Maintenance of an intact intestinal epithelium is critical for intestinal growth, development, wound healing and prevention of disease. Restitution, the initial phase of epithelial wound repair, is regulated by a number of peptides, including epidermal growth factor (EGF). Altered mucosal reepithelialization is a significant component of a number of diseases primarily affecting the gastrointestinal tract, such as ulcerative colitis and peptic ulcer disease. Surprisingly, little information is known about the initial signaling events in the process of basal or stimulated intestinal cell migration. Preliminary data in intestinal epithelial cells from our laboratory and reports on fibroblast migration have permitted the development of the proposed hypothesis that: Epidermal growth factor receptor regulates intestinal epithelial cell migration via integrated signaling pathways requiring tyrosine kinase and the enhanced activities of specific target molecules during the earliest phases of cellular movement. The objective of this proposal is to understand the cellular and molecular mechanisms of growth factor regulated intestinal cell migration through a series of questions comprising the following Specific Aims: 1) What are the principal mechanisms of EGF receptor regulation of intestinal epithelial cell migration? 2) What are the roles of phosphatidylinositol phospholipase C (PLC) gamma 1, protein kinase (PK) C epsilon, and other signal transduction molecules in EGF receptor-regulated cellular migration? and 3) How does the EGF receptor mediate cytoskeletal rearrangement during intestinal cell migration? To address the mechanisms of EGF receptor regulation of migration in aim 1, we will express various EGF receptor constructs in a mouse colon cell line derived from the EGF receptor null mouse.
In aims 2 and 3, we will express constitutively active and dominant negative forms of PLC gamma 1, PKC epsilon and the Rho small GTPase proteins to establish a molecular ordering of signal transduction from the EGF receptor to cytoskeletal rearrangement regulating motility. We will use expression based cloning and coprecipitation assays to identify interacting proteins in the pathway leading to cellular migration. These studies should enhance our understanding of the initial signaling events required for growth factor-regulated intestinal epithelial restitution and provide a mechanistic basis for potential development of novel therapeutic targets.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK054993-04
Application #
6771686
Study Section
General Medicine A Subcommittee 2 (GMA)
Program Officer
May, Michael K
Project Start
2001-07-01
Project End
2006-06-30
Budget Start
2004-07-01
Budget End
2005-06-30
Support Year
4
Fiscal Year
2004
Total Cost
$221,970
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Pediatrics
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Dubé, Philip E; Liu, Cambrian Y; Girish, Nandini et al. (2018) Pharmacological activation of epidermal growth factor receptor signaling inhibits colitis-associated cancer in mice. Sci Rep 8:9119
Shen, Xi; Liu, Liping; Peek, Richard M et al. (2018) Supplementation of p40, a Lactobacillus rhamnosus GG-derived protein, in early life promotes epidermal growth factor receptor-dependent intestinal development and long-term health outcomes. Mucosal Immunol 11:1316-1328
Miguel, Jennifer C; Maxwell, Adrienne A; Hsieh, Jonathan J et al. (2017) Epidermal growth factor suppresses intestinal epithelial cell shedding through a MAPK-dependent pathway. J Cell Sci 130:90-96
Yan, F; Liu, L; Cao, H et al. (2017) Neonatal colonization of mice with LGG promotes intestinal development and decreases susceptibility to colitis in adulthood. Mucosal Immunol 10:117-127
Wang, Y; Liu, L; Moore, D J et al. (2017) An LGG-derived protein promotes IgA production through upregulation of APRIL expression in intestinal epithelial cells. Mucosal Immunol 10:373-384
Zhao, Gang; Liu, Liping; Peek Jr, Richard M et al. (2016) Activation of Epidermal Growth Factor Receptor in Macrophages Mediates Feedback Inhibition of M2 Polarization and Gastrointestinal Tumor Cell Growth. J Biol Chem 291:20462-72
Dubé, Philip E; Punit, Shivesh; Polk, D Brent (2015) Redeeming an old foe: protective as well as pathophysiological roles for tumor necrosis factor in inflammatory bowel disease. Am J Physiol Gastrointest Liver Physiol 308:G161-70
Punit, Shivesh; Dubé, Philip E; Liu, Cambrian Y et al. (2015) Tumor Necrosis Factor Receptor 2 Restricts the Pathogenicity of CD8(+) T Cells in Mice With Colitis. Gastroenterology 149:993-1005.e2
Lu, Ning; Wang, Lihong; Cao, Hailong et al. (2014) Activation of the epidermal growth factor receptor in macrophages regulates cytokine production and experimental colitis. J Immunol 192:1013-23
Sierra, Johanna C; Hobbs, Stuart; Chaturvedi, Rupesh et al. (2013) Induction of COX-2 expression by Helicobacter pylori is mediated by activation of epidermal growth factor receptor in gastric epithelial cells. Am J Physiol Gastrointest Liver Physiol 305:G196-203

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