presentation from dr. melton

Douglas Melton

Dept of Stem Cell and Regenerative Biology

Harvard Stem Cell Institute

Howard Hughes Medical Institute

Stem cells to make β cells and recreate human Type 1 diabetes

JDRF Spring Research Briefing

insulin

glucagon

pancreaticpolypeptide

somatostatin

Pancreatic islet

ghrelin

insulin

glucagon

pancreaticpolypeptide

somatostatin

Pancreatic islet

ghrelin

Best

Banting

1921

Diabetic dog rescued by injection of islet extract

Type I diabetes

1. Loss of beta cells – insulin dependence(relevance for some Type II diabetics)

2. Autoimmune attack of β cells

Analyze the development of the immune attack to induce tolerance

Make more β cells

Human islets for transplantation

Photo: Dr. Gordon Weir, Joslin Diabetes Center

2. Direct reprogramming

1. Directed differentiation of embryonic stem or iPS cells

How to make pancreatic β cells

3. Control β cell replication

2 ways to make more β cells

ES / iPS cell

Definitive endoderm

Foregut

Pancreas

Endocrine

β cell

Directed differentiation to β cells

Sox 17

FoxA2

Pdx1

Ngn3

Growth factors,Co-culture

Chemical screening

O’nightincubation

Starting ES population 5K Chemical

library 4 days incubation

Sox17 staining

DMSO control Hit compound

HTS image-based analysis

High content chemical screens on hES and mES cells

Lee RubinHSCI

2 compounds induce definitive endoderm (IDEs)

IDE compounds efficiently induce definitiveendoderm

> 70%

>50%

mouse

human

2 ways to make more β cells

ES / iPS cell

Definitive endoderm

Foregut

Pancreas

Endocrine

β cell

Directed differentiation to β cells

Compound 1(IDEs, 80%)

Compound 2( Indolactam V, 45%)

Chen et al, Nat Chem Biol, 2009Borowiak et al, Cell Stem Cell, 2009;

Differentiation

cloning / iPS derivation

Direct reprogramming?

islet

acini

duct

vessel

Pancreatic tissues

Expression of 9 trans factors induces extra islet ins+ cells

Reprogrammed cells express genes essential for β cell function

New insulin+ cells closely resemble β cells

Reprogrammed cells produce and secrete insulin

Type I diabetes

1. Loss of beta cells – insulin dependence

2. Autoimmune attack of β cells

Analyze the development of the immune attack to induce tolerance

Make more β cells

Stem Cells in Regenerative Medicine

Key Components for Type I Diabetes

Diabetic patient specific induced Pluripotent Stem (DiPS) cells

4-6 weeks2 days

DiPS

OCT4

KLF4SOX2

Patient Fibroblast DiPS (10x)

Biopsy

DiPS (40x)

2 ways to make more β cells

ES / iPS cell

Definitive endoderm

Pharyngeal foregut

Thymic epithelial progenitor

Directed differentiation to thymic epithelial cells

Thymic epithelial cell

Pax 9

FoxN1

Recapitulating human Type 1 diabetes in a humanized mouse

DiPS

Thymic Epithelial Cells Pancreatic Beta CellsBlood Cells

• Function of cells population• Disease onset and progression• Treatments or cure

Dale GreinerLenny Schultz

NOD, SCID, IL2-rγGeorge Daley

JDRF

HospitalPharma

InsuranceCo.

Academic (Univ)

Patient’s perspective: Who will cure my disease?

Harvard Stem Cell Institute

Dept of Stem Cell and Regenerative Biology

Howard Hughes Medical Institute

Yuval DorDale GreinerLenny ShultzLee RubinStuart SchreiberGeorge DaleyDerrick RossiMarc Vidal

Malgorzata BorowiakShuibing ChenAdriana TajonarRene MaehrSaranya PurushothamanJoe Zhou