Deriving Complex Endodermal Organ Tissues from Human Pluripotent Stem Cells As Models for Development and Disease Research

Kyle
McCracken*, Jorge Munera*, Patrick McGrath*, Michael
Workman*, Katie Sinagoga*, Christopher Mayhew*, Noah Shroyer⁺, Carey Watson^, Michael Helmrath^ and James Wells^#*.

Divisions of Developmental
Biology*, Gastroenterology+,  Endocrinology#, Pediatric Surgery^

Cincinnati ChildrenÕs
Hospital Medical Center, Cincinnati OH 45229

Successful
efforts to direct the differentiation of
human
embryonic and induced pluripotent stem
cells
(PSCs) into specific organ cell
types in vitro have largely been guided by
studies
in embryonic development. We have used signaling pathways that control early endoderm
organ morphogenesis in vivo to
generate complex, three-dimensional organ tissues with improved functionality from
human PSCs in vitro. We identified that the combined
activity of FGF and Wnt signaling pathways was
sufficient to promote PSC-derived endoderm to undergo gut tube morphogenesis in vitro. The resulting
three-dimensional gut tube tissues can be further directed into specific organ
tissue types using embryonic patterning pathways. For example we have been able
to use a temporal series of growth factor manipulations that mimic embryonic
intestinal development to generate three-dimensional human intestinal organoids (HIOs). HIOs consisted of a polarized, columnar
epithelium with villus-like structures and crypt-like proliferative zones with
all of the major cell types of the adult gut. Cell types within HIOs can be
further using genetic gain- and loss-of-function approaches, or by adding
progenitors of other lineages during the process of in vitro development. We have used similar
three-dimensional approaches to generate human foregut organoids
capable of giving rise to lung, pancreas, and stomach tissues. We have directed
differentiation of human foregut tissue specifically into stomach/gastric organoids, which we are using to model the pathogenesis
induced by infection with Helicobacter
pylori. We are also using genetically modified PSCs, or iPSCs
derived from patients, to establish models of disease including cystic fibrosis
and malabsorption syndromes. 

Grant
funding: NIH, 1U18NS080815,
R01DK080823 and S1 (J.W.), 1R01DK092456 (J.W. and
N.S) and a CTSA (U54 RR025216 01).