application) This application has 2 components. The first focuses on neuroendocrine cells of the prostate, and a transgenic mouse model (CR2-TAg) of metastatic prostate cancer (CaP) arising from this cell lineage. The functions of neuroendocrine cells in the developing and adult prostate remain incompletely defined. Moreover, the appearance of histologic and biochemical features of neuroendocrine differentiation in human CaP is associated with more aggressive disease and androgen independence. CR2-TAg transgenic mice develop prostatic intraepithelial neoplasia (PIN) by 8 weeks of age. This in situ phase is followed by an invasive phase with eventual metastatic spread to lymph nodes, liver, lung, brain and bone. The distinctiveness of the in situ and invasive phases makes this model useful for characterizing the neuroendocrine cell lineage and for identifying the molecular determinants of invasion of a neuroendocrine cancer. We will use these mice to pursue 3 specific aims: (1). Obtain a molecular signature of neuroendocrine cells following initiation of tumorigenesis, at various stages during progression of this cancer, and in the normal prostate using a combination of Affymetrix GeneChips, in situ hybridization, and a new, generally applicable approach for laser capture microdissection (LCM) that allows recovery of intact mRNA from marked cell populations. (2). Produce neuroendocrine cell lines from CR2-TAg mice. (3). Generate Cre-expressing mice for gene knockouts in prostatic neuroendocrine cells. The second component focuses on the urothelium. The undamaged adult mouse urothelium undergoes very slow turnover, making it difficult to identify the molecular regulators of its renewal. The rapid loss and renewal of the (mouse) urothelium following FimH+-dependent attachment of uropathogenic E. coli (UPEC) provides an opportunity to identify the molecular details of urothelial differentiation/survival, and to obtain new understanding about the molecular pathogenesis of urinary tract infection.
Aim 4 will be to use GeneChips to compare gene expression in the bladders of adult female C57Bl/6 mice at various times after exposure to isogenic FimH+ and FimH- strains of UPEC. The cellular origins of observed changes in expression, prior to exfoliation, and during urothelial repair, will be deciphered. The cellular patterns of expression of selected genes will also be characterized in the normal developing and adult urothelium. We have generated transgenic mice that express Cre recombinase in all layers of the urothelium, from the calyces to the bladder, from at least E16.5 through adulthood.
In aim 5, we will continue our efforts to develop an inducible system for performing gene knockouts in all layers of the urothelium. These mice will allow future tests of the contributions of various genes to urothelial function in health and disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
1R01DK059129-01
Application #
6310764
Study Section
Special Emphasis Panel (ZDK1-GRB-5 (O1))
Program Officer
Mullins, Christopher V
Project Start
2000-09-30
Project End
2003-07-31
Budget Start
2000-09-30
Budget End
2001-07-31
Support Year
1
Fiscal Year
2000
Total Cost
$386,125
Indirect Cost
Name
Washington University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Ippolito, Joseph E; Merritt, Matthew E; Backhed, Fredrik et al. (2006) Linkage between cellular communications, energy utilization, and proliferation in metastatic neuroendocrine cancers. Proc Natl Acad Sci U S A 103:12505-10
Ippolito, Joseph E; Xu, Jian; Jain, Sanjay et al. (2005) An integrated functional genomics and metabolomics approach for defining poor prognosis in human neuroendocrine cancers. Proc Natl Acad Sci U S A 102:9901-6
Hu, Yan; Wang, Ting; Stormo, Gary D et al. (2004) RNA interference of achaete-scute homolog 1 in mouse prostate neuroendocrine cells reveals its gene targets and DNA binding sites. Proc Natl Acad Sci U S A 101:5559-64
Hu, Yan; Ippolito, Joseph E; Garabedian, Emily M et al. (2002) Molecular characterization of a metastatic neuroendocrine cell cancer arising in the prostates of transgenic mice. J Biol Chem 277:44462-74
Mysorekar, Indira U; Mulvey, Matthew A; Hultgren, Scott J et al. (2002) Molecular regulation of urothelial renewal and host defenses during infection with uropathogenic Escherichia coli. J Biol Chem 277:7412-9