Project 3

Background & Objective

Large artery stiffening leads to systolic hypertension and is accelerated in diabetes. Advanced glycation endproducts (AGEs) are implicated in normative ageing, diabetes, and hypertension. Methylglyoxal (MGO) is a major precursor of AGE formation and MGO can be detoxified by glyoxalase-1 (Glo1). At the population level, plasma MGO is associated with increased systolic hypertension. The various pathways through which MGO could exert its effect on (accelerated) arterial stiffening have yet to be established. Possible routes include extracellular matrix cross-linking, vascular cell phenotypic changes, and perivascular tissue modulation of smooth muscle tone. This project is aimed at establishing how MGO modulates the cellular ageing process and, thereby, the arterial stiffening process.

Methodology

• Experimental models of Glo-1 overexpression and of ageing (smooth muscle and endothelial cell specific Ercc1∆/- mice) will be used.
• Biological and physiological effects of MGO across molecular-cell-tissue-vessel scales will be studied. Vascular reactivity will be determined by advanced pressure myography. Specific markers of cellular senescence will be included. 
• We have developed in-house a dedicated setup to quantify arterial biaxial mechanical properties as well as microscopic ultrastructure. In the project, a new approach needs to be developed to quantify smooth muscle tone ex vivo in intact vessel segments. In addition, smooth muscle cells will be characterised in vitro, enabling detailed phenotyping, including myogenic response and contractile markers like SM-MHC and smoothelin.
• We will include methods to capture and identify the paracrine role of perivascular fat tissue. MGO has been shown to impact perivascular fat metabolism and, in turn, perivascular fat has been shown to modulate smooth muscle cell function and phenotype.
• In addition to mechanistic studies, a pyridoxamine (vitamin B6)-based therapeutic approach to quench MGO will be developed and tested.

Collaborative secondments
For the project, three supporting secondments are planned to develop skills and widen the scientific scope. These include 2 months at the Fernandez-Alfonso lab at UCM on perivascular adipose tissue assays, 1 month at the Shiels lab at UGLA on cellular senescence assays, and 1 month at Quipu Srl on vascular ultrasound image quantification, in collaboration with MINDSHIFT projects 11 and 7.

Requirements

• Qualifications: Master in biochemistry, experimental biology, or equivalent
• Experience: Preferably combinations of: biological experimentation - tissue preparation - pharmacological studies - operating complex analytical equipment
• Knowledge & skills:Cell culture, physiological measurements; in combination with animal research certification would be helpful
• Abilities: Good communication skills, project planning and time management, integration of multi-disciplinary information and context
• Attitude and disposition: Highly enthusiastic about research and willing to learn; creative and independent attitude; team worker