Cell migration and tumor cell invasion are modulated by integrin-mediated interactions between the extracellular matrix (ECM) and the actin cytoskeleton. Dynamic regulation of adhesions and reorganization of the actin cytoskeleton are required for cell migration and are critical determinants of the invasive properties of human tumors. In previous funding periods we established that regulated intracellular proteolysis by the calcium-dependent protease calpain2 is important for cell migration by targeting specific substrates to regulate the dynamic turnover of adhesions. We now propose to investigate how calpains and associated proteins including FAK and PCH family members modulate the temporal and spatial activation of Src kinases and specific substrate proteolysis to affect adhesion dynamics during invasive cell migration. Invasive cells frequently contain two types of cell matrix adhesions known as focal adhesions and podosome-type adhesions or invadopodia. Src kinases are critical for regulating the balance between the formation of focal adhesions and the generation of matrix-degrading invadopodia, found in invasive cancer cells. We have shown that FAK and calpain2 regulate the temporal and spatial activation of c-Src and mediate a balance between focal adhesion and invadopodia formation. The hypothesis that guides this research is that calpain2 associates in complexes with FAK, Src and PCH family members, that temporally and spatially modulate c-Src activity and substrate proteolysis to affect the formation and turnover of focal adhesions (FA) and invadopodia that mediate invasive cell migration. We are now uniquely positioned to make rapid progress with the following Specific Aims: 1. Investigate how FAK/calpain2/Src complex regulates the temporal and spatial dynamics of focal adhesions and invadopodia to affect cancer cell invasion. 2. Test the hypothesis that proteolysis of specific effector substrates by calpain2 regulates adhesion dynamics and cancer cell invasion. 3. Test the hypothesis that PCH proteins, PSTPIP1 and PACSIN2, associate with calpain2 and regulate the balance between focal adhesions and podosome-type adhesions to affect invasive cell migration. The research proposed in this grant will not only elucidate the molecular mechanisms that regulate adhesion dynamics during cell migration, but will also likely provide insight into the pathogenesis of invasive cancer cell migration and the development of new therapeutics for control of disease states such as tumor invasion and metastasis.

Public Health Relevance

The progression from a locally growing tumor to an invasive and metastatic tumor is the event that is most often responsible for treatment failures in patients with cancer. The proposed research contributes to human health by increasing our understanding of the molecular mechanisms that regulate cancer cell adhesion and invasive migration. Knowledge of the basic mechanisms that regulate cancer cell migration should aid in the identification of therapeutic targets that affect tumor invasion and metastasis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA085862-13
Application #
8212589
Study Section
Intercellular Interactions (ICI)
Program Officer
Woodhouse, Elizabeth
Project Start
2000-06-10
Project End
2015-02-28
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
13
Fiscal Year
2012
Total Cost
$278,257
Indirect Cost
$86,423
Name
University of Wisconsin Madison
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Starnes, Taylor W; Bennin, David A; Bing, Xinyu et al. (2014) The F-BAR protein PSTPIP1 controls extracellular matrix degradation and filopodia formation in macrophages. Blood 123:2703-14
Boateng, Lindsy R; Cortesio, Christa L; Huttenlocher, Anna (2012) Src-mediated phosphorylation of mammalian Abp1 (DBNL) regulates podosome rosette formation in transformed fibroblasts. J Cell Sci 125:1329-41
Cortesio, Christa L; Boateng, Lindsy R; Piazza, Timothy M et al. (2011) Calpain-mediated proteolysis of paxillin negatively regulates focal adhesion dynamics and cell migration. J Biol Chem 286:9998-10006
Huttenlocher, Anna; Horwitz, Alan Rick (2011) Integrins in cell migration. Cold Spring Harb Perspect Biol 3:a005074
Wernimont, Sarah A; Wiemer, Andrew J; Bennin, David A et al. (2011) Contact-dependent T cell activation and T cell stopping require talin1. J Immunol 187:6256-67
Chan, Keefe T; Bennin, David A; Huttenlocher, Anna (2010) Regulation of adhesion dynamics by calpain-mediated proteolysis of focal adhesion kinase (FAK). J Biol Chem 285:11418-26
Cortesio, Christa L; Perrin, Benjamin J; Bennin, David A et al. (2010) Actin-binding protein-1 interacts with WASp-interacting protein to regulate growth factor-induced dorsal ruffle formation. Mol Biol Cell 21:186-97
Wernimont, Sarah A; Legate, Kyle R; Simonson, William T N et al. (2010) PIPKI gamma 90 negatively regulates LFA-1-mediated adhesion and activation in antigen-induced CD4+ T cells. J Immunol 185:4714-23
Cortesio, Christa L; Wernimont, Sarah A; Kastner, Daniel L et al. (2010) Impaired podosome formation and invasive migration of macrophages from patients with a PSTPIP1 mutation and PAPA syndrome. Arthritis Rheum 62:2556-8
Chan, Keefe T; Cortesio, Christa L; Huttenlocher, Anna (2009) FAK alters invadopodia and focal adhesion composition and dynamics to regulate breast cancer invasion. J Cell Biol 185:357-70

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