The tremendous regenerative capacity of the intestine, with epithelial turnover every 5-7 days, is mediated by the carefully orchestrated activity of intestinal stem cells (ISC). The recent identification of molecular markers for ISCs, combined with in vivo lineage tracing and in vitro primary organoid culture methodologies has revolutionized ISC biology. The resultant wealth of findings have showcased ISCs as a robust general paradigm for stem cell behavior and have elicited a next generation of studies exploring the ISC niche, interrelationships and interconversion of actively cycling versus quiescent ISC populations, and translational applications of ISC transplantation. Accordingly, the current application is a continuation of our prior 5 year project for the NIDDK Intestinal Stem Cell Consortium (ISCC), and is responsive to RFA-DK-13-012, Intestinal Stem Cell Consortium Research Projects (U01), seeking to """"""""define conditions controlling proliferation and differentiation of intestinal stem cells"""""""", """"""""determine the developmental lineage of characterized populations of stem cells in the small intestine"""""""" and """"""""methods for expanding and grafting stem cells back into the intestine"""""""". Here, we build upon our preliminary data from the prior cycle with novel reagents and methodologies.
Aim 1 explores R-spondins as regulators of Lgr5+ ISC symmetric cell division combining adenovirus-mediated gain- and loss-of-function approaches with in vivo lineage tracing, and pursuing mechanistic and niche localization studies.
In Aim 2, interrelationships between diverse cycling and quiescent ISC populations will be investigated by RNA-Seq, and novel lineage relationships of Bmi1+ ISC will be explored.
Aims 3 and 4 explore translational aspects of ISC biology, with Aim 3 utilizing organoid culture to evaluating the effects of R- spondins on colon cancer stem cells, and Aim 4 attempting the phenotypic correction of a Mendelian intestinal epithelial defect by ISC transplantation. Overall, these studies describe an integrated approach to ISC biology and translation that is highly responsive to RFA-DK-13-012, Intestinal Stem Cell Consortium Research Projects (U01), with implications for stem cell biology, and intestinal disease pathogenesis and therapy.

Public Health Relevance

This project uses genetic and cell culture methods to investigate how intestinal stem cells divide under normal conditions, and are activated under injury conditions. A further goal is to utilize intestinal stem cell biology for treatment of colon cancer and for intestinal stem cell transplantation to correct genetic disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project--Cooperative Agreements (U01)
Project #
2U01DK085527-06
Application #
8774782
Study Section
Special Emphasis Panel (ZDK1)
Program Officer
Carrington, Jill L
Project Start
2009-09-22
Project End
2019-08-31
Budget Start
2014-09-01
Budget End
2015-08-31
Support Year
6
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Stanford University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Stanford
State
CA
Country
United States
Zip Code
94304
Vallon, Mario; Yuki, Kanako; Nguyen, Thi D et al. (2018) A RECK-WNT7 Receptor-Ligand Interaction Enables Isoform-Specific Regulation of Wnt Bioavailability. Cell Rep 25:339-349.e9
Yan, Kelley S; Gevaert, Olivier; Zheng, Grace X Y et al. (2017) Intestinal Enteroendocrine Lineage Cells Possess Homeostatic and Injury-Inducible Stem Cell Activity. Cell Stem Cell 21:78-90.e6
Janda, Claudia Y; Dang, Luke T; You, Changjiang et al. (2017) Surrogate Wnt agonists that phenocopy canonical Wnt and ?-catenin signalling. Nature 545:234-237
Yan, Kelley S; Janda, Claudia Y; Chang, Junlei et al. (2017) Non-equivalence of Wnt and R-spondin ligands during Lgr5+ intestinal stem-cell self-renewal. Nature 545:238-242
Chang, Junlei; Mancuso, Michael R; Maier, Carolina et al. (2017) Gpr124 is essential for blood-brain barrier integrity in central nervous system disease. Nat Med 23:450-460
Mah, Amanda T; Kuo, Calvin J (2016) Home Sweet Home: a Foxl1+Mesenchymal Niche for Intestinal Stem Cells. Cell Mol Gastroenterol Hepatol 2:116-117
Li, Xingnan; Ootani, Akifumi; Kuo, Calvin (2016) An Air-Liquid Interface Culture System for 3D Organoid Culture of Diverse Primary Gastrointestinal Tissues. Methods Mol Biol 1422:33-40
Mah, Amanda T; Yan, Kelley S; Kuo, Calvin J (2016) Wnt pathway regulation of intestinal stem cells. J Physiol 594:4837-47
DiMarco, Rebecca L; Dewi, Ruby E; Bernal, Gabriela et al. (2015) Protein-engineered scaffolds for in vitro 3D culture of primary adult intestinal organoids. Biomater Sci 3:1376-85
Huang, Julie Y; Sweeney, Emily Goers; Sigal, Michael et al. (2015) Chemodetection and Destruction of Host Urea Allows Helicobacter pylori to Locate the Epithelium. Cell Host Microbe 18:147-56

Showing the most recent 10 out of 33 publications