Adult tissue stem cells are believed to be responsible for maintaining tissue homeostasis and regeneration following injury in most tissues. The small intestine is one of the most sensitive organs to radiation-induced damage, which is the major complication in abdominal and pelvic radiotherapy with no effective treatment. The intestinal stem cells (ISCs) are located at or near the bottom of crypts in a unique yet poorly defined microenvironment ("niche") composed of several cell types derived from the bone marrow (BM). However, their identity remained elusive until recently. Genetic evidence demonstrated that the crypt-based columnar cells (CBCs) interspersed among Paneth cells marked by Lgr5 and some +4 cells immediately above Paneth cells marked by Bmi-1, represent perhaps distinct subsets of ISCs. Therefore, how their functions are regulated by intrinsic and extrinsic (niche-related) programs to participate in intestinal regeneration following injury is poorly understood. Work from us and others indicate that the BH3-only protein PUMA and cyclin-dependent kinase inhibitor p21 regulate the survival and regeneration of intestinal and hematopoietic systems, and the bone marrow modulates intestinal radiosensitivity. We hypothesize that both apoptosis and cell cycle arrest critically regulate the survival and regeneration of intestinal stem cells following radiation through intrinsic and extrinsic mechanisms. In this proposal, we will combine a novel ISC lineage marking and tracing mouse model with various knockout and transplantation models to 1) define the role of ISCs in crypt regeneration following radiation and develop assays for their isolation and characterization;2) demonstrate a potential coordination of PUMA and p21 in modulating the survival and regeneration of ISCs;and 3) define the potential bone marrow contributions to intestinal regeneration. We believe that our studies can lead to a better understanding of ISC biology, and new models and strategies for isolation, characterization and manipulation of these critical cells for therapeutic purposes.

Public Health Relevance

The proposed studies aim to determine the underlying molecular mechanisms governing the injury, survival and regeneration of intestinal stem cells following radiation, and to develop assays for their isolation and characterization. Such studies can help devise strategies to combat intestinal damage caused by radiotherapy or accidental radiation exposure.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project--Cooperative Agreements (U01)
Project #
3U01DK085570-05S1
Application #
8700642
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
2013-09-01
Budget End
2014-08-31
Support Year
5
Fiscal Year
2013
Total Cost
$79,300
Indirect Cost
$27,300
Name
University of Pittsburgh
Department
Pathology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Hartman, Kira G; Bortner Jr, James D; Falk, Gary W et al. (2014) Modeling human gastrointestinal inflammatory diseases using microphysiological culture systems. Exp Biol Med (Maywood) 239:1108-23
Zhang, Lin; Yu, Jian (2013) Role of apoptosis in colon cancer biology, therapy, and prevention. Curr Colorectal Cancer Rep 9:
Qiu, W; Liu, H; Sebastini, A et al. (2013) An apoptosis-independent role of SMAC in tumor suppression. Oncogene 32:2380-9
Spender, Lindsay C; Carter, Matthew J; O'Brien, Darren I et al. (2013) Transforming growth factor-? directly induces p53-up-regulated modulator of apoptosis (PUMA) during the rapid induction of apoptosis in myc-driven B-cell lymphomas. J Biol Chem 288:5198-209
Qiu, W; Wang, X; Buchanan, M et al. (2013) ADAR1 is essential for intestinal homeostasis and stem cell maintenance. Cell Death Dis 4:e599
Magness, Scott T; Puthoff, Brent J; Crissey, Mary Ann et al. (2013) A multicenter study to standardize reporting and analyses of fluorescence-activated cell-sorted murine intestinal epithelial cells. Am J Physiol Gastrointest Liver Physiol 305:G542-51
Stelzner, Matthias; Helmrath, Michael; Dunn, James C Y et al. (2012) A nomenclature for intestinal in vitro cultures. Am J Physiol Gastrointest Liver Physiol 302:G1359-63
Li, Hua; Wang, Peng; Yu, Jian et al. (2011) Cleaving Beclin 1 to suppress autophagy in chemotherapy-induced apoptosis. Autophagy 7:1239-41
Dirisina, Ramanarao; Katzman, Rebecca B; Goretsky, Tatiana et al. (2011) p53 and PUMA independently regulate apoptosis of intestinal epithelial cells in patients and mice with colitis. Gastroenterology 141:1036-45
Wang, Yu Fu; Xu, Xiang; Fan, Xia et al. (2011) A cell-penetrating peptide suppresses inflammation by inhibiting NF-ýýB signaling. Mol Ther 19:1849-57

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