Despite the recent flurry of work in the field of gut stem cells, investigators have yet to demonstrate that a single putative stem cell can be isolated, manipulated, maintained and subsequently differentiate into all specified lineages in either an in vitro or in vivo setting. This project's main goal is to demonstrate that a specific putative stem cell can give rise to long-lived clones that have multi-lineage potential. Our central hypothesis is that refinement of methods to isolate and propagate gut stem/progenitors cells will facilitate the in vitro and in vivo development along the secretory lineage as defined by unique markers. Our goals are to (A) refine the techniques and materials that will promote the growth of clusters or single cells in vitro and in vivo;(B) investigate the contribution of intestinal myofibroblasts to the intestinal stem cell niche;(C) modulate the ability of stem/progenitor cells to form fully mature enteroendocrine cells in both in vivo and in vitro models;and (D) identify and characterize other markers from uncommitted progenitors to mature enteroendocrine cells in mice. To address these goals we propose four specific aims.
Specific Aim 1 : To develop methods that overcome the anchorage dependency of gut epithelial cells, and promotes the survival, growth and differentiation of these cells in both in vivo and in vitro settings.
Specific Aim 2 : To assess the role of pericryptal myofibroblast in modulating proliferation of stem cells and the maturation of enteroendocrine cells in a novel clonogenic cell culture system.
Specific Aim 3 : To modulate the maturation of enteroendocrine cells from isolated uncommitted and committed endocrine progenitors in an in vivo and in vitro model.
Specific Aim 4 : Develop and verify further a group of unique lineage-specific markers that define distinct stages in the progression from uncommitted progenitor cells to fully mature enteroendocrine cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01DK085535-03
Application #
8133100
Study Section
Special Emphasis Panel (ZDK1-GRB-8 (O1))
Program Officer
Carrington, Jill L
Project Start
2009-09-30
Project End
2014-08-31
Budget Start
2011-09-01
Budget End
2012-08-31
Support Year
3
Fiscal Year
2011
Total Cost
$250,854
Indirect Cost
Name
University of California Los Angeles
Department
Pediatrics
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Hong, Sung Noh; Dunn, James C Y; Stelzner, Matthias et al. (2017) Concise Review: The Potential Use of Intestinal Stem Cells to Treat Patients with Intestinal Failure. Stem Cells Transl Med 6:666-676
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
Khalil, Hassan A; Lei, Nan Ye; Brinkley, Garrett et al. (2016) A novel culture system for adult porcine intestinal crypts. Cell Tissue Res 365:123-34
Scott, Andrew; Rouch, Joshua D; Jabaji, Ziyad et al. (2016) Long-term renewable human intestinal epithelial stem cells as monolayers: A potential for clinical use. J Pediatr Surg 51:995-1000
Hong, Sung Noh; Dunn, James C Y; Stelzner, Matthias et al. (2016) Concise Review: The Potential Use of Intestinal Stem Cells to Treat Patients With Intestinal Failure. Stem Cells Transl Med :
Rouch, Joshua D; Scott, Andrew; Lei, Nan Ye et al. (2016) Development of Functional Microfold (M) Cells from Intestinal Stem Cells in Primary Human Enteroids. PLoS One 11:e0148216
Khalil, Hassan A; Lei, Nan Ye; Nie, Wenxian et al. (2015) Primary Myofibroblasts Maintain Short-Term Viability following Submucosal Injection in Syngeneic, Immune-Competent Mice Utilizing Murine Colonoscopy. PLoS One 10:e0127258
Wang, Xinwei; Wei, Liang; Cramer, Julie M et al. (2015) Pharmacologically blocking p53-dependent apoptosis protects intestinal stem cells and mice from radiation. Sci Rep 5:8566
Jabaji, Ziyad; Brinkley, Garrett J; Khalil, Hassan A et al. (2014) Type I collagen as an extracellular matrix for the in vitro growth of human small intestinal epithelium. PLoS One 9:e107814
Lei, Nan Ye; Jabaji, Ziyad; Wang, Jiafang et al. (2014) Intestinal subepithelial myofibroblasts support the growth of intestinal epithelial stem cells. PLoS One 9:e84651

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