DF/HCC Program Affiliation

Member, Invasion, Metastasis, and Angiogenesis Program

Research Abstract

Cell crawling behavior is essential for human development, maintenance and defense, and instrumental in disease processes such as inflammation and the spread of cancer cells. The research concerns the machinery used by human white blood cells, tissue defense cells, tumor cells and blood platelets to crawl and change shape respectively. It focuses on two general questions based on key cell protein components which control how the major cell protein, actin, forms struts and levers that control cell shape and movements. These actin-regulating components were isolated originally from lung defense cells, the alveolar macrophages, and are believed important for lung protection and disease. The characterization of these proteins led to principles believed to explain how actin architecture, the so-called actin cytoskeleton, is maintained in the cell and how this architecture is changed to accommodate crawling movements. We set out to explain how one of the components, a protein called filamin-A, causes actin filaments to take on particular configurations within the cell and how signaling processes that mediate instructions delivered from outside cells to elicit crawling behavior might regulate filamin-A's functions, which also include linking the actin cytoskeleton to plasma membrane receptors and serving as a scaffold for cellular trafficking and signaling reactions. The proposal also plans to examine how human blood neutrophils, the most rapidly crawling of our cells, determine where and when to assemble new actin filaments, based on hypotheses derived from a systematic study of the second component, a protein named gelsolin. The investigations involve elucidation of how signal intermediates such as GTPases and phospholipids work to promote actin polymerization. The eventual goal is to understand these processes sufficiently to modify them in hopes of mollifying inflammation and metastatic tumor spread. One practical spinoffs of this basic research includes the discoveryies that a circulating form of gelsolin is an anti-inflammatory component of the blood and that its depletion in states of injury and inflammation predict secondary injury such as acute lung injury and the adult respiratory distess syndrome. We propose that gelsolin replacement could prevent such secondary injury. A second spinoff is the development of a technology permitting the storage of blood platelets under refrigeration, something currently not possible. If successful this technology could have major impacts on transfusion medicine and the support of cancer patients.

Publications

• Di Nardo A, Cicchetti G, Falet H, Hartwig JH, Stossel TP, Kwiatkowski DJ. Arp2/3 complex-deficient mouse fibroblasts are viable and have normal leading-edge actin structure and function. Proc Natl Acad Sci U S A 2005 Nov 8; 102(45):16263-8
  PMID: 16254049
• Stossel TP. Regulating academic-industrial research relationships--solving problems or stifling progress? N Engl J Med 2005 Sep 8; 353(10):1060-5
  PMID: 16148294
• Nakamura F, Hartwig JH, Stossel TP, Szymanski PT. Ca2+ and calmodulin regulate the binding of filamin A to actin filaments. J Biol Chem 2005 Sep 16; 280(37):32426-33
  PMID: 16030015
• Josefsson EC, Gebhard HH, Stossel TP, Hartwig JH, Hoffmeister KM. The macrophage alphaMbeta2 integrin alphaM lectin domain mediates the phagocytosis of chilled platelets. J Biol Chem 2005 May 6; 280(18):18025-32
  PMID: 15741160
• Valeri CR, Ragno G, Marks PW, Kuter DJ, Rosenberg RD, Stossel TP. Effect of thrombopoietin alone and a combination of cytochalasin B and ethylene glycol bis(beta-aminoethyl ether) N,N'-tetraacetic acid-AM on the survival and function of autologous baboon platelets stored at 4 degrees C for as long as 5 days. Transfusion 2004 Jun; 44(6):865-70
  PMID: 15157253
• Woo MS, Ohta Y, Rabinovitz I, Stossel TP, Blenis J. Ribosomal S6 kinase (RSK) regulates phosphorylation of filamin A on an important regulatory site. Mol Cell Biol 2004 Apr; 24(7):3025-35
  PMID: 15024089
• Hoffmeister KM, Josefsson EC, Isaac NA, Clausen H, Hartwig JH, Stossel TP. Glycosylation restores survival of chilled blood platelets. Science 2003 Sep 12; 301(5639):1531-4
  PMID: 12970565
• Hoffmeister KM, Felbinger TW, Falet H, Denis CV, Bergmeier W, Mayadas TN, von Andrian UH, Wagner DD, Stossel TP, Hartwig JH. The clearance mechanism of chilled blood platelets. Cell 2003 Jan 10; 112(1):87-97
  PMID: 12526796
• Christofidou-Solomidou M, Scherpereel A, Solomides CC, Christie JD, Stossel TP, Goelz S, DiNubile MJ. Recombinant plasma gelsolin diminishes the acute inflammatory response to hyperoxia in mice. J Investig Med 2002 Jan; 50(1):54-60
  PMID: 11813829
• Nakamura F, Osborn E, Janmey PA, Stossel TP. Comparison of filamin A-induced cross-linking and Arp2/3 complex-mediated branching on the mechanics of actin filaments. J Biol Chem 2002 Mar 15; 277(11):9148-54
  PMID: 11786548
• Stossel TP, Condeelis J, Cooley L, Hartwig JH, Noegel A, Schleicher M, Shapiro SS. Filamins as integrators of cell mechanics and signalling. Nat Rev Mol Cell Biol 2001 Feb; 2(2):138-45
  PMID: 11252955
• Stossel TP, Hartwig JH, Janmey PA, Kwiatkowski DJ. Cell crawling two decades after Abercrombie. Biochem Soc Symp 1999; 65:267-80
  PMID: 10320944
• Ohta Y, Suzuki N, Nakamura S, Hartwig JH, Stossel TP. The small GTPase RalA targets filamin to induce filopodia. Proc Natl Acad Sci U S A 1999 Mar 2; 96(5):2122-8
  PMID: 10051605
• Azuma T, Witke W, Stossel TP, Hartwig JH, Kwiatkowski DJ. Gelsolin is a downstream effector of rac for fibroblast motility. EMBO J 1998 Mar 2; 17(5):1362-70
  PMID: 9482733
• Hartwig JH, Bokoch GM, Carpenter CL, Janmey PA, Taylor LA, Toker A, Stossel TP. Thrombin receptor ligation and activated Rac uncap actin filament barbed ends through phosphoinositide synthesis in permeabilized human platelets. Cell 1995 Aug 25; 82(4):643-53
  PMID: 7664343
• Witke W, Sharpe AH, Hartwig JH, Azuma T, Stossel TP, Kwiatkowski DJ. Hemostatic, inflammatory, and fibroblast responses are blunted in mice lacking gelsolin. Cell 1995 Apr 7; 81(1):41-51
  PMID: 7720072
• Stossel TP. The machinery of cell crawling. Sci Am 1994 Sep; 271(3):54-5, 58-63
  PMID: 8091190