Whereas it is well established that microvascular cells play crucial roles in the etiology of diabetic retinopathy and retinopathy of prematurity, the molecular and cellular mechanisms regulating disease inception remain obscure. To approach the mechanisms regulating these retinal vasoproliferative disorders, antibody and recombinant DNA probes will be used in model systems. Results will reveal the role that cell-matrix, growth factor-matrix and cell-cell interactions play in retinal microvascular differentiation and pathology. Retinal microvascular endothelial cells and pericytes isolated from developing and post-natal retinas will be cultured alone and together. Rigid or mechanically deformable substrates, which have been synthesized by retinal vascular cells and previously shown by our group to differentially regulate vascular cell growth and contractile phenotype, will be assessed. Specifically, we will characterize the molecular mechanisms regulating retinal pericyte development and differentiation as well as the retinal microvascular endothelial cell response to injury. To this end, we will use the combined approaches of: (i) antibody localization using contractile protein isoform-specific and growth factor-specific antibodies prepared and characterized in the lab, (ii) quantitative immunoprecipitation and fluorographic analysis of biolabeled subcellular fractions derived from developing, differentiated and injured cell populations, (iii) Northern blot and transcription run-off analyses of injured endothelial cells, growth-arrested and growth-stimulated pericytes purified from neonatal and adult retinas and (iv) PCR, homology probing and antibody screening of lambda Zap II cDNA libraries derived from retinal pericytes using probes designed to reveal genetic elements regulating retinal microvascular myogenic determination. Results of these interdisciplinary studies will help to determine whether growth factor-matrix, matrix-cell and/or cell-cell interactions modulate the expression of cis DNA regulatory elements and trans-acting factors responsible for controlling retinal microvascular growth, differentiation and the unmanageable vasoproliferation associated with diabetic retinopathy.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
1R01EY009033-01A2
Application #
3266364
Study Section
Visual Sciences C Study Section (VISC)
Project Start
1992-12-01
Project End
1996-11-30
Budget Start
1992-12-01
Budget End
1993-11-30
Support Year
1
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Tufts University
Department
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02111
Demidova-Rice, Tatiana N; Geevarghese, Anita; Herman, Ira M (2011) Bioactive peptides derived from vascular endothelial cell extracellular matrices promote microvascular morphogenesis and wound healing in vitro. Wound Repair Regen 19:59-70
Durham, Jennifer T; Herman, Ira M (2009) Inhibition of angiogenesis in vitro: a central role for beta-actin dependent cytoskeletal remodeling. Microvasc Res 77:281-8
Kutcher, Matthew E; Herman, Ira M (2009) The pericyte: cellular regulator of microvascular blood flow. Microvasc Res 77:235-46
Herman, Ira M; Leung, Alice (2009) Creation of human skin equivalents for the in vitro study of angiogenesis in wound healing. Methods Mol Biol 467:241-8
Kutcher, Matthew E; Kolyada, Alexey Y; Surks, Howard K et al. (2007) Pericyte Rho GTPase mediates both pericyte contractile phenotype and capillary endothelial growth state. Am J Pathol 171:693-701
Sieczkiewicz, Gregory J; Herman, Ira M (2003) TGF-beta 1 signaling controls retinal pericyte contractile protein expression. Microvasc Res 66:190-6
Potter, David A; Srirangam, Anjaiah; Fiacco, Kerry A et al. (2003) Calpain regulates enterocyte brush border actin assembly and pathogenic Escherichia coli-mediated effacement. J Biol Chem 278:30403-12
Papetti, Michael; Shujath, Jaleel; Riley, Kathleen N et al. (2003) FGF-2 antagonizes the TGF-beta1-mediated induction of pericyte alpha-smooth muscle actin expression: a role for myf-5 and Smad-mediated signaling pathways. Invest Ophthalmol Vis Sci 44:4994-5005
Kolyada, Alexey Y; Riley, Kathleen N; Herman, Ira M (2003) Rho GTPase signaling modulates cell shape and contractile phenotype in an isoactin-specific manner. Am J Physiol Cell Physiol 285:C1116-21
Riley, Kathleen N; Maldonado, Angel E; Tellier, Patrice et al. (2003) Betacap73-ARF6 interactions modulate cell shape and motility after injury in vitro. Mol Biol Cell 14:4155-61

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