): The objectives of this proposal are to a) provide a structured, training program in molecular genetics for the applicant to develop into an independent researcher, and b) study the genetic events associated with overcoming senescence (i.e. immortalization) in human prostate epithelial cells. Numerous documentational studies have detailed patterns of genomic loss and gain in prostate cancers. However, the functional significance of many of these genetic events or combinations of genetic events remains unclear. Overcoming senescence can be considered a critical event in tumorigenesis that has a clear genetic basis. In the proposed model, human prostate epithelial cultures (HPECs) expressing viral oncoproteins lack normal pRB and/or p53 regulatory functions (genes commonly altered in clinical prostate cancers) and have an extended lifespan. These in vitro events place these cells at high risk for additional 'spontaneous' genetic and epigenetic alterations associated with immortalization. The applicants will test the hypothesis that combinations of these genetic events, complementing p53 and/or pRb loss, are important in overcoming cellular senescence in in vitro prostate cancer. In addition, they propose that these pathways are found in clinical prostate cancer specimens.
Their SPECIFIC AIMS i nclude: I) to establish and characterize an in vitro model system with immortal HPECs that have functionally lost p53 and/or Rb by selective HPV16 E6 and/or E7 retroviral infection, ii) to identify additional genetic and epigenetic events, chiefly combinations of events, associated with overcoming senescence for p53- and Rb-linked pathways, iii) to reexpress these genomic regions lost or gained at immortalization in immortal and normal HPECs, and iv) to correlate these in vitro events with clinical prostate cancer samples that have lost either p53 and/or Rb function. The proposal will provide new i n sight into molecular genetic and epigenetic events associated with overcoming senescence, via p53 and/or Rb loss of function, in prostate epithelial cells in vitro. In addition to addressing this critical mechanistic role in human prostate neoplasia, it will provide the applicant with a structured training program in molecular genetics in the laboratory of Dr. Catherine Reznikoff.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08CA076184-04
Application #
6350257
Study Section
Subcommittee G - Education (NCI)
Program Officer
Myrick, Dorkina C
Project Start
1998-02-01
Project End
2003-01-31
Budget Start
2001-02-01
Budget End
2002-01-31
Support Year
4
Fiscal Year
2001
Total Cost
$92,070
Indirect Cost
Name
University of Wisconsin Madison
Department
Surgery
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Schwarze, Steven R; Fu, Vivian X; Desotelle, Joshua A et al. (2005) The identification of senescence-specific genes during the induction of senescence in prostate cancer cells. Neoplasia 7:816-23
Fu, Vivian X; Schwarze, Steven R; Kenowski, Michelle L et al. (2004) A loss of insulin-like growth factor-2 imprinting is modulated by CCCTC-binding factor down-regulation at senescence in human epithelial cells. J Biol Chem 279:52218-26
Schwarze, Steven R; Fu, Vivian X; Jarrard, David F (2003) Cdc37 enhances proliferation and is necessary for normal human prostate epithelial cell survival. Cancer Res 63:4614-9
Jarrard, David F; Modder, Joshua; Fadden, Paul et al. (2002) Alterations in the p16/pRb cell cycle checkpoint occur commonly in primary and metastatic human prostate cancer. Cancer Lett 185:191-9
Schwarze, Steven R; DePrimo, Samuel E; Grabert, Lisa M et al. (2002) Novel pathways associated with bypassing cellular senescence in human prostate epithelial cells. J Biol Chem 277:14877-83
Schwarze, S R; Shi, Y; Fu, V X et al. (2001) Role of cyclin-dependent kinase inhibitors in the growth arrest at senescence in human prostate epithelial and uroepithelial cells. Oncogene 20:8184-92
Kinoshita, H; Shi, Y; Sandefur, C et al. (2000) Methylation of the androgen receptor minimal promoter silences transcription in human prostate cancer. Cancer Res 60:3623-30
Kinoshita, H; Shi, Y; Sandefur, C et al. (2000) Screening hypermethylated regions by methylation-sensitive single-strand conformational polymorphism. Anal Biochem 278:165-9
Jarrard, D F; Sarkar, S; Shi, Y et al. (1999) p16/pRb pathway alterations are required for bypassing senescence in human prostate epithelial cells. Cancer Res 59:2957-64
Jarrard, D F; Kinoshita, H; Shi, Y et al. (1998) Methylation of the androgen receptor promoter CpG island is associated with loss of androgen receptor expression in prostate cancer cells. Cancer Res 58:5310-4