Reprogramming of cells to a new fate

• US 10,308,912 B2
• Filed: 12/12/2016
• Issued: 06/04/2019
• 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
• Application Number:
  US15/376,536
• Publication Number:
  US 20170159019A1
• Time in Patent Office: 904 Days
• Field of Search: None
• 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 pharmaceutical composition comprising an isolated population of cells having a second non-pluripotent cell fate, wherein the cells are obtained by a method of converting animal cells from a first non-pluripotent cell fate, and wherein the method comprises:

(a) introducing a polynucleotide encoding an Oct4 polypeptide, and optionally one or more reprogramming factors comprising polynucleotides encoding a Klf polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide, into cells having a first non-pluripotent cell fate, or contacting cells having a first non-pluripotent cell fate with an Oct4 polypeptide, and optionally one or more reprogramming factors comprising a Klf 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 cells from step (b) under conditions to generate the population of cells having the second non-pluripotent cell fate, wherein the cells having the first non-pluripotent cell fate are mesodermal, ectodermal or endodermal cells, and wherein the cells having the second non-pluripotent cell fate from step (c) are in the same or different cell lineage as the cells having the first non-pluripotent cell fate.

2. The pharmaceutical composition of claim 1, further comprising a physiologically acceptable carrier.

3. The pharmaceutical composition of claim 1, further comprising one or more of:

a preservative, an antimicrobial agent, an anti-oxidant, a chelating agent, and an inert gas.

4. The pharmaceutical composition of claim 1, wherein the cells having the first non-pluripotent cell fate are fibroblasts or neural precursor cells.

5. The pharmaceutical composition of claim 1, wherein the cells having the second non-pluripotent cell fate are cells of mesodermal lineage, cells of ectodermal lineage, or cells of endodermal lineage.

6. The pharmaceutical composition of claim 1, wherein the cells having the second non-pluripotent cell fate are cardiac cells, neural cells, induced definitive endoderm (iDE) cells, pancreatic cells, hematopoietic cells, or retinal pigmented epithelium cells.

7. A method of treating a subject by administering the pharmaceutical composition of claim 1 to a subject in need of treatment.

8. The method of claim 7, wherein the cells having the second non-pluripotent cell fate comprised in the pharmaceutical composition are allogenic or autogenic to the subject.

9. The method of claim 7, wherein the subject is in need of heart tissue transplantation, or has a congenital heart defect, ischemic heart tissue, cardiomyopathy, diabetes, nerve damage, macular degeneration, photoreceptor dystrophy, merTK dystrophy, autoimmune disorder, a hematological malignancy, a solid tumor, cancer, or an infection associated with HIV, RSV, EBV, CMV, adenovirus, or BK polyomavirus.

10. A method of manufacturing a population of cells having a second non-pluripotent cell fate obtained by a method of converting animal cells having a first non-pluripotent cell fate, wherein the method comprises:

(a) (i) introducing factors comprising polynucleotides encoding an Oct4 polypeptide and encoding one or more of a Klf polypeptide, a Sox2 polypeptide, or a c-Myc polypeptide, into cells having a first non-pluripotent cell fate, or (ii) contacting cells having a first non-pluripotent cell fate with an Oct4 polypeptide and one or more reprogramming factors comprising a Klf 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); and

(c) inducing differentiation of the cells from step (b) under conditions to generate the population of cells having the second non-pluripotent cell fate, wherein the cells having the first non-pluripotent cell fate are mesodermal, ectodermal or endodermal cells, and wherein the cells having the second non-pluripotent cell fate from step (c) are in the same or different cell lineage as the cells having the first non-pluripotent cell fate.

11. The method of claim 10, wherein the cells having the first non-pluripotent cell fate are fibroblasts or neural precursor cells.

12. The method of claim 10, wherein the cells having the second non-pluripotent cell fate are cells of mesodermal lineage, cells of ectodermal lineage, or cells of endodermal lineage.

13. The method of claim 10, wherein the cells having the second non-pluripotent cell fate are cardiac cells, neural cells, induced definitive endoderm (iDE) cells, pancreatic cells, hematopoietic cells, or retinal pigmented epithelium cells.