as well as other methods of inducing pluripotency.

First claim

What is claimed is:

1 . A method of producing an induced pluripotent stem cell from a mammalian non-pluripotent cell, the method comprising
(a) introducing into the mammalian non-pluripotent cell exogenous polynucleotides comprising an Oct4 encoding polynucleotide and a Klf4 encoding polynucleotide to obtain a reprogramming mammalian non-pluripotent cell; and (b) contacting the cell of step (a) with a small molecule composition comprising a GSK-3 inhibitor, thereby producing an induced pluripotent stem cell from the mammalian non-pluripotent cell.

2 . The method of claim 1 , wherein the small molecule composition further comprises a MEK inhibitor, an Erk inhibitor, or a TGFβ receptor/ALK5 inhibitor.

3 . The method of claim 2 , wherein the TGFβ receptor/ALK5 inhibitor is selected from the group consisting of: A-83-01 and SB431542.

4 . The method of claim 2 , wherein the MEK inhibitor is PD0325901.

5 . The method of claim 1 , wherein the GSK-3 inhibitor is selected from the group consisting of: CHIR99021, CHIR98014, and 6-bromoindirubin-3′-oxime (BIO).

6 . A composition comprising: a reprogramming mammalian non-pluripotent cell and a small molecule composition comprising a GSK-3 inhibitor.

7 . The composition of claim 6 , wherein the small molecule composition further comprises a MEK inhibitor, an Erk inhibitor, or a TGFβ receptor/ALK5 inhibitor.

8 . The composition of claim 7 , wherein the TGFβ receptor/ALK5 inhibitor is selected from the group consisting of: A-83-01 and SB431542.

9 . The composition of claim 7 , wherein the MEK inhibitor is PD0325901.

10 . The composition of claim 6 , wherein the GSK-3 inhibitor is selected from the group consisting of: CHIR99021, CHIR98014, and 6-bromoindirubin-3′-oxime (BIO).

11 . The composition of 6 , wherein the reprogramming mammalian non-pluripotent cell is contacted with exogenous polynucleotides comprising an Oct4 encoding polynucleotide and a Klf4 encoding polynucleotide to initiate reprogramming.

12 . A composition comprising: an induced pluripotent stem cell reprogrammed from a mammalian non-pluripotent cell, and a small molecule composition comprising a GSK-3 inhibitor.

13 . The composition of claim 12 , wherein the small molecule composition further comprises a MEK inhibitor, an Erk inhibitor, or a TGFβ receptor/ALK5 inhibitor.

14 . The composition of claim 13 , wherein the TGFβ receptor/ALK5 inhibitor is selected from the group consisting of: A-83-01 and SB431542.

15 . The composition of claim 13 , wherein the MEK inhibitor is PD0325901.

16 . The composition of claim 12 , wherein the GSK-3 inhibitor is selected from the group consisting of: CHIR99021, CHIR98014, and 6-bromoindirubin-3′-oxime (BIO).

17 . The composition of claim 13 , wherein the induced pluripotent stem cell is reprogrammed by introducing into mammalian non-pluripotent cell exogenous polynucleotides comprising an Oct4 encoding polynucleotide and a Klf4 encoding polynucleotide to initiate reprogramming.

Assignees

• Scripps Research Inst

Inventors

• Shi Yan
• Desponts Caroline
• Lin Tongxiang
• Li Wenlin
• Zhu Saiyong
• Ding Sheng

Classifications

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P25/28

  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 title

• A61P25/00

  Drugs for disorders of the nervous system · CPC title