Pomeranz, Gideon; (2024) Experimental models to understand diabetic vascular complications. Doctoral thesis (Ph.D), UCL (University College London).

Text Experimental models to understand diabetic vascular complications.pdf - Accepted Version Access restricted to UCL open access staff until 1 June 2027. Download (53MB)

Abstract

Diabetes mellitus (DM) is a metabolic disorder which leads to vascular complications including retinopathy, nephropathy, and neuropathy through exposure to continued hyperglycaemia. To understand how these complications occur and develop new treatments, reliable experimental models are required to study the effects of excessive glucose on blood vessels. I hypothesised that 3-dimensional blood vessel organoids (BVOs) could be used to model diabetic vascular complications. Firstly, I independently replicated a published protocol to generate BVOs in my host laboratory. Using microinjection techniques, I was able to show that these organoids have liquid carry and transport capabilities. To model diabetes, BVOs were then exposed to either 25mM exogenous glucose or 7.5% serum from diabetic patients with or without nephropathy. Changes in vascular networks were assessed using a quantitative image analysis pipeline, to measure the mean radius and length of vessels within the BVO, as well as overall statistics of the network such as volume, network length and number of segments. Using this pipeline, excessive glucose caused a significant decline in vessel radii while the inverse was true for BVOs treated with human serum which was accompanied by altered TGFbeta signalling Next, to find potential molecules involved in vascular complications, I analysed multiple single cell RNA sequencing datasets to assess changes in endothelial gene expression in BVOs treated with excessive glucose as well as mice and humans with diabetic nephropathy. In parallel, I generated a transgenic zebrafish expressing vitamin D binding protein tagged to GFP, which when treated with 25mM of glucose resulted in vascular leakage with lower dorsal aorta fluorescence, establishing an in- vivo model to study diabetic vascular complications. Collectively, this thesis presents two experimental approaches to study diabetic vascular complications and highlights how computational techniques which interrogate multiple datasets can identify candidate molecules for therapeutic testing in this context.

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