Laminin (LM) is a large heterotrimeric extracellular matrix glycoprotein that plays important roles in cell adhesion, proliferation, migration, contraction, and differentiation. Each glomerular cell type expresses a unique subset of LM isoforms. Temporal switches in their expression correlate with morphologic transitions during glomerulogenesis. In some cases, initiation of cellular differentiation or maintenance of cellular pheontype has been linked to a particular LM isoform. Glomerular mesangial cells (MC) express LM-8 (a1B1g1), which binds entactin, cross-links into the fibrillar extracellular matrix and facilitates attachment and spreading. MC also express LM-9 (a1B1g1), which remains localized to discrete regions on the cell surface and initiates filopodia formation. This proposal will further characterize the mechanisms whereby LM isoforms direct MC function. In the first specific aim, we propose that the LMa4 chain forms a complex with filamin-1 (actin binding protein 280) that functions to generate force and mechano-protection of MC during contraction. Because this complex is shared with vascular smooth muscle cells and smooth muscle in the intestine, we propose that it is the smooth muscle equivalent of the LMa2-dystroglycan-dystrophin complex that is required for skeletal muscle contractility. The two LM isoforms expressed by MC differ in their use of B chains; thus it is likely that they also engage proteins on the MC surface that convey additional specificity of function. Proposed experiments will identify the molecular partners in these complexes. The goal of the second specific aim is to define the LM-receptors complexes that mediate MC contraction and migration. During biologic responses to a variety of stimuli, proteases are released that generate specific fragments of LM. In some cases these fragments acquire new biologic activity. Preliminary data show that a fragment of the LM B1 chain is translocated to the nucleus where it regulates gene expression. It is the goal of the third specific aim to confirm these observations and elucidate mechanisms whereby LMB1 acts in the nucleus. Progressive glomerular diseases such as glomerulosclerosis of aging and diabetic nephropathy are characterized by changes in the expression of LM isoforms. Further elucidation of the mechanisms whereby LM isoforms modulate the MC capacity to contract, protect the capillary wall from over distention, migrate during wound healing, and maintain normal cellular phentype and function should provide important insights into disease pathogenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK049771-08
Application #
6607443
Study Section
Pathology A Study Section (PTHA)
Program Officer
Ketchum, Christian J
Project Start
1996-05-01
Project End
2006-07-31
Budget Start
2003-08-01
Budget End
2006-07-31
Support Year
8
Fiscal Year
2003
Total Cost
$216,720
Indirect Cost
Name
University of Washington
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Abrass, Christine K; Hansen, Kim M; Patton, Bruce L (2010) Laminin alpha4-null mutant mice develop chronic kidney disease with persistent overexpression of platelet-derived growth factor. Am J Pathol 176:839-49
Abrass, Christine K; Hansen, Kim M (2010) Insulin-like growth factor-binding protein-5-induced laminin gamma1 transcription requires filamin A. J Biol Chem 285:12925-34
Schaeffer, Valerie; Hansen, Kim M; Morris, David R et al. (2010) Reductions in laminin beta2 mRNA translation are responsible for impaired IGFBP-5-mediated mesangial cell migration in the presence of high glucose. Am J Physiol Renal Physiol 298:F314-22
Zager, Richard A; Johnson, Ali C; Lund, Steve et al. (2006) Levosimendan protects against experimental endotoxemic acute renal failure. Am J Physiol Renal Physiol 290:F1453-62
Hansen, Kim; Abrass, Christine K (2003) Laminin-8/9 is synthesized by rat glomerular mesangial cells and is required for PDGF-induced mesangial cell migration. Kidney Int 64:110-8
Berfield, Anne K; Andress, Dennis L; Abrass, Christine K (2002) IGF-1-induced lipid accumulation impairs mesangial cell migration and contractile function. Kidney Int 62:1229-37
Berfield, A K; Andress, D L; Abrass, C K (2000) IGFBP-5(201-218) stimulates Cdc42GAP aggregation and filopodia formationin migrating mesangial cells. Kidney Int 57:1991-2003
Hansen, K; Abrass, C K (1999) Role of laminin isoforms in glomerular structure. Pathobiology 67:84-91
Hansen, K M; Berfield, A K; Spicer, D et al. (1998) Rat mesangial cells express two unique isoforms of laminin which modulate mesangial cell phenotype. Matrix Biol 17:117-30
Abrass, C K; Spicer, D; Berfield, A K et al. (1997) Diabetes induces changes in glomerular development and laminin-beta 2 (s-laminin) expression. Am J Pathol 151:1131-40