Identifying therapeutic targets and pathways in diabetic kidney disease

Background.

Kidney disease that results in protein leaking into the urine is a major health issue as it is a prelude to end-stage kidney failure and is also an independent risk factor for cardiovascular complications such as heart attacks and stroke. The leading global cause of this is diabetic kidney disease. This PhD intends to develop in vitro and in vivo transgenic models to understand why this happens and develop new therapies to prevent it.

Outline of project.

Our overall aim is to understand how insulin and insulin-like growth factor I control the behaviour of podocytes and glomerular endothelial cells (cells that form the filtering unit of the kidney glomerulus) in health and disease (specifically the maladaptive changes that underlie insulin resistance). For the in vitro experiments we will explore this in unique conditionally immortalised cell lines (generated at Bristol) and this will be complemented by in vivo work with transgenic mice (notably the Akita mouse model of type-1 diabetes). In addition, in vitro experiments will be undertaken with fluorescent glomerular cells purified from the transgenic mouse tissues by FACS (techniques developed by Prof. Coward- Fig 1). Proteomics will be used to define changes in proteome and phospho-proteome associated with hormone stimulation and resistance (Dr. Heesom is an expert in this field) and functional studies will employ high throughput imaging with a range of fluorescence microscopy reporters (Prof. McArdle is an expert in this field and we have already used this approach to explore insulin signalling in glomerular cells – see reference 1 below).

Translational focus.

It will reveal the critical signalling pathways that are perturbed in the leading cause of kidney failure in the world and identify novel therapeutic targets. It will also lead to the development of new high throughput systems to study glomerular biology, which may also be utilised for a variety of other kidney diseases in the future. Accordingly, the project encompasses both the target identification and the model development that will be needed to further our translational agenda.

How the collaborative arrangements will help to deliver successful PhD training.

The three supervisors in this project are experts in their fields (in vitro and in vivo glomerular biology models - RJC), Development of functional molecular biology cellular systems using the in cell analyser- CAM, and the use and interpretation of state of the art proteomics – KH. Therefore the required support is in place and the project offers an excellent opportunity for a PhD student to be successfully trained in a broad range of important and widely applicable skills.