Our long-term objective is to elucidate basic mechanisms regulating intestinal mucosal functions, with particular emphasis on epithelial cell proliferation and differentiation, and their modulation by growth factors and hormones. The proposed research will utilize a new in vitro model we have developed, centered on human intestinal epithelial cells conditionally-immortalized with a temperature-sensitive SV40 large tumor antigen (tsFHI cell lines). These cells can be maintained without apparent limits in a proliferative state by culture at 32 degrees C, and induced to differentiate at will by a shift to a higher temperature at which the oncogene is inactivated. Using these cells, we will investigate the role and mechanism of action of recently discovered cell cycle regulators, the cyclin-dependent kinase inhibitors (CKIs), and other yet to be identified cellular components, in the intestinal cell differentiation process.
Our specific aims are: 1) To obtain direct evidence for a key role of the CKI p21/Waf-1/Cip1 in the irreversible loss of proliferative potential that marks the onset of tsFHI cell differentiation; 2) To correlate changes in expression and complex formation of other known CKIs with tsFHI cell growth and differentiation; and 3) To identify, clone, and characterize other cellular components whose expression is specifically altered at the time tsFHI cells are induced to differentiate. The methods that will be used in these studies will include immunoprecipitation, Western and Northern blotting, transfection with wild-type or deleted cDNAs, mRNA differential display by PCR, cDNA cloning, sequencing, and analysis of function of cloned genes. The proposed studies should provide information important to further our understanding of a variety of intestinal physiological activities and disease states, in particular nutrition and carcinogenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK048331-02
Application #
2518385
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1996-09-01
Project End
1999-08-31
Budget Start
1997-09-01
Budget End
1998-08-31
Support Year
2
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Cornell University
Department
Physiology
Type
Schools of Arts and Sciences
DUNS #
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Sorensen, Mathew D; Harper, Jonathan D; Hsi, Ryan S et al. (2013) B-mode ultrasound versus color Doppler twinkling artifact in detecting kidney stones. J Endourol 27:149-53
Sorensen, Mathew D; Bailey, Michael R; Shah, Anup R et al. (2012) Quantitative assessment of shockwave lithotripsy accuracy and the effect of respiratory motion. J Endourol 26:1070-4
Shah, Anup; Harper, Jonathan D; Cunitz, Bryan W et al. (2012) Focused ultrasound to expel calculi from the kidney. J Urol 187:739-43
Rimondi, Erika; Secchiero, Paola; Quaroni, Andrea et al. (2006) Involvement of TRAIL/TRAIL-receptors in human intestinal cell differentiation. J Cell Physiol 206:647-54
Weng, Xing-He; Beyenbach, Klaus W; Quaroni, Andrea (2005) Cultured monolayers of the dog jejunum with the structural and functional properties resembling the normal epithelium. Am J Physiol Gastrointest Liver Physiol 288:G705-17
Quaroni, A; Tian, J Q; Seth, P et al. (2000) p27(Kip1) is an inducer of intestinal epithelial cell differentiation. Am J Physiol Cell Physiol 279:C1045-57
Quaroni, A; Tian, J Q; Goke, M et al. (1999) Glucocorticoids have pleiotropic effects on small intestinal crypt cells. Am J Physiol 277:G1027-40
Tian, J Q; Quaroni, A (1999) Involvement of p21(WAF1/Cip1) and p27(Kip1) in intestinal epithelial cell differentiation. Am J Physiol 276:C1245-58
Tian, J Q; Quaroni, A (1999) Dissociation between growth arrest and differentiation in Caco-2 subclone expressing high levels of sucrase. Am J Physiol 276:G1094-104
Quaroni, A; Paul, E C (1999) Cytocentrin is a Ral-binding protein involved in the assembly and function of the mitotic apparatus. J Cell Sci 112 ( Pt 5):707-18

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