The Stat family of transcription factors is necessary for the growth and differentiation of mammary tissues. In mammary tissues, a significant means of Stat activation is the binding of the neuroendocrine hormone prolactin (PRL) to its receptor, the PRLr. PRL/PRLr binding results in the activation of receptor-associated signaling networks, such as the Jak/Stat pathway and the endocytosis and nuclear retrotransport of a complex between PRL and the prolyl isomerase cyclophilin B (CypB). The intranuclear PRL/CypB complex regulates Stat5-mediated gene expression and DNA binding. Using multiple approaches including yeast two-hybrid and transcription factor array analysis, we have demonstrated that the PRL/CypB complex regulates the interaction of Stat5 with multiple intranuclear proteins including the peptide inhibitor of activated Stat, PIAS3, the small ubiquitin-related modifier, SUMO2, and the transcription factor and proto-oncogene, c-Myb. It is the central hypothesis of this proposal that the temporal and spatial interaction of Stat5 with these proteins, as modulated by the PRL/CypB complex, regulates Stat5 function. This hypothesis will be tested by three specific aims using breast cancer and PRL-responsive cell lines both in vitro and in vivo. First, the structural basis for Stat5/PIAS3 interaction and the functional significance of this interaction will be mapped using complementary mutagenesis/overexpression and knockdown strategies. Second, the functional consequences of Stat5 sumolyation will be evaluated by cellular and biochemical approaches. Third, the association between Stat5 and co-activators/repressors, such as c-Myb will be assessed by targeted mutagenesis, small-interfering RNA (SiRNA), and chromatin immunoprecipitation (ChlP)-based methodologies. Given that our preliminary evidence indicates that the manipulation of these interactions results in the alteration of Stat5-mediated gene expression and the survival of breast cancer cells, our proposed studies are directly relevant to a better understanding of the pathophysiology of: human breast cancer, and as such, may provide the basis for novel therapeutic strategies aimed at modifying Stat function in this disease. ? ?

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA102682-05
Application #
7224178
Study Section
Reproductive Endocrinology Study Section (REN)
Program Officer
Knowlton, John R
Project Start
2003-07-01
Project End
2009-04-30
Budget Start
2007-05-01
Budget End
2009-04-30
Support Year
5
Fiscal Year
2007
Total Cost
$246,034
Indirect Cost
Name
Northwestern University at Chicago
Department
Pathology
Type
Schools of Medicine
DUNS #
005436803
City
Chicago
State
IL
Country
United States
Zip Code
60611
Medler, Terry R; Craig, Justin M; Fiorillo, Alyson A et al. (2016) HDAC6 Deacetylates HMGN2 to Regulate Stat5a Activity and Breast Cancer Growth. Mol Cancer Res 14:994-1008
Zheng, Jiamao; Fang, Feng; Zeng, Xianke et al. (2011) Negative cross talk between NFAT1 and Stat5 signaling in breast cancer. Mol Endocrinol 25:2054-64
Fang, Feng; Zheng, Jiamao; Galbaugh, Traci L et al. (2010) Cyclophilin B as a co-regulator of prolactin-induced gene expression and function in breast cancer cells. J Mol Endocrinol 44:319-29
Clevenger, Charles V; Gadd, Samantha L; Zheng, Jiamao (2009) New mechanisms for PRLr action in breast cancer. Trends Endocrinol Metab 20:223-9
Fang, Feng; Flegler, Ayanna J; Du, Pan et al. (2009) Expression of cyclophilin B is associated with malignant progression and regulation of genes implicated in the pathogenesis of breast cancer. Am J Pathol 174:297-308
Fang, Feng; Rycyzyn, Michael A; Clevenger, Charles V (2009) Role of c-Myb during prolactin-induced signal transducer and activator of transcription 5a signaling in breast cancer cells. Endocrinology 150:1597-606
McHale, Kevin; Tomaszewski, John E; Puthiyaveettil, Ragunath et al. (2008) Altered expression of prolactin receptor-associated signaling proteins in human breast carcinoma. Mod Pathol 21:565-71
Zheng, Jiamao; Koblinski, Jennifer E; Dutson, Laura V et al. (2008) Prolyl isomerase cyclophilin A regulation of Janus-activated kinase 2 and the progression of human breast cancer. Cancer Res 68:7769-78
Clevenger, Charles V; Zheng, Jiamao; Jablonski, Elizabeth M et al. (2008) From bench to bedside: future potential for the translation of prolactin inhibitors as breast cancer therapeutics. J Mammary Gland Biol Neoplasia 13:147-56
Fang, Feng; Antico, Giovanni; Zheng, Jiamao et al. (2008) Quantification of PRL/Stat5 signaling with a novel pGL4-CISH reporter. BMC Biotechnol 8:11

Showing the most recent 10 out of 12 publications