Approximately 1,000,000 aortocoronary and peripheral vascular revascularizations are performed using autologous conduits. The leading cause of graft failure is the subsequent development of intimal hyperplasia. Intimal hyperplasia represents a response to injury that involves smooth muscle proliferation, migration, phenotypic modulation, and extracellular matrix (ECM) deposition. This proposal will develop a cell permeant peptide therapeutic to enhance graft patency by preventing the events that lead to intimal hyperplasia. A small heat shock protein, HSP27, is phosphorylated by a kinase cascade involving p38 map kinase and MAPKAP kinase II (MK2). Phosphorylated HSP27 is associated with the formation of actin stress fibers (myofibroblast phenotype) and enhanced smooth muscle migration. We have developed a cell permeant peptide that inhibits MK2. This peptide also inhibits stress fiber formation and ECM production.
The specific aims of this proposal are:
Specific aim #1 : Determine the effect of transducible peptides which inhibit the phosphorylation of HSP27 on smooth muscle physiology, morphology, and biochemistry: We will determine the effect of the novel MK2 inhibitor peptide on intact human vascular smooth muscle segments and cultured vascular smooth muscle cells.
Specific aim #2 : Determine the effect of optimized peptide mimetics on intimal hyperplasia. We will first determine the effect of the MK2 inhibitor on intimal hyperplasia in a human saphenous vein graft organ culture model. Subsequently, we will determine the effect of the mimetics in vivo in a rabbit carotid interposition model.
Specific aim #3 : Determine the molecular mechanisms by which phosphorylated HSP27 """"""""stabilizes"""""""" the actin cytoskeleton: We will use quantitative, high throughput mass spectrometry techniques to analyze the molecular associations of phosphorylated and nonphosphorylated HSP27. The goal of this project is to engineer biomolecules that enhance graft patency using protein transduction domains to directly introduce peptides into smooth muscle cells. Autologous conduits represent an ideal target for this therapeutic approach in that the graft can be treated ex vivo, thus providing an optimal environment for the delivery of engineered protein/peptide therapeutics. The molecules designed in this proposal represent novel therapeutics in that the usual targets of drug development (cell surface receptors and signaling cascades) are """"""""bypassed"""""""" and protein-protein interactions are stoichiometrically altered by changing the phosphorylation of downstream target effector proteins (HSP27).

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL070715-07
Application #
7799153
Study Section
Bioengineering, Technology and Surgical Sciences Study Section (BTSS)
Program Officer
Reid, Diane M
Project Start
2002-04-01
Project End
2012-01-31
Budget Start
2010-02-01
Budget End
2011-01-31
Support Year
7
Fiscal Year
2010
Total Cost
$420,575
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Surgery
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
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Cheung-Flynn, Joyce; Song, Jun; Voskresensky, Igor et al. (2017) Limiting Injury During Saphenous Vein Graft Preparation For Coronary Arterial Bypass Prevents Metabolic Decompensation. Sci Rep 7:14179
Boire, Timothy C; Balikov, Daniel A; Lee, Yunki et al. (2016) Biomaterial-Based Approaches to Address Vein Graft and Hemodialysis Access Failures. Macromol Rapid Commun 37:1860-1880
Osgood, Michael J; Sexton, Kevin; Voskresensky, Igor et al. (2016) Use of Brilliant Blue FCF during vein graft preparation inhibits intimal hyperplasia. J Vasc Surg 64:471-478
Wise, Eric S; Hocking, Kyle M; Luo, Weifeng et al. (2016) Traditional graft preparation decreases physiologic responses, diminishes viscoelasticity, and reduces cellular viability of the conduit: A porcine saphenous vein model. Vasc Med 21:413-421
Hocking, Kyle M; Putumbaka, Gowthami; Wise, Eric S et al. (2016) Papaverine Prevents Vasospasm by Regulation of Myosin Light Chain Phosphorylation and Actin Polymerization in Human Saphenous Vein. PLoS One 11:e0154460
Hocking, Kyle M; Luo, Weifeng; Li, Fan Dong et al. (2016) Brilliant blue FCF is a nontoxic dye for saphenous vein graft marking that abrogates response to injury. J Vasc Surg 64:210-8
Wise, Eric S; Hocking, Kyle M; Feldman, Daniel et al. (2015) An Optimized Preparation Technique for Saphenous Vein Graft. Am Surg 81:E274-6
Wise, Eric S; Hocking, Kyle M; Eagle, Susan et al. (2015) Preservation solution impacts physiologic function and cellular viability of human saphenous vein graft. Surgery 158:537-46
Li, Fan Dong; Eagle, Susan; Brophy, Colleen et al. (2014) Pressure control during preparation of saphenous veins. JAMA Surg 149:655-62

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