Tumor metastasis is responsible for most treatment failures in breast cancer patients. In cancers of epithelial origin, loss of epithelial cell polarity and transformation into a migratory phenotype are key steps in invasion of cancer cells. Intravasation of cancer cells appears to be dependent upon epidermal growth factor (EGF)-stimulated chemotaxis. Chemotaxis and invasion depends upon dynamic adhesion to extracellular matrix and actin cytoskeleton remodeling. Phosphoinositide signaling plays a key role in these processes, but the underlying mechanisms are poorly defined. Type Igamma phosphatidylinositol-4-phosphate 5-kinase (PIPKIgamma) synthesizes phosphatidylinositol-4,5- bisphosphate (PIP2) in a temporal and spatial fashion within cells. Hypothesis: PIPKIgamma isoforms, via their expression, targeting, and regulation by EGF-stimulation, generate the messenger PIP2, which modulate the spatial and temporal assembly of focal adhesions and vesicular trafficking that regulates chemotaxis. The role of PIPKIgamma in regulating migration is key in the metastasis of tumors.
Specific Aims : (1) The mechanistic role of PIPKIgamma in growth factor-stimulated directional migration and invasion will be investigated. EGF signaling mechanisms leading to phosphorylation of PIPKIgamma will be characterized as will the role of PIP2 production in regulating tyrosine phosphorylation of PIPKIgamma and other proteins at focal adhesions. The underlying mechanism for PIPKgamma participation in EGF- stimulated chemotaxis and invasion will be explored, with an emphasis on the dynamic assembly of focal adhesions (FA) and vesicular trafficking. GFP/RFP-focal adhesion proteins will be used to define the role of PIPKIgamma in focal adhesions dynamics in EGF-stimulated cells. (2) Two new alternatively spliced PIPKIgamma isoforms that express unique C-terminal extensions will be characterized. PIPKIgamma splice variants interacting partners, intracellular targeting, and regulation will be defined. The roles played by these PIPKIgamma isoforms in cell migration will be investigated. (3) The role of PIPKIgamma splice isoforms in breast cancer intravasation and metastasis will be defined using a mouse model. (4) Changes in PIPKIgamma content and isoform expression in breast tumors will be studied using a large and well characterized breast tumor tissue microarray. We will investigate changes in PIPKIgamma expression levels in breast tumors and correlate this with other signaling molecules and biomarkers and with patient outcomes.
This Aim will be the beginning of a long-term study to relate changes in PIPKIgamma expression with tumors of epithelial origin. This approach may yield important information, which will help to define the underlying mechanisms for our cell and molecular biology studies. In addition, our mechanistic studies will be translated into a greater understanding of breast cancer cell invasiveness.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM057549-12
Application #
7821343
Study Section
Tumor Progression and Metastasis Study Section (TPM)
Program Officer
Jones, Warren
Project Start
1998-05-01
Project End
2011-04-30
Budget Start
2010-05-01
Budget End
2011-04-30
Support Year
12
Fiscal Year
2010
Total Cost
$355,804
Indirect Cost
Name
University of Wisconsin Madison
Department
Pharmacology
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Tan, Xiaojun; Anderson, Richard A (2017) Keeping in touch with the ER network. Science 356:584-585
Choi, Suyong; Anderson, Richard A (2016) IQGAP1 is a phosphoinositide effector and kinase scaffold. Adv Biol Regul 60:29-35
Thapa, Narendra; Tan, Xiaojun; Choi, Suyong et al. (2016) The Hidden Conundrum of Phosphoinositide Signaling in Cancer. Trends Cancer 2:378-390
Choi, Suyong; Hedman, Andrew C; Sayedyahossein, Samar et al. (2016) Agonist-stimulated phosphatidylinositol-3,4,5-trisphosphate generation by scaffolded phosphoinositide kinases. Nat Cell Biol 18:1324-1335
Tan, Xiaojun; Thapa, Narendra; Liao, Yihan et al. (2016) PtdIns(4,5)P2 signaling regulates ATG14 and autophagy. Proc Natl Acad Sci U S A 113:10896-901
Tan, Xiaojun; Lambert, Paul F; Rapraeger, Alan C et al. (2016) Stress-Induced EGFR Trafficking: Mechanisms, Functions, and Therapeutic Implications. Trends Cell Biol 26:352-366
Thapa, Narendra; Choi, Suyong; Tan, Xiaojun et al. (2015) Phosphatidylinositol Phosphate 5-Kinase I? and Phosphoinositide 3-Kinase/Akt Signaling Couple to Promote Oncogenic Growth. J Biol Chem 290:18843-54
Tan, Xiaojun; Thapa, Narendra; Sun, Yue et al. (2015) A kinase-independent role for EGF receptor in autophagy initiation. Cell 160:145-60
Choi, Suyong; Thapa, Narendra; Tan, Xiaojun et al. (2015) PIP kinases define PI4,5P?signaling specificity by association with effectors. Biochim Biophys Acta 1851:711-23
Tan, Xiaojun; Thapa, Narendra; Choi, Suyong et al. (2015) Emerging roles of PtdIns(4,5)P2--beyond the plasma membrane. J Cell Sci 128:4047-56

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