Deconstructing Directional Cell Motility in Metastasis thru Substratum nanopatter

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The goal of this proposal is the development of anti-metastasis therapeutics. To achieve this goal, we propose a novel, multidisciplinary approach that combines nano-technology with molecular and cell biology. The rationale is to deconstruct the directional cell motility mechanisms that underlie invasiveness through control of cell geometry. The premise of the approach is that control of cell geometry will allow us to elicit stereotypical structural and functional cell phenotypes characteristic of defined phases of the motility cycle. Arrays of essentially identical "designer cells" will provide a unique platform for probing complex directional motility mechanisms and for establishing high-content, translational screening procedures. We will first develop the nano-technology to control the geometry and, therefore, the motility machinery of cells. Technologies will be developed that are compatible with live cell imaging and electron microscopy. Novel technologies will include wet stamping (WETS) to create transparent adhesive islands, controlled etching (GET) to generate nano-patterns from micro-patterns and parallel array dip pen lithography (DPN) to direct write nano-features. Based on this technology, we will then generate patterned substrata, including circles, triangles, tear-drops and polygons surrounded by satellite nano-spots to deconstruct directional motility mechanisms. We will focus on focal adhesion (FA)-microtubule (MT) interaction and focal adhesion turnover; cell polarization and lamellipodial protrusion; filopodial formation and MT interaction; and membrane protease recruitment. Gene silencing in combination with high-content screening of arrays of identical, "designer" cells will lead to identification of molecular targets and pathways crucial for directional motility. These targets and pathways will then serve as the basis for screens to identify small molecule inhibitors of metastasis.