Our long-term interest continues to be understanding adhesive interactions between epithelial cells and basement membranes, which regulate epithelial tissue formation and maintenance. Laminin-5 (Ln-5) is a unique basement membrane ligand for epithelial cells. We and others determined that no other extracellular matrix was as effective as Ln-5 in promoting efficient attachment and spreading of epithelial cells. However, behavior of cells in contact with Ln-5 may vary from static adhesion and hemidesmosome formation, to vigorous migration. We hypothsize that these apparently opposing functions of Ln-5 in vitro are the reflection of regulatory mechanisms underlying epithelial morphogenesis in vivo. The goal of this proposal is to elucidate these mechanisms. We hypothesize tht regultion of spreading, static adhesion and migration occurs at several levels, including structure/ function modulation of Ln-5 and of its receptors and modifications in the components of the focal complexes in contact with Ln-5. We are in an excellent position to study these mechanisms, since we have identified a source for purified rat Ln-5 protein, we are advanced in the characterization of integrins that bind Ln-5, and we have defined a system of paried cell types with either static or migratory behavior on ln-5. We will: 1) complete the structural and functional characterization of Ln-5, by a combination of recombinant DNA techniques, purification of intact or fragmented protein and functional epitope mapping; 2) thoroughly characterize its receptors, especially integrins, by binding and competition assays; 3) define and test a mechanistic model for the regulation of adhesion and migration on Ln- 5, using a series of assays measuring strength of adhesion, spreading, migration and motility in a system of paried cell lines; 4) define the regulatory role of molecular components of focal complexes by cDNA cloning, mutagenesis, transfection and testing in the cell behavior assays. By these approaches, we hope to better our understanding of molecular interactions occuring in the focal complexes of epithelial cells in contact with Ln-5, and to help elucidate the molecular mechanisms of cell spreading, static adhesion and migration on Ln-5, and their hierarchy in the context of epithelial tissue morphogenetic events. Clarifying mechanical and biochemical aspects of epithelial adhesion to basement membranes may engender a better understanding of human diseases caused by epithelial tissue degeneration, including cancer metastasis, and lay the foundations for rational approaches to tissue healing and regeneration.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
Project #
Application #
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Scripps Research Institute
La Jolla
United States
Zip Code
Hintermann, Edith; Yang, Neng; O'Sullivan, Deirdre et al. (2005) Integrin alpha6beta4-erbB2 complex inhibits haptotaxis by up-regulating E-cadherin cell-cell junctions in keratinocytes. J Biol Chem 280:8004-15
Koshikawa, Naohiko; Minegishi, Tomoko; Sharabi, Andrew et al. (2005) Membrane-type matrix metalloproteinase-1 (MT1-MMP) is a processing enzyme for human laminin gamma 2 chain. J Biol Chem 280:88-93
Bilban, Martin; Ghaffari-Tabrizi, Nassim; Hintermann, Edith et al. (2004) Kisspeptin-10, a KiSS-1/metastin-derived decapeptide, is a physiological invasion inhibitor of primary human trophoblasts. J Cell Sci 117:1319-28
Schenk, Susann; Hintermann, Edith; Bilban, Martin et al. (2003) Binding to EGF receptor of a laminin-5 EGF-like fragment liberated during MMP-dependent mammary gland involution. J Cell Biol 161:197-209
Hendrix, Mary J C; Seftor, Elisabeth A; Kirschmann, Dawn A et al. (2003) Remodeling of the microenvironment by aggressive melanoma tumor cells. Ann N Y Acad Sci 995:151-61
Bilban, Martin; Buehler, Lukas K; Head, Steven et al. (2002) Normalizing DNA microarray data. Curr Issues Mol Biol 4:57-64
Hintermann, Edith; Haake, Susan Kinder; Christen, Urs et al. (2002) Discrete proteolysis of focal contact and adherens junction components in Porphyromonas gingivalis-infected oral keratinocytes: a strategy for cell adhesion and migration disabling. Infect Immun 70:5846-56
Seftor, R E; Seftor, E A; Koshikawa, N et al. (2001) Cooperative interactions of laminin 5 gamma2 chain, matrix metalloproteinase-2, and membrane type-1-matrix/metalloproteinase are required for mimicry of embryonic vasculogenesis by aggressive melanoma. Cancer Res 61:6322-7
Hintermann, E; Bilban, M; Sharabi, A et al. (2001) Inhibitory role of alpha 6 beta 4-associated erbB-2 and phosphoinositide 3-kinase in keratinocyte haptotactic migration dependent on alpha 3 beta 1 integrin. J Cell Biol 153:465-78
Kiosses, W B; Hahn, K M; Giannelli, G et al. (2001) Characterization of morphological and cytoskeletal changes in MCF10A breast epithelial cells plated on laminin-5: comparison with breast cancer cell line MCF7. Cell Commun Adhes 8:29-44

Showing the most recent 10 out of 28 publications