Understanding key molecular signalling pathways in kidney development and disease

Background:
Abnormal development of the kidney and inherited conditions such as Polycystic Kidney Disease (PKD) can lead to end-stage renal failure from birth through adulthood, often requiring dialysis or a kidney transplant. Based on preliminary work done at the University of Toronto and at GOSICH in London, we propose that the ‘Hedgehog’ signalling family is required for normal kidney development while the baby is in the womb and that it is one of the major things disrupted in PKD. Little is currently know about some members of the family, such as Indian hedgehog (IHH), and we postulate that increased understanding might be the first step towards new therapies.

Aims/Objectives:
We will investigate Hedgehog factors using expertise and experimental models available in our two specialised laboratories. Specific objectives are to:
i) Determine the function of hedgehog in nephrogenesis (kidney development), particularly roles in the renal stroma that modulate epithelial structures and nephron number;
ii) Explore roles of diverse hedgehogs in progression of PKD in vivo and in vitro.

Methods:
This project utilises multiple techniques from animal husbandry through morphological and immunohistological analysis to standard and innovative molecular assays such as Western blotting, Real-time PCR, RNA-scope and dropseq. We will also generate novel PKD cell lines using CRISPR-Cas9 technology for 3-dimensional culture to examine the effects of drugs or molecular manipulation of hedgehogs (siRNA, overexpression) on cyst numbers and size.

Collaboration with University of Toronto:
Work in Toronto is an intrinsic part of this project. The student will be attached to Professor Rosenblum’s research group for 6-9 months in year 2. Professor Rosenblum has extensive experience in both human and mouse studies and has all of the transgenic strains and resources required to breed complex mutant mice to assess the roles of hedgehogs in development, initially via morphology and histology but also by learning novel dropseq techniques already set up Toronto. Cell lines will also be generated to bring back to the UK for in vitro experiments to complete the PhD.

Timeline:
First and third year will be based at GOSICH with 6-9 months spent in Toronto in year 2. Exact timing for the Canadian part will depend on when the transgenic strains are ready for analysis - these will be generated by Prof Rosenblum’s team ahead of the placement.

References:
1. Huang JL et al J Am Soc Nephrol 27: 69-77, 2016.
2. Brzoska et al Kidney Int 90: 1274-1284, 2016.
3. Price KL et al Cell Death and Discovery 4:13. doi: 10.1038/s41420-017-0021-6, 2018
4. Rowan CJ, et al Development. 145(13). pii: dev159947. doi: 10.1242/dev.159947, 2018.
5. Sheybani-Deloui S et al J Am Soc Nephrol. 29: 532-544, 2018.