Migration of vascular smooth muscle (VSM) cells occurs during development, wound healing, angiogenesis, and contributes to the progression of vascular disease. Published studies support a role for the ubiquitous multifunctional serine/threonine protein kinase, Ca2+/calmodulin-dependent protein kinase II (CaMKII in regulating VSM cell migration, but the mechanisms of CaMKII activation in this setting, and the relevant substrate targets and mechanisms underlying effects on migration, are unknown. Based on our published and preliminary studies we have hypothesized that localized leading edge activation of CaMKII42 regulates focal adhesion dynamics and acts as a positive feedback to promote VSM cell polarization and directional migration.
In Aim 1 molecular/genetic approaches will be used to manipulate CaMKII42 expression and activity, and the consequences on directional persistence and velocity of individual migrating VSM cells determined. Fluorescence recovery after photobleaching (FRAP) analysis of GFP-tagged focal adhesion components will be used to assess CaMKII4-dependent regulation of focal adhesion dynamics.
In Aim 2 biochemical and advanced fluorescence imaging approaches (confocal FRET and TIRF microscopy) will be used to test a potential mechanism involving a functional CaMKII42 interaction with the Src-family kinase, Fyn, in regulating focal adhesion protein dynamics. VSM cell migration is a property of """"""""synthetic phenotype"""""""" cells that have not acquired, or have lost, differentiated contractile protein markers and function. We have determined that up-regulation of a CaMKII4 isoform (42 or 4C) contributes to VSM cell synthetic phenotype functions (proliferation, migration). Recent studies indicate up-regulation of certain Ca2+ conducting cation channels in the TRPC and STIM/Orai families in synthetic phenotype VSM cells is linked functionally to regulation of proliferation and migration. Based on this, in Aim 3 we molecular and fluorescence imaging approaches to test the hypothesis that STIM1/Orai1 mediated Ca2+ entry is functionally linked to leading edge CaMKII activation.
In aim 4 we test the hypothesis that that expression of CaMKII4 isozymes and these Ca2+ channels are co-regulated as a function of VSM phenotype, conferring an integrated pathway for Ca2+-dependent regulation of VSM cell migration in vivo. Results of these novel studies are expected to provide insight into mechanisms by which Ca2+ signals regulate VSM cell migration and to provide a basis for understanding Ca2+-dependent regulation of migration in other cell types and processes such as myofibroblast-mediated wound healing or angiogenesis. Taken with recently published studies in heart and skeletal muscle, these studies in VSM suggest that CaMKII4 isoforms may be generally important in muscle remodeling in response to physiological or pathophysiological stressors.

Public Health Relevance

By elucidating the functional consequences of CaMKII isoforms in response to vascular injury, these studies are expected to provide insights into mechanisms underlying vascular proliferative diseases including atherosclerosis and restenosis follow vascular surgery. Because Ca2+ signals and CaMKII are a ubiquitous regulatory system in all cells and tissues, the information derived from these studies can be reasonably expected to extend to other processes involving vascular cell motility such as myofibroblast-mediated wound healing or angiogenesis.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL092510-01A1
Application #
7899534
Study Section
Vascular Cell and Molecular Biology Study Section (VCMB)
Program Officer
Gao, Yunling
Project Start
2010-03-15
Project End
2014-02-28
Budget Start
2010-03-15
Budget End
2011-02-28
Support Year
1
Fiscal Year
2010
Total Cost
$395,000
Indirect Cost
Name
Albany Medical College
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
190592162
City
Albany
State
NY
Country
United States
Zip Code
12208
Lu, Yao Wei; Lowery, Anthony M; Sun, Li-Yan et al. (2017) Endothelial Myocyte Enhancer Factor 2c Inhibits Migration of Smooth Muscle Cells Through Fenestrations in the Internal Elastic Lamina. Arterioscler Thromb Vasc Biol 37:1380-1390
Li, Jingjing; Miao, Lianjie; Shieh, David et al. (2016) Single-Cell Lineage Tracing Reveals that Oriented Cell Division Contributes to Trabecular Morphogenesis and Regional Specification. Cell Rep 15:158-170
Saddouk, Fatima Z; Sun, Li-Yan; Liu, Yong Feng et al. (2016) Ca2+/calmodulin-dependent protein kinase II-? (CaMKII?) negatively regulates vascular smooth muscle cell proliferation and vascular remodeling. FASEB J 30:1051-64
Liu, Yong Feng; Spinelli, Amy; Sun, Li-Yan et al. (2016) MicroRNA-30 inhibits neointimal hyperplasia by targeting Ca(2+)/calmodulin-dependent protein kinase II? (CaMKII?). Sci Rep 6:26166
Spinelli, Amy M; Liu, Yongfeng; Sun, Li-Yan et al. (2015) Smooth muscle CaMKII? promotes allergen-induced airway hyperresponsiveness and inflammation. Pflugers Arch 467:2541-54
Simone, Tessa M; Higgins, Stephen P; Archambeault, Jaclyn et al. (2015) A small molecule PAI-1 functional inhibitor attenuates neointimal hyperplasia and vascular smooth muscle cell survival by promoting PAI-1 cleavage. Cell Signal 27:923-33
Zhao, Chen; Guo, Hua; Li, Jingjing et al. (2014) Numb family proteins are essential for cardiac morphogenesis and progenitor differentiation. Development 141:281-95
Cleary, Rachel A; Wang, Ruping; Waqar, Omar et al. (2014) Role of c-Abl tyrosine kinase in smooth muscle cell migration. Am J Physiol Cell Physiol 306:C753-61
Ginnan, Roman; Jourd'heuil, Frances L; Guikema, Benjamin et al. (2013) NADPH oxidase 4 is required for interleukin-1?-mediated activation of protein kinase C? and downstream activation of c-jun N-terminal kinase signaling in smooth muscle. Free Radic Biol Med 54:125-34
González-Cobos, José C; Zhang, Xuexin; Zhang, Wei et al. (2013) Store-independent Orai1/3 channels activated by intracrine leukotriene C4: role in neointimal hyperplasia. Circ Res 112:1013-25

Showing the most recent 10 out of 20 publications