Vascular mechanotransduction has long been recognized as mediating physiological processes such as flow-dependent dilation and pathological processes such as the localization of lesions in atherosclerosis. There is a consensus that hemodynamic forces represent two distinct biomechanical stimuli;that is, while unidirectional laminar fluid shear is atheroprotective, rapidly changing and/or reverse fluid shear stress (coupled with low average shear) is atherogenic. The overall goal of this application is to determine the molecular basis of mechanochemical signal transduction and the endothelium's ability to discriminate between these two biomechanical stimuli. We have already demonstrated that the atherogenic/inflammatory hemodynamic signaling originates from a macromolecular complex assembled around PECAM-1 at the endothelial cell-cell junction. One of the objectives of this application is to define the organization of the macromolecular complex located at the endothelial cell-cell junction and elucidate its mechanism of activation. The second objective is to investigate the hypothesis that the structural geometry of the junction regulates the sensitivity of the macromolecular complex to rapidly changing shear and confers the ability to discriminate between unidirectional and oscillatory/reverse flow. This project will use an entirely integrative approach using molecular biology, cell biology and biochemistry, vital cell imaging, and cell biomechanics. In particular, the specific aims are: 1) To investigate the interaction of PECAM-1 with VE-cadherin and VEGFR2 at the molecular level and the regulation of mechanochemical signaling through this interaction;2) Determine the role of junctional GPCRs and G-proteins in mechanochemical signaling and their regulation by PECAM-1;3) Characterize the geometry of the cell-cell junction by measuring its angle of inclination in in vivo and flow-adapted in vitro endothelium;and 4) Determine whether junctional inclination regulates endothelial response to reverse and oscillatory flow. The understanding of the molecular and structural mechanisms by which the endothelium senses oscillatory and reverse flow is crucial for the development of targeted therapies for flow-induced vascular inflammation and atherogenesis.

Public Health Relevance

Atherosclerosis occurs at regions of the vasculature where blood flow is highly unsteady and oscillatory, suggesting that this blood flow pattern promotes the disease. This project will determine how this blood flow pattern induces the signals that lead to atherosclerosis. If successful, it will provide a basis for drug design for its treatment.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
3R37HL040696-28S1
Application #
8920888
Study Section
Bioengineering, Technology and Surgical Sciences Study Section (BTSS)
Program Officer
Fleg, Jerome
Project Start
1988-04-01
Project End
2019-06-30
Budget Start
2014-09-05
Budget End
2015-06-30
Support Year
28
Fiscal Year
2014
Total Cost
$164,704
Indirect Cost
$73,354
Name
La Jolla Institute
Department
Type
DUNS #
114215473
City
La Jolla
State
CA
Country
United States
Zip Code
92037
dela Paz, Nathaniel G; Melchior, Benot; Shayo, Francisca Y et al. (2014) Heparan sulfates mediate the interaction between platelet endothelial cell adhesion molecule-1 (PECAM-1) and the G?q/11 subunits of heterotrimeric G proteins. J Biol Chem 289:7413-24
Melchior, Benoit; Frangos, John A (2014) Distinctive subcellular Akt-1 responses to shear stress in endothelial cells. J Cell Biochem 115:121-9
Kalyanaraman, Hema; Schwappacher, Raphaela; Joshua, Jisha et al. (2014) Nongenomic thyroid hormone signaling occurs through a plasma membrane-localized receptor. Sci Signal 7:ra48
dela Paz, Nathaniel G; Melchior, Benoit; Frangos, John A (2013) Early VEGFR2 activation in response to flow is VEGF-dependent and mediated by MMP activity. Biochem Biophys Res Commun 434:641-6
Orjuela-Sanchez, Pamela; Ong, Peng Kai; Zanini, Graziela M et al. (2013) Transdermal glyceryl trinitrate as an effective adjunctive treatment with artemether for late-stage experimental cerebral malaria. Antimicrob Agents Chemother 57:5462-71
Ong, Peng Kai; Melchior, Benoit; Martins, Yuri C et al. (2013) Nitric oxide synthase dysfunction contributes to impaired cerebroarteriolar reactivity in experimental cerebral malaria. PLoS Pathog 9:e1003444
Melchior, Benoit; Frangos, John A (2010) Shear-induced endothelial cell-cell junction inclination. Am J Physiol Cell Physiol 299:C621-9