Determination of laminin matrix contribution to blood retina barrier function in a novel 3D flow model with application to diabetic retinopathy

The overall aim of this project is to determine the laminin matrix contribution to blood-retinal barrier (BRB) function in a novel three dimensional, tripartite in vitro model of the BRB incorporating fluid flow and diabetic conditions.

The passage of macromolecules between the blood and the inner retinal space is a tightly controlled process that is controlled by a series of structures collectively termed the blood-retina-barrier (BRB). Understanding the basic biology of the BRB is fundamental to understanding the pathogenesis of a number of retinal disorders including diabetic retinopathy, malarial retinopathy and retinitis pigmentosa. Moreover, understanding the regulation of mobility across the BRB is critically required to model the uptake of systemically administered drug into the retina, particularly where normal barrier is compromised.

The BRB is broken into inner and outer parts. In the inner BRB microvascular vessels are encased in a tightly cohesive endothelial tube. These endothelial cells adhere to a basement membrane (BM) on their basal aspect, encased within this BM are retinal pericytes whose signals to the endothelial cells are required to maintain their sheet integrity. The BM is also directly contacted and interacted with by retinal astrocyte foot processes, thereby physically linking the microvasculature to the retinal neural tissue.

Each of the BRB cellular components contributes to the composition and structural organisation of the shared BM. Every BM contains four core component structural networks of laminins and collagens linked by perlecans and nidogens. Of these core components, the greatest structural and functional diversity is within the laminin family. 11 laminin encoding genes giving rise to at least 16 different heterotrimers that are spatially and temporally controlled in their distribution and the precise local combination of which laminins are present defines cell behaviour on that matrix. Each of the cell types involved in the BRB contributes different laminin family members to the shared BM, the importance of which has yet to be defined.

This PhD studentship project will develop an improved in vitro model of the BRB that incorporates all three cellular elements as well as fluid flow to allow in depth molecular dissection of matrix contribution to core cell behaviours in a tractable but still physiologically relevant model.

The specific objectives are;
1) To dissect the contribution of laminin composition and organisation on BRB cell function
2) To determine the influence of fluid dynamics on matrix deposition of BRB cellular components
3) To develop an effective tripartite in vitro model of the BRB that incorporates not only all three cell types but also appropriate matrix and fluid flow conditions.
4) To identify pathological changes of BRB stability in presence of high glucose or due to exposure to AGEs, by studying potential alterations in matrix deposition and BRB cellular behaviour

The student will be supervised by Dr Kevin Hamill, lecturer in molecular biology with a particular focus on laminin biology and Professor Simon Harding, a clinician scientist specialising in retinal disorders.

The Institute of Ageing and Chronic Disease is fully committed to promoting gender equality in all activities. We offer a supportive working environment with flexible family support for all our staff and students and applications for part-time study are encouraged. The Institute holds a silver Athena SWAN award in recognition of on-going commitment to ensuring that the Athena SWAN principles are embedded in its activities and strategic initiatives.

The successful applicant will have (or expect to hold) a First or 2:1 Honours degree in a relevant life science/biomedical subject. A Master’s degree in a relevant area would be an advantage. We are seeking an enthusiastic individual who has a solid understanding of cell and molecular biology..