Using imaging and omics approaches to understand the maturing kidney vasculature and enable treatmen

Background:

By birth, the human kidney has developed millions of nephrons that orchestrate renal function. Though human nephron number is set from birth, the kidney grows substantially through childhood, accompanied by changes in the renal microvasculature. The renal microvasculature, comprising specialised blood capillaries and lymphatics, achieves tissue oxygenation, reabsorption, solute and cellular clearance, preventing inflammation (Jafree & Long 2020). Due to congenital mutations, some children suffer from end-stage kidney disease (ESKD). A common pathological feature of which is glomerular damage and leakage of protein into the urine, which does not respond to conventional drug therapies (Tullus et al. 2018). Our group has identified a renal microvascular anomaly in a mouse model of childhood kidney disease. These mice carry a heterozygous mutation in Wilms Tumour 1 (WT1 p.R394W; Gao et al. 2004) found in human patients with congenital glomerular disease. We now want to investigate renal microvasculature maturation and how it changes in childhood kidney disease, in order to identify novel therapeutic approaches to treat ESKD.

Aims/Objectives:

The overall aim of this PhD studentship is to understand how the microvasculature in the kidney develops in childhood and how it is altered in a model of glomerular disease. With this knowledge the student will identify and exploit targeted therapies to support the vasculature and improve kidney function as a novel approach to treat disease.

Objective 1 (0-12 months): Use wholemount immunolabelling for blood vessel and lymphatic markers and cutting-edge 3D imaging to characterise how the renal microvasculature remodels in the postnatal period and how it changes in a Wt1^R394W/+ mouse model of childhood glomerular disease.

Objective 2 (12-24 months): Analyse single-cell RNA sequencing data of Wt1^R394W/+ glomeruli harvested in early disease to identify potential molecular candidates that are altered in the renal microvasculature. Candidate molecules will be tested using iPSC-derived human kidney organoids carrying the same WT1 mutation.

Objective 3 (24-36 months): Follow up molecular candidates screened and selected through Objective 2 and administer these, using gene therapy strategies, to the Wt1^R394W/+ mouse to examine how the therapy affects renal microvasculature and kidney function.

Methods:

The PhD student will have the opportunity to learn techniques ranging from the basics of mouse husbandry, genetics, renal functional assays and histology (Balogh, Chandler, Varga et al. 2020), to newly developed techniques including 3D imaging of lymphatics (Jafree et al. 2019), iPSC and organoid culture, single-cell RNA sequencing analysis and novel therapeutic strategies including gene therapy.

References:

1. Balogh, Chandler, Varga et al. 2020, PNAS, 117: 15137-15147.
2. Gao et al. 2004, Mol Cell Biol., 24: 9899.
3. Jafree et al. 2019, eLife, 8: e48183.
4. Jafree & Long 2020, J Am Soc Nephrol, 31: 1178.
5. Tullus et al. 2018, Lancet Child Adolesc Health, 2: P880.