# US 12,006,516 B2 · Differentiation of pancreatic endocrine cells

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- Number: US 12,006,516 B2
- Document ID: US12006516B2
- Status: Granted
- Kind: B2
- Application number: 18/391,799
- Filed: 2023-12-21
- Published: 2024-04-11
- Granted: 2024-06-11
- Earliest claimed: 2020-07-31
- Assignees: Vertex Pharmaceuticals Incorporated
- Applicants: Vertex Pharmaceuticals Incorporated
- Inventors: George Harb; Chunhi Xie
- Web page: https://patentgrep.com/patent/US12006516B2

## CPC

- C12N 5/0676: {Pancreatic cells}
- C12N 5/0678: {Stem cells; Progenitor cells; Precursor cells}
- A61K 35/39: Pancreas; Islets of Langerhans (Langerhans cells of epidermis A61K35/36)
- C12N 2501/11: Epidermal growth factor [EGF]
- C12N 2501/15: Transforming growth factor beta (TGF-β)
- C12N 2501/155: Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
- C12N 2501/415: Wnt; Frizzeled
- 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

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic endocrine cells. In some aspects, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some aspects, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

## Claims

### Claim 1

A method of administering a composition to a Type 1 Diabetes subject, wherein the composition comprises a cell population, wherein:
(i) the cell population comprises cells that express C-peptide but do not express VMAT1;
(ii) at least 80% of the cells in the cell population express ISL1;
(iii) at most 35% of the cells in the cell population express VMAT1; and
(iv) less than 6% of the cells in the population are NKX6.1-negative, ISL1-negative cells.

### Claim 2 (depends on 1)

The method of claim 1, wherein at most 30% of the cells in the cell population express VMAT1 and at least 20% of the cells in the population express glucagon.

### Claim 3 (depends on 1)

The method of claim 1, wherein at least 15% of the cells in the cell population express glucagon but do not express somatostatin.

### Claim 4 (depends on 1)

The method of claim 1, wherein at least 4% of the cells in the cell population express somatostatin but do not express glucagon.

### Claim 5 (depends on 1)

The method of claim 1, wherein 30-50% of the cells in the population of cells express C-peptide and ISL1 but not VMAT1.

### Claim 6 (depends on 1)

The method of claim 1, wherein 15-40% of the cells in the population of cells express glucagon but not somatostatin.

### Claim 7 (depends on 1)

The method of claim 1, wherein 15-40% of the cells in the population of cells express glucagon but not somatostatin.

### Claim 8 (depends on 1)

The method of claim 1, wherein:
(i) 30-50% of the cells in the population of cells express C-peptide and ISL1 but not VMAT1;
(ii) 15-40% of the cells in the population of cells express glucagon but not somatostatin; and
(iii) wherein at least 4% of the cells in the cell population express somatostatin but do not express glucagon.

### Claim 9 (depends on 8)

The method of claim 8, wherein 80-90% of the cells in the population of cells express ISL1.

### Claim 10 (depends on 8)

The method of claim 8, wherein 85-95% of the cells in the population of cells express ISL1.

### Claim 11 (depends on 1)

The method of claim 1, wherein 5-35% of the cells in the population of cells express VMAT1 but not C-peptide.

### Claim 12 (depends on 11)

The method of claim 11, wherein 80-90% of the cells in the population of cells express ISL1.

### Claim 13 (depends on 11)

The method of claim 11, wherein 85-95% of the cells in the population of cells express ISL1.

### Claim 14 (depends on 1)

The method of claim 1, wherein 5-15% of the cells in the population of cells express VMAT1 but not C-peptide.

### Claim 15 (depends on 14)

The method of claim 14, wherein 30-50% of the cells in the population of cells express C-peptide and ISL1 but not VMAT1.

### Claim 16 (depends on 14)

The method of claim 14, wherein 30-60% of the cells in the population of cells express C-peptide and ISL1 but not VMAT1.

### Claim 17 (depends on 16)

The method of claim 16, wherein the composition is administered to the subject by intraportal transplanting.

### Claim 18 (depends on 17)

The method of claim 17, wherein 15-40% of the cells in the population of cells express glucagon but not somatostatin.

### Claim 19 (depends on 16)

The method of claim 16, wherein the population of cells comprises cells comprising a disruption in the beta-2-microglobulin gene, wherein the disruption reduces or eliminates expression of the beta-2-microglobulin gene.

### Claim 20 (depends on 19)

The method of claim 19, wherein 15-40% of the cells in the population of cells express glucagon but not somatostatin.

### Claim 21 (depends on 16)

The method of claim 16, wherein the method results in the subject having glucose levels of 80-120 milligrams per deciliter (mg/dl) before meals.

### Claim 22 (depends on 21)

The method of claim 21, wherein 15-40% of the cells in the population of cells express glucagon but not somatostatin.

### Claim 23 (depends on 16)

The method of claim 16, wherein 15-40% of the cells in the population of cells express glucagon but not somatostatin.

### Claim 24 (depends on 1)

The method of claim 1, wherein 30-60% of the cells in the population of cells express C-peptide and ISL1 but not VMAT1.

### Claim 25 (depends on 1)

The method of claim 1, wherein the composition is administered to the subject by intraportal transplanting.

### Claim 26 (depends on 1)

The method of claim 1, wherein the composition is administered in a device, wherein the device is configured to produce and release insulin when implanted into a subject.

### Claim 27 (depends on 1)

The method of claim 1, wherein the population of cells comprises cells comprising a disruption in the beta-2-microglobulin gene, wherein the disruption reduces or eliminates expression of the beta-2-microglobulin gene.

### Claim 28 (depends on 1)

The method of claim 1, wherein the method results in the subject having glucose levels of 80-120 milligrams per deciliter (mg/dl) before meals.

### Claim 29 (depends on 1)

The method of claim 1, wherein 80-90% of the cells in the population of cells express ISL1.

### Claim 30 (depends on 1)

The method of claim 1, wherein 85-95% of the cells in the population of cells express ISL1.