presentation from dr. melton
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![Page 1: Presentation from Dr. Melton](https://reader033.vdocuments.net/reader033/viewer/2022051314/54c62c974a7959390f8b457d/html5/thumbnails/1.jpg)
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
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insulin
glucagon
pancreaticpolypeptide
somatostatin
Pancreatic islet
ghrelin
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insulin
glucagon
pancreaticpolypeptide
somatostatin
Pancreatic islet
ghrelin
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Best
Banting
1921
Diabetic dog rescued by injection of islet extract
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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
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Human islets for transplantation
Photo: Dr. Gordon Weir, Joslin Diabetes Center
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2. Direct reprogramming
1. Directed differentiation of embryonic stem or iPS cells
How to make pancreatic β cells
3. Control β cell replication
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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
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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
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2 compounds induce definitive endoderm (IDEs)
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IDE compounds efficiently induce definitiveendoderm
> 70%
>50%
mouse
human
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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;
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Differentiation
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cloning / iPS derivation
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Direct reprogramming?
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islet
acini
duct
vessel
Pancreatic tissues
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Expression of 9 trans factors induces extra islet ins+ cells
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Reprogrammed cells express genes essential for β cell function
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New insulin+ cells closely resemble β cells
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Reprogrammed cells produce and secrete insulin
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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
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Stem Cells in Regenerative Medicine
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Key Components for Type I Diabetes
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Diabetic patient specific induced Pluripotent Stem (DiPS) cells
4-6 weeks2 days
DiPS
OCT4
KLF4SOX2
Patient Fibroblast DiPS (10x)
Biopsy
DiPS (40x)
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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
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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
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JDRF
HospitalPharma
InsuranceCo.
Academic (Univ)
Patient’s perspective: Who will cure my disease?
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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