human development - columbia universitypancreasdev.ppt author: sussell created date: 3/11/2009...
TRANSCRIPT
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Pancreas and Liver Development
Human Development
Lori Sussel, PhD Department of Genetics and Development
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Location of the pancreas and liver
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Pancreas
• gland responsible for energy homeostasis
• development has been major focus of research over the past 15 years
Islet (endocrine)
exocrine
duct
insulin, amylase
H&E
exocrine
islet
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Pancreatic Cell Types
Edlund, 2002 ε cells
Exocrine (85%)
Endocrine (10%) (Islets)
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Cancer: disease of the exocrine pancreatic ducts
• 95 percent of pancreatic cancers start in the exocrine ductal cells
• diagnosed in ~ 30,000 people in the US each year • 4th leading cause of cancer-related deaths • often no symptoms early on; difficult to diagnose in its
beginning stages; most pancreatic cancers have spread beyond gland by diagnosis
• high mortality rate • pancreatic tumors have the poorest responses to
treatment among all the major cancers
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Diabetes: Disease of the endocrine pancreas
180 Million Worldwide~2000 360 Million Worldwide~2030
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Organogenesis of the Pancreas
• arises from foregut endoderm • initially forms as two separate and distinct rudiments
which fuse to form a single organ containing all cell types
• mammals, birds, reptiles, amphibians and zebrafish have a pancreas with similar histology and mode of development
• organogenesis depends on complex interactions between epithelium and mesenchyme
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Murtaugh, 2007
Overview of pancreas development
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Pdx1
Exocrine tissue = acinar cells
Endocrine tissue = islet cells
Definitive pancreas marker
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Pancreas development
Offield et al., 1996
9.5 dpc 11.5 dpc 16.5 dpc
v
d d
v
du
Pdx1:LacZ
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Pancreas development
Wells and Melton, 1999
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Stages of pancreas development
9.5 dpc 14.5-16.5 10.5 - 14.5 16.5 - 18.5
Panc. bud evagination
exocrine & endocrine
differentiation
islet formation
endocrine differentiation
26 dpc 30-60 dpc 25-29 wpc 12-20 wpc
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Pancreas Looping
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Published by AAAS K. S. Zaret et al., Science 322, 1490 -1494 (2008)
Early patterning of the endoderm
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Published by AAAS K. S. Zaret et al., Science 322, 1490 -1494 (2008)
Early patterning of the endoderm
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Notochord
Pancreatic endoderm
E8.5-9.0
At e8.0, dorsal pancreatic endoderm is induced by the notochord
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Experiment: Remove the notochord and see what
happens to the pancreas
Kim, Hebrok and Melton, 1997-1999
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Shh expression
Notochord
Kim, Hebrok and Melton, 1997-1999
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Notochord is necessary to specify pancreas
Kim, Hebrok and Melton, 1997-1999
Pancreas markers
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Notochord is sufficient to specify dorsal pancreas
Kim, Hebrok and Melton, 1997-1999
Negative controls
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Notochord activin FGF2
ActR FGFR Dorsal
pancreatic endoderm
Shh
Pdx1
Signaling pathway directed by notochord
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What patterns the ventral pancreas?
\ DP
VP
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Published by AAAS K. S. Zaret et al., Science 322, 1490 -1494 (2008)
Early patterning of the endoderm
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Specification of ventral pancreas linked to liver specification
Zaret, 2002
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Ventral pancreas induction
• Does not receive signals from notochord or dorsal aorta
• Develops next to cardiac mesoderm
• FGF and BMP signals from cardiac mesoderm required for liver induction and restriction of ventral pancreas domain (Zaret)
• Shh is activated (opposite from dorsal)
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Kim and MacDonald, 2002
M M
M M
Pancreatic Mesenchyme
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Pancreatic Mesenchyme • Mesoderm accumulates around pancreatic
epithelial buds • Mesenchyme is necessary for
cytodifferentiation and morphogenesis (Golosow and Grobstein, 1962)
• Signaling is permissive – FGF10 – Notch – TGFβ family – Wnts
• Activation of pancreas transcriptional program
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Mesenchymal signals are necessary for pancreatic growth and differentiation
G. Gittes
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Mesenchymal signals: Time and space dependent
• Early experiments suggested endocrine was default lineage – Early mesenchyme favors endocrine
development – Late mesenchyme favors exocrine
• Contact dependent signaling – proexocrine factor(s): cell-contact mediated – proendocrine factor(s): diffusible
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Signaling pathways: what molecules are involved?
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Murtaugh and Melton, 2003 Fgf Activin
Fgf Notch
Notch
????? Shh
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Bhushan, A. et al. Development 2001;128:5109-5117
The size of the pancreatic epithelium in Fgf10-/- embryos is greatly reduced
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Transcriptional control of pancreatic differentiation
Signaling events culminate in activation of transcriptional program
Transcription factor studies highlight several new and traditional mouse manipulation
techniques
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Pancreatic cell type specification
Pancreas Progenitor
Pdx1 Ptf1a
ductal exocrine
Endocrine Progenitor
Pdx1 Ngn3
Ptf1a ?
Notch
α
β
δ
PP
glucagon
insulin
somatostatin
pancreatic polypeptide
ε ghrelin
Signals ?????
Pdx1 NeuroD1 Nkx2.2 Pax4 Nkx6.1 Hlxb9 Pax6
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Pdx1 Pancreatic Duodenal Homeobox 1
• Also known as IPF1, STF1, IDX1 • Expression identifies region of pancreas
specification prior to visible morphological changes
• Earliest and one of the most specific genes expressed in pancreatic primordia
• Functions at several time points during pancreas development
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Pdx1 expression
Offield et al., 1996
9.5 dpc 11.5 dpc 16.5 dpc
v
d d
v
du
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Pdx1 expression
• Throughout early pancreatic epithelium • Pancreas progenitors • β and δ cells (high levels) • Exocrine cells (low levels)
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Pdx1 null causes pancreatic agenesis
Offield et al., 1996
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Pdx1 mutations in humans
• Loss of function mutations cause apancreatic phenotype and perinatal lethality (failure to thrive infants)
• Reduced function mutations: MODY4 – MODY = Maturity onset diabetes of the
young
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Pdx1
Exocrine
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Ngn3 is expressed in endocrine progenitors
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Copyright ©2000 by the National Academy of Sciences
Gradwohl, Gérard et al. (2000) Proc. Natl. Acad. Sci. USA 97, 1607-1611
Ngn3 null: all islet lineages are lost
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Ngn3 summary
• Ngn3 is expressed in the endocrine progenitor cells
• Ngn3 cells can give rise to all the islet cell populations
• The islet progenitor cells are differentially competent over time to give rise to the different islet cell types
• Reactivated during pancreas regeneration
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Murtaugh, 2007
Summary of pancreas development
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Liver
• Largest internal organ in the body
• Two major lobes
• Hepatocytes (60-80% of liver cells) carry out main functions of the liver
• Many functions including fat breakdown, filtration, vitamin storage, glucose regulation, cholesterol production
• Genetic liver diseases, hepatitis, cirrhosis
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Published by AAAS K. S. Zaret et al., Science 322, 1490 -1494 (2008)
Liver derived next to the v. pancreas
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Liver organogenesis • Derived from endoderm layer as a single rudiment • Requires a series of inductive signals from at least 3
different mesodermal cell types • Begins forming at e8.5 when hepatic epithelium
thickens, delaminates and invades surrounding mesenchyme to form the liver bud
• Endothelial cells critical for liver development and differentiation
• Continued epithelial-mesenchymal interactions stimulate cell proliferation and morphogenesis as the organ grows
• High regenerative capacity (replication of existing cell types)
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Sequential stages of liver development
Endoderm patterning and
hepatic competence (gastrulation)
Liver bud proliferation
morphogenesis
Liver induction and specification (4-8 somite)
Hepatic differentiation
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Establishment of competence and specification
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Cell type differentiation
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Bud formation
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Published by AAAS K. S. Zaret et al., Science 322, 1490 -1494 (2008)
Progenitor cells in liver and pancreas
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Stem cells --> Islet cells
pancreas
brain liver
or
ES cells
?
?
?
iPS cells
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