# US 10,308,912 B2 · Reprogramming of cells to a new fate

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,308,912 B2
- Document ID: US10308912B2
- Status: Granted
- Kind: B2
- Application number: 15/376,536
- Filed: 2016-12-12
- Published: 2017-06-08
- Granted: 2019-06-04
- Earliest claimed: 2010-06-14
- Assignees: The Scripps Research Institute
- Applicants: The Scripps Research Institute
- Inventors: Jem A. Efe; Janghwan Kim; Saiyong Zhu; Simon Hilcove; Sheng Ding
- Web page: https://patentgrep.com/patent/US10308912B2

## CPC

- C12N 5/0676: {Pancreatic cells}
- 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
- A61K 35/33: Fibroblasts
- A61K 35/34: Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes (vascular smooth muscle A61K35/44)
- A61K 35/39: Pancreas; Islets of Langerhans (Langerhans cells of epidermis A61K35/36)
- C12N 5/0602: {Vertebrate cells}
- C12N 5/0603: {Embryonic cells (production of embryos, nuclear transfer A01K67/027); Embryoid bodies}
- C12N 5/0607: {Non-embryonic pluripotent stem cells, e.g. MASC (induced pluripotent stem cells C12N5/0696)}
- C12N 5/0618: {Cells of the nervous system}
- C12N 5/0619: {Neurons}
- C12N 5/0623: {Stem cells}
- C12N 5/0657: {Cardiomyocytes; Heart cells}
- C12N 5/0678: {Stem cells; Progenitor cells; Precursor cells}
- C12N 15/85: for animal cells
- C12N 2501/01: Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
- C12N 2501/155: Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
- C12N 2501/235: Leukemia inhibitory factor [LIF]
- C12N 2501/602: Sox-2
- C12N 2501/603: Oct-3/4
- C12N 2501/604: Klf-4
- C12N 2501/605: Nanog
- C12N 2501/606: c-Myc
- C12N 2501/727: Kinases (EC 2.7.)
- C12N 2506/02: from embryonic cells
- C12N 2506/08: from cells of the nervous system
- C12N 2506/1307: from adult fibroblasts
- C12N 2510/00: Genetically modified cells
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## 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

### Claim 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.

### Claim 2 (depends on 1)

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

### Claim 3 (depends on 1)

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.

### Claim 4 (depends on 1)

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

### Claim 5 (depends on 1)

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.

### Claim 6 (depends on 1)

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.

### Claim 7 (depends on 1)

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

### Claim 8 (depends on 7)

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.

### Claim 9 (depends on 7)

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.

### Claim 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.

### Claim 11 (depends on 10)

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

### Claim 12 (depends on 10)

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.

### Claim 13 (depends on 10)

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.