Two complementary approaches have been used to provide insight into the molecular mechanisms of adhesion of human or rodent neuroblastoma tumor cells to a model fibronectin-containing extracellular matrix--(a) biochemical analyses of the components in substratum adhesion sites after EGTA-mediated detachment of cells from the tissue culture substratum and (b) evaluation of adhesive responses of neural tumor cells to substrata coated with specific binding domains of the fibronectin moleculle or model proteins with specific binding activities. These studies reveal that (a) the mechanisms of adhesion of the growth cone and the cell body of neuroblastoma cells are distinctly different, (b) neuroblastoma tumor cells from the central nervous system of the animal resond to defined matrices very differently than peripheral neurons and possible CNS neurons indicating strong selection for a particular matrix-adhering type of tumor cell, and (c) heparan sulfate proteoglycan on the cell surface plays a central role in cell body adhesion of neuroblastoma whereas the unidentified receptor for fibronectin plays a critical role in growth cone adhesion. In addition, neurite-containing neuroblastoma cells display on their surfaces and in their substratum adhesion sites a unique and very high molecular weight glycoconjugate, called Io, that is not observed on a host of other cells, including glioma tumor cells and nonneurite neuroblastoma.
Three specific aims will now be pursued. A more detailed investigation of the heparan sulfate proteoglycan in the substratum adhesion sites of neuroblastoma cells will be undertaken, including their aggregation competence and ability to bind to affinity matrices for correlation with adhesion functional analyses. Secondly, the structure of the unique Io glycoconjugate will be investigated based on preliminary hints as to its composition; included here is the raising of antibodies for both structural and cell biological studies of this neurite-""""""""associated"""""""" molecule. And, finally, adhesive responses to specific binding domains of fibronectin or laminin, as well as model binding proteins, will be used to define the receptor-dependent processes for growth cone or cell body adhesion of these neural tumor cells. These experiments will define similarities and differences in matrix adhesion of these tumor cells in concert with parallel studies with embryonic peripheral or central nervous system neurons, as well as the nature of the selection processes that give rise to tumor cells competent for penetrating fibronectin- and/or laminin-containing matrices.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS017139-06
Application #
3397391
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1981-04-01
Project End
1991-03-31
Budget Start
1986-04-01
Budget End
1987-03-31
Support Year
6
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Case Western Reserve University
Department
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
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Culp, L A; Lin, W; Kleinman, N R et al. (1998) Earliest steps in primary tumor formation and micrometastasis resolved with histochemical markers of gene-tagged tumor cells. J Histochem Cytochem 46:557-68
Judware, R; Culp, L A (1997) N-myc over-expression downregulates alpha3beta1 integrin expression in human Saos-2 osteosarcoma cells. Clin Exp Metastasis 15:228-38
Judware, R; Culp, L A (1997) Concomitant down-regulation of expression of integrin subunits by N-myc in human neuroblastoma cells: differential regulation of alpha2, alpha3 and beta1. Oncogene 14:1341-50
Judware, R; Culp, L A (1995) Over-expression of transfected N-myc oncogene in human SKNSH neuroblastoma cells down-regulates expression of beta 1 integrin subunit. Oncogene 11:2599-607
Judware, R; Lechner, R; Culp, L A (1995) Inverse expressions of the N-myc oncogene and beta 1 integrin in human neuroblastoma: relationships to disease progression in a nude mouse model system. Clin Exp Metastasis 13:123-33
Flickinger, K S; Judware, R; Lechner, R et al. (1994) Integrin expression in human neuroblastoma cells with or without N-myc amplification and in ectopic/orthotopic nude mouse tumors. Exp Cell Res 213:156-63
Kleinman, N R; Lewandowska, K; Culp, L A (1994) Tumour progression of human neuroblastoma cells tagged with a lacZ marker gene: earliest events at ectopic injection sites. Br J Cancer 69:670-9
Lewandowska, K; Pergament, E; Sukenik, C N et al. (1992) Cell-type-specific adhesion mechanisms mediated by fibronectin adsorbed to chemically derivatized substrata. J Biomed Mater Res 26:1343-63

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