Project Details
Projekt
Targeting the endothelial glycocalyx to prevent atherosclerosis
Applicant
Dr. Veit Eckardt
Subject Area
Cardiology, Angiology
Term
since 2024
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 547056468
The endothelial glycocalyx as the assembly of glycan carrying cell membrane proteins plays an important role in the development of atherosclerotic plaques at sites of disturbed blood flow. This project attempts to structurally and functionally characterize relevant components of the glycocalyx and explore ways to biochemically stabilize them. Heparan sulfate chains serve as the mechanical levers of laminar blood flow to induce conformational changes in the core proteins which initiate atheroprotective intracellular signaling. By contrast, disturbances in flow result in dysfunctional endothelium which degrades its glycocalyx and becomes permeable to blood cholesterol. A variant in a heparan sulfate-synthesizing enzyme, NDST2, has been associated with myocardial infarction in large GWAS. The glycocalyx may therefore constitute a suitable target for early and site-specific treatment strategies. We plan an in-depth analysis of the endothelial response to different flow patterns in flow chambers by means of RT-PCR, gene expression microarrays and structural analyses of heparan sulfate by gel and high-performance liquid chromatography. To demonstrate an association between glycocalyx structure and endothelial phenotype, we will alter the glycans by enzymatic degradation and knockdown of glycan-synthesizing enzymes and measure changes in endothelial gene expression. These experiments will be performed with endothelial cell lines and induced pluripotent stem cells from donors with the NDST2 risk variant. Moreover, we will create artificially disturbed blood flow by partial ligation of the carotid artery in hyperlipidemic ApoE-/- mice and investigate the endothelial response by single cell RNA sequencing and structural analyses of heparan sulfate. Flow-dependent localized changes will be correlated with atherosclerotic plaques. We then envisage to breed ApoE-/-NDST2-/- double knockout mice and ApoE-/- mice with an endothelium-specific NDST2 knockout and assess their atherosclerotic burden. Finally, we aim to stabilize the glycocalyx for example using covalent attachments of glycan fragments by specific bio-orthogonal reactions. I intend to realize the project at Stanford University because of its scientific focus in both, vascular biology and glycobiology, and its commitment to interdisciplinary translational research. The main host of the project, Prof. Leeper, Chief of Vascular Medicine, has established most of the relevant technologies in his lab. We will closely collaborate with Prof. Bertozzi who was awarded the Nobel Prize in Chemistry in 2022 for her discovery of bio-orthogonal chemistry. After my return to Munich, I aim to pursue a career as clinician-scientist with an independent research group focusing on pathophysiological mechanisms specific for atherosclerosis which may provide the basis for better risk prediction and early treatment strategies.
DFG Programme
WBP Fellowship
International Connection
USA
