Smooth muscle proliferation is an important feature of diseases such as atherosclerosis and hypertension, and is generally held responsible for restenosis after angioplasty and the failure of coronary bypass grafts. Associated with this process is an increased expression of nonmuscle myosin isoforms. The significance of this observation is presently unclear but our hypothesis is that this protein is required for cell division and that its deregulated expression is key to the abberrant proliferation seen in restenotic lesions. The long-term aim of this project is to understand the role of nonmuscle myosin in proliferating cells both in normal and disease states. We intend to determine the factors which regulate the expression of nonmuscle myosin heavy chain (NMMHC) at the levels of transcription and translation. Human surgical specimens including those obtained by endarterectomy and atherectomy will be analyzed by in situ hybridization and immunocytochemistry to localize transcription and translation of nonmuscle myosin isoforms. Northern blots and Dot blots will be used to determine relative abundance of mRNAs. Regulatory elements in the NMMHC-A gene will be identified by transfection experiments and these will be used to characterize nuclear proteins which interact with them. Translational regulation will be explored by isolating cytosolic factors which specifically bind to the large stem-loop structure at the 5 prime end of the NMMHC-A mRNA, and investigating their activity in an in vitro model system. Ultimately, we wish to understand in detail how transcriptional and translational controls together may alter the pattern of nonmuscle myosin expression to produce the proliferative phenotype in smooth muscle.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29AR040580-02
Application #
3457453
Study Section
Cardiovascular and Pulmonary Research A Study Section (CVA)
Project Start
1991-04-01
Project End
1996-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
2
Fiscal Year
1992
Total Cost
Indirect Cost
Name
St. Elizabeth's Medical Center of Boston
Department
Type
DUNS #
073797292
City
Boston
State
MA
Country
United States
Zip Code
01235
Pickering, J G; Jekanowski, J; Weir, L et al. (1994) Liposome-mediated gene transfer into human vascular smooth muscle cells. Circulation 89:13-21
Takeshita, S; Gal, D; Leclerc, G et al. (1994) Increased gene expression after liposome-mediated arterial gene transfer associated with intimal smooth muscle cell proliferation. In vitro and in vivo findings in a rabbit model of vascular injury. J Clin Invest 93:652-61
Losordo, D W; Pickering, J G; Takeshita, S et al. (1994) Use of the rabbit ear artery to serially assess foreign protein secretion after site-specific arterial gene transfer in vivo. Evidence that anatomic identification of successful gene transfer may underestimate the potential magnitude of transgene expressi Circulation 89:785-92
Pickering, J G; Weir, L; Jekanowski, J et al. (1993) Proliferative activity in peripheral and coronary atherosclerotic plaque among patients undergoing percutaneous revascularization. J Clin Invest 91:1469-80
Gal, D; Weir, L; Leclerc, G et al. (1993) Direct myocardial transfection in two animal models. Evaluation of parameters affecting gene expression and percutaneous gene delivery. Lab Invest 68:18-25
Pickering, J G; Bacha, P A; Weir, L et al. (1993) Prevention of smooth muscle cell outgrowth from human atherosclerotic plaque by a recombinant cytotoxin specific for the epidermal growth factor receptor. J Clin Invest 91:724-9
Leclerc, G; Gal, D; Takeshita, S et al. (1992) Percutaneous arterial gene transfer in a rabbit model. Efficiency in normal and balloon-dilated atherosclerotic arteries. J Clin Invest 90:936-44
Nikol, S; Isner, J M; Pickering, J G et al. (1992) Expression of transforming growth factor-beta 1 is increased in human vascular restenosis lesions. J Clin Invest 90:1582-92
Pickering, J G; Weir, L; Rosenfield, K et al. (1992) Smooth muscle cell outgrowth from human atherosclerotic plaque: implications for the assessment of lesion biology. J Am Coll Cardiol 20:1430-9
Leclerc, G; Isner, J M; Kearney, M et al. (1992) Evidence implicating nonmuscle myosin in restenosis. Use of in situ hybridization to analyze human vascular lesions obtained by directional atherectomy. Circulation 85:543-53