Pancreatic cancer is a devastating disease with an exceptionally poor prognosis. Tumors of ductular origin comprise 95% of pancreatic neoplasms and have strong tendencies to invade surrounding parenchyma and metastasize to distal organs. Currently, the molecular mechanisms that support these critical processes are poorly defined. It is known that the actin cytoskeleton plays an important role and responds dynamically to stimulation by a family of Receptor Tyrosine Kinases (RTKs) that are upregulated significantly in many pancreatic tumors. This is a proposal to study the mechanisms by which RTKs activate dramatic reorganization of the actin cytoskeleton, which alters cell polarity and leads to cell migration, invasion, and metastasis. We have identified a key regulatory cytoskeletal complex in these cells composed of;a large mechanochemical GTPase called dynamin (Dyn2) that is markedly upregulated in human pancreatic tumors, the actin crosslinking protein cortactin, and the transforming kinase Src, which is activated by RTKs and binds and phosphorylates both Dyn2 and cortactin. Importantly, we have shown that this complex associates with the actin cytoskeleton, cellular membranes, as well as known oncogenic proteins implicated in pancreatic cancer to form "signaling platforms" that support the invasive process. Our recent manuscripts and unpublished observations provide strong support for the CENTRAL HYPOTHESIS of this study: mainly, the Dyn2-Cort-Src complex, in concert with the oncogenic proteins ?-actinin and Rac, is activated by elevated RTK cascades to mediate actin-plasma membrane dynamics that promote invasion and metastasis of neoplastic pancreatic ductular cells. This proposal will utilize state-of-the-art optical and molecular methods applied to pancreatic tumor cell lines, tissue micro-array (TMA) technology of human tumor samples, and in vivo animal models to define the molecular mechanisms by which this protein complex potentiates pancreatic neoplasia and metastasis. This proposal is organized into three distinct but related specific aims that will define the following; first, how these signaling platforms mediate large-scale internalization of RTKs from the cell surface to alter cell signaling cascades;second, how the Dyn2-Cort-Src complex mediates major actin reorganization in tumor cells that become migratory and invasive;and third, how tumor cell adhesion complexes form and proliferate to mediate degradation of the surrounding extracellular matrix to facilitate metastasis. This study will make close correlations between isolated cells and human tumors to provide novel information toward understanding the molecular mechanisms that make pancreatic tumors so aggressive, migratory, and lethal. In addition, it describes pre-clinical trials using novel, recently identified pharmacological inhibitors.

Public Health Relevance

Pancreatic cancers are particularly lethal due to the aggressive way they spread and metastasize. This proposal will define some of the central mechanisms by which these tumor cells move away from the pancreas to invade peripheral organs by focusing on specific ongogenic proteins that are markedly upregulated in these cancers.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA104125-09
Application #
8473810
Study Section
Clinical, Integrative and Molecular Gastroenterology Study Section (CIMG)
Program Officer
Ault, Grace S
Project Start
2003-12-01
Project End
2015-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
9
Fiscal Year
2013
Total Cost
$259,092
Indirect Cost
$94,798
Name
Mayo Clinic, Rochester
Department
Type
DUNS #
006471700
City
Rochester
State
MN
Country
United States
Zip Code
55905
Razidlo, Gina L; Schroeder, Barbara; Chen, Jing et al. (2014) Vav1 as a central regulator of invadopodia assembly. Curr Biol 24:86-93
McNiven, Mark A (2013) Breaking away: matrix remodeling from the leading edge. Trends Cell Biol 23:16-21
Razidlo, Gina L; Wang, Yu; Chen, Jing et al. (2013) Dynamin 2 potentiates invasive migration of pancreatic tumor cells through stabilization of the Rac1 GEF Vav1. Dev Cell 24:573-85
Wang, Yu; McNiven, Mark A (2012) Invasive matrix degradation at focal adhesions occurs via protease recruitment by a FAK-p130Cas complex. J Cell Biol 196:375-85
Eppinga, R D; Krueger, E W; Weller, S G et al. (2012) Increased expression of the large GTPase dynamin 2 potentiates metastatic migration and invasion of pancreatic ductal carcinoma. Oncogene 31:1228-41
Feng, H; Liu, K W; Guo, P et al. (2012) Dynamin 2 mediates PDGFR*-SHP-2-promoted glioblastoma growth and invasion. Oncogene 31:2691-702
Schroeder, Barbara; Weller, Shaun G; Chen, Jing et al. (2010) A Dyn2-CIN85 complex mediates degradative traffic of the EGFR by regulation of late endosomal budding. EMBO J 29:3039-53
Weller, Shaun G; Capitani, Mirco; Cao, Hong et al. (2010) Src kinase regulates the integrity and function of the Golgi apparatus via activation of dynamin 2. Proc Natl Acad Sci U S A 107:5863-8
Orlichenko, Lidiya; Weller, Shaun G; Cao, Hong et al. (2009) Caveolae mediate growth factor-induced disassembly of adherens junctions to support tumor cell dissociation. Mol Biol Cell 20:4140-52
Kruchten, Anne E; Krueger, Eugene W; Wang, Yu et al. (2008) Distinct phospho-forms of cortactin differentially regulate actin polymerization and focal adhesions. Am J Physiol Cell Physiol 295:C1113-22

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