Jan and Kathryn Ver Hagen Professor of Translational Research
B.S., University of Minnesota, St. Paul
Ph.D., University of Minnesota, Minneapolis
Postdoctoral Research, Washington University, St. Louis
Postdoctoral Research, Univ. of North Carolina, Chapel Hill
4533 WIMR II
1111 Highland Ave
Madison, WI 53705
Laboratory for Cell and Molecular Biology
Co-PI, Laboratory for Optical and Computational Instrumentation
Affiliate Department of Biomedical Engineering
Co-Leader, Tumor Microenvironment Group, UW Carbone Cancer Center
Visit the Keely lab page
Integrin and small GTPase signaling events in differentiation and transformation
Appropriate cellular interactions with the extracellular matrix (ECM)
help to establish normal cellular architecture and differentiation.
During oncogenic transformation, these normal interactions with the ECM are profoundly altered,
resulting in cells that lose their polarization and differentiation, lose anchorage dependent growth control, and
acquire a migratory, invasive phenotype.
Our lab is interested in understanding, at a molecular level, how cellular interactions with the ECM determine
differentiation and epithelial polarization, and how these interactions are altered during carcinogenesis to result in invasive, metastatic carcinoma.
Understanding molecular mechanisms underlying breast cancer risk due to breast density
Patients with "dense" breast tissue have a four to six-fold increased risk of developing breast carcinomas.
In fact, 1/3 of all breast cancer cases are attributed to breast density, making it one of the greatest risk factors
for carcinoma. Increased breast density is associated with a significant increase in the deposition of connective tissue,
or extracellular matrix (ECM) components, most notably the protein, collagen.
We have been developing model systems to understand why increased breast density results in an increased risk
for developing breast carcinoma.
We find in a simple in vitro model that increasing the density of collagen in the matrix is sufficient to
disrupt breast epithelial differentiation, suggesting that matrix density is itself an important regulator of cellular behavior.
Additionally, we are employing a mouse strain engineered to have more collagen in its connective tissue.
Mouse tumor studies are underway to directly test whether increased collagen density will enhance tumor formation or tumor metastasis.
State of the art imaging approaches are being used to characterize the collagen structure in
normal glands and around tumors so that we can better understand the physical relationship between
cells and the collagen fibers found in breast tissue.
We find evidence for a collagen "signature" that is present in even before a tumor is palpable, predicting
where a tumor will soon arise. We are investigating whether this signature can
be developed as a tool to aid in diagnosing human breast carcinoma at an earlier stage.
Using biochemical and DNA microarray approaches, we are characterizing several
biochemical and genetic changes that occur in cells that encounter dense matrices.
Dense collagen environments activate signal pathways within cells that result in a more
tumor-like behavior: increased cell proliferation or growth, decreased cell death, and increased ability to
invade into nearby tissues.
We expect that our studies to characterize the molecular response of cells to dense collagen matrices will allow us to better understand tumor progression.
Molecular signaling events related to cell interactions with the ECM
Cells interact with the ECM through a variety of cell surface receptors,
the best understood of which are members of the integrin family.
Much remains to be determined regarding the specific molecular players and signaling pathways
downstream of integrins, and how these pathways are involved in the progression of various diseases.
Therefore, part of the focus of the lab is to investigate signaling events through the integrin family of receptors.
A second aspect of this work is to investigate how small GTPases of the Ras superfamily,
some of which are known or suspected oncogenes, affect the response of cells to the ECM.
Specifically, we have focused on R-Ras and Rho,
which we find alter the way breast epithelial cells respond to the ECM, promoting cellular migration and invasion.
We are particularly interested in studying signaling events using state of the art imaging approaches
to understand how small GTPases function in a spatial and temporal manner during cell migration.
- Conklin, M.W., A. Ada-Nguema, M. Parsons, K.M. Riching, and P.J. Keely. 2010. R-Ras regulates beta1-integrin trafficking via effects on membrane ruffling and endocytosis. BMC Cell Biol. 11:14
- Sung, K.E., G. Su, C. Pehlke, S.M. Trier, K.W. Eliceiri, P.J. Keely, A. Friedl, and D.J. Beebe. 2009. Control of 3-dimensional collagen matrix polymerization for reproducible human mammary fibroblast cell culture in microfluidic devices. Biomaterials. 30:4833-41.
- Provenzano, P.P., and P.J. Keely. 2009. The role of focal adhesion kinase in tumor initiation and progression. Cell Adh Migr. 3.
- Provenzano, P.P., D.R. Inman, K.W. Eliceiri, and P.J. Keely. 2009. Matrix density-induced mechanoregulation of breast cell phenotype, signaling and gene expression through a FAK-ERK linkage. Oncogene. 28:4326-43
- Provenzano, P.P., K.W. Eliceiri, and P.J. Keely. 2009. Shining new light on 3D cell motility and the metastatic process. Trends Cell Biol. 19:638-48
- Provenzano, P.P., K.W. Eliceiri, and P.J. Keely. 2009. Multiphoton microscopy and fluorescence lifetime imaging microscopy (FLIM) to monitor metastasis and the tumor microenvironment. Clin Exp Metastasis. 26:357-70.
- Gehler, S., M. Baldassarre, Y. Lad, J.L. Leight, M.A. Wozniak, K.M. Riching, K.W. Eliceiri, V.M. Weaver, D.A. Calderwood, and P.J. Keely. 2009. Filamin A-beta1 integrin complex tunes epithelial cell response to matrix tension. Mol Biol Cell. 20:3224-38.
- Conklin, M.W., P.P. Provenzano, K.W. Eliceiri, R. Sullivan, and P.J. Keely. 2009. Fluorescence lifetime imaging of endogenous fluorophores in histopathology sections reveals differences between normal and tumor epithelium in carcinoma in situ of the breast. Cell Biochem Biophys. 53:145-57
- Provenzano, P.P., C.T. Rueden, S.M. Trier, L. Yan, S.M. Ponik, D.R. Inman, P.J. Keely, and K.W. Eliceiri. 2008. Nonlinear optical imaging and spectral-lifetime computational analysis of endogenous and exogenous fluorophores in breast cancer. J Biomed Opt. 13:031220
- Provenzano, P.P., D.R. Inman, K.W. Eliceiri, S.M. Trier, and P.J. Keely. 2008. Contact Guidance Mediated Three-Dimensional Cell Migration is Regulated by Rho/ROCK-Dependent Matrix Reorganization. Biophys J. 95:5374-5384
- Provenzano, P.P., D.R. Inman, K.W. Eliceiri, J.G. Knittel, L. Yan, C.T. Rueden, J.G. White, and P.J. Keely. 2008. Collagen density promotes mammary tumor initiation and progression. BMC Med. 6:11
- Provenzano, P.P., D.R. Inman, K.W. Eliceiri, H.E. Beggs, and P.J. Keely. 2008. Mammary epithelial-specific disruption of focal adhesion kinase retards tumor formation and metastasis in a transgenic mouse model of human breast cancer. Am J Pathol. 173:1551-65
- Provenzano, P.P., K.W. Eliceiri, L. Yan, A. Ada-Nguema, M.W. Conklin, D.R. Inman, and P.J. Keely. 2008. Nonlinear optical imaging of cellular processes in breast cancer. Microsc Microanal. 14:532-48
- Keely PJ, Conklin MW, Gehler S, Ponik SM, and Provenzano PP. (2007). Investigating integrin regulation and signaling events in three-dimensional systems. Methods Enzymol. 426:27-45. PMID 17697878
- Skala MC, Riching KM, Bird DK, Gendron-Fitzpatrick A, Eickhoff J, Eliceiri KW, Keely PJ, and Ramanujam N. (2007). In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia. J Biomed Opt. 12:024014. PMID 17477729
- Provenzano PP, Eliceiri KW, Campbell JM, Inman DR, White JG, and Keely PJ. (2007). Collagen reorganization at the tumor-stromal interface facilitates local invasion. BMC Med. 4:38. PMID 17190588
- Modzelewska K, Newman LP, Desai R, Keely PJ. (2007). Ack1 mediates Cdc42-dependent cell migration and signaling to p130Cas. J Biol Chem. 281:37527-37535. PMID 17038317
- Pal S, Wu J, Murray JK, Gellman SH, Wozniak MA, Keely PJ, Boyer ME, Gomez TM, Hasso SM, Fallon JF, and Bresnick EH. (2006). An antiangiogenic neurokinin-B/thromboxane A2 regulatory axis. J Cell Biol. 174:1047-58. PMID 17000881
- Ada-Nguema AS, Xenias H, Sheetz MP, and Keely PJ. (2006). The small GTPase R-Ras regulates organization of actin and drives membrane protrusions through the activity of PLCepsilon. J Cell Sci. 119(Pt 7):1307-19. PMID 16537651
- Wozniak MA and Keely PJ. (2005). Use of three-dimensional collagen gels to study mechanotransduction in T47D breast epithelial cells. Biol Proced Online. 7:144-61. PMID 16299584
- Bird DK, Yan L, Vrotsos KM, Eliceiri KW, Vaughan EM, Keely PJ, White JG, and Ramanujam N. (2005). Metabolic mapping of MCF10A human breast cells via multiphoton fluorescence lifetime imaging of the coenzyme NADH. Cancer Res. 65:8766-73. PMID 16204046
- Wozniak MA, Kwong L, Chodniewicz D, Klemke R, and Keely PJ. (2005). R-Ras controls membrane protrusion and cell migration through the spatial regulation of Rac and Rho. Mol Biol Cell. 16:84-96. PMID 15525681
- Wozniak MA, Modzelewska K, Kwong L, and Keely PJ. (2004). Focal adhesion regulation of cell behavior. Biochemica Biiophys Acta. 1692:103-119. PMID 15246682
- Palmer GM, Keely PJ, Breslin TM, and Ramanujam N. (2003). Autofluorescence spectroscopy of normal and malignant human breast cell lines. Photochem and Photobiol. 78:462-469. PMID 14653577