Chemical approaches for generation of induced pluripotent stem cells

• US 9,771,563 B2
• Filed: 06/10/2016
• Issued: 09/26/2017
• Est. Priority Date: 03/17/2008
• Status: Active Grant

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Shi, Yan, Desponts, Caroline, Lin, Tongxiang, Li, Wenlin, Zhu, Saiyong, Ding, Sheng
• Primary Examiner(s): Noble, Marcia S
• Application Number:
  US15/179,657
• Publication Number:
  US 20160369244A1
• Time in Patent Office: 473 Days
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 11/08 Bronchodilators
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• 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 15/85 for animal cells
• CPC Class Codes: C12N 2500/14 Calcium; Ca chelators; Calcitonin
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/065 Modulators of histone acetylation
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 2501/604 : Klf-4
• CPC Class Codes: C12N 2501/605 : Nanog
• CPC Class Codes: C12N 2501/608 : Lin28
• CPC Class Codes: C12N 2501/727 : Kinases (EC 2.7.)
• CPC Class Codes: C12N 2506/08 : from cells of the nervous system
• CPC Class Codes: C12N 2506/094 : from keratinocytes
• CPC Class Codes: C12N 2506/1307 : from adult fibroblasts
• CPC Class Codes: C12N 2506/28 : from vascular endothelial cells
• 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 present invention provides for identification and use of small molecules to induce pluripotency in mammalian cells as well as other methods of inducing pluripotency.

Claims

1. A composition comprising:

(i) a mammalian non-pluripotent cell comprising exogenous polynucleotides comprising an Oct4 encoding polynucleotide and a Klf encoding polynucleotide; and

(ii) a small molecule composition comprising a GSK-3 inhibitor.

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

receptor/ALK5 inhibitor.

3. The composition of claim 2, wherein the TGFβ

receptor/ALK5 inhibitor is selected from the group consisting of;

A-83-01 and SB431542.

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

5. The composition of claim 1, wherein the GSK-3 inhibitor is selected from the group consisting of:

CHIR99021, CHIR98014, and 6-bromoindirubin-3′

-oxime (BIO).

6. The composition of 1, wherein the mammalian non-pluripotent cell is contacted with exogenous polynucleotides comprising an Oct4 encoding polynucleotide and a Klf4 encoding polynucleotide to initiate reprogramming.

7. 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.

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

receptor/ALK5 inhibitor.

9. The composition of claim 8, wherein the TGFβ

receptor/ALK5 inhibitor is selected from the group consisting of;

A-83-01 and SB431542.

10. The composition of claim 8, wherein the MEK inhibitor is PD0325901.

11. The composition of claim 7, wherein the GSK-3 inhibitor is selected from the group consisting of:

CHIR99021, CHIR98014, and 6-bromoindirubin-3′

-oxime (BIO).

12. The composition of claim 8, wherein the induced pluripotent stem cell is reprogrammed by introducing into mammalian non-pluripotent cell exogenous polynucleotides comprising an Oct4 encoding polynucleotide and a Klf encoding polynucleotide to initiate reprogramming.