Reprogramming of cells to a new fate

• US 9,556,417 B2
• Filed: 03/17/2016
• Issued: 01/31/2017
• Est. Priority Date: 06/14/2010
• Status: Active Grant

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Efe, Jem A., Kim, Janghwan, Zhu, Saiyong, Hilcove, Simon, Ding, Sheng
• Primary Examiner(s): Weidner, Adam M
• Application Number:
  US15/072,769
• Publication Number:
  US 20160264938A1
• Time in Patent Office: 320 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 35/30 Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
• CPC Class Codes: A61K 35/33 Fibroblasts
• CPC Class Codes: A61K 35/34 Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes vascular smooth muscle A61K35/44
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• 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 2501/606 c-Myc
• CPC Class Codes: C12N 2501/727 Kinases (EC 2.7.)
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/08 from cells of the nervous system
• CPC Class Codes: C12N 2506/1307 from adult fibroblasts
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0602 Vertebrate cells
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0607 : Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0618 : Cells of the nervous system
• CPC Class Codes: C12N 5/0619 : Neurons
• CPC Class Codes: C12N 5/0623 : Stem cells
• CPC Class Codes: C12N 5/0657 : Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0678 : Stem cells; Progenitor cells; Precursor cells

Abstract

The present invention generally provides methods and compositions for transdifferentiation of an animal cell from a first non-pluripotent cell fate to a second non-pluripotent cell fate. Also provided are methods and compositions for the transdifferentiation of an animal cell from a non-pluripotent mesodermal, endodermal, or ectodermal cell fate to a different non-pluripotent mesodermal, endodermal, or ectodermal cell fate.

Claims

1. A method of converting an animal cell from a first non-pluripotent cell fate to a second non-pluripotent cell fate, the method comprising:

(a) introducing a polynucleotide encoding an Oct4 polypeptide, and optionally one or more reprogramming factors comprising polynucleotides encoding a Klf4 polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide, into a first non-pluripotent cell, or contacting a first non-pluripotent cell with an Oct4 polypeptide, and optionally one or more reprogramming factors comprising a Klf4 polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide,

(b) limiting the expression of endogenous Nanog in the cells from step (a) to a level substantially lower than the level of expression of endogenous Nanog in an induced pluripotent cell (iPSC), thereby generating a non-pluripotent intermediate cell and

(c) inducing differentiation of the cell under conditions to generate a cell having a second non-pluripotent cell fate,

wherein the first non-pluripotent cell is a mesodermal, ectodermal or endodermal cell, and wherein the cell having the second non-pluripotent cell fate from step (c) is in the same or different cell lineage as the first non-pluripotent cell.

2. The method of claim 1, wherein step (b) comprises eliminating exogenous LIF.

3. The method of claim 1, wherein the first non-pluripotent cell is a fibroblast or a neural precursor cell.

4. The method of claim 1, wherein inducing differentiation of the non-pluripotent intermediate cell to the cell having the second non-pluripotent cell fate comprises contacting the non-pluripotent intermediate cell with BMP4, a calcium channel agonist, a Gas activating agent, a cAMP analog, and/or a GSK-3 inhibitor.

5. The method of claim 1, wherein the method is conducted in vitro.