Autosomal dominant polycystic kidney disease (ADPKD) is the most frequent Mendelian inherited kidney disease (prevalence 1/1000). The disease is characterised by hypertension, increased cyst formation, enlarged kidney size, pain and progressive decline to end stage renal failure. Genetic mutations in PKD1 or PKD2 are responsible for disease in most patients.
PKD1 and 2 code for polycystin 1 and 2 respectively. These proteins have important functions in primary cilia in kidney tubules. We propose using nanoparticle technology to selectively target these genes to the kidneys. Nanoparticles are a non-viral method of delivering genes for gene therapy. They have a large packaging capacity, are non-immunogenic, have low toxicity and are low risk for insertional mutagenesis. We have a collaboration in place with 4BaseBio, who have expertise in nanoparticles that will facilitate progression of this project.
A recent publication using a conditional knockout PKD2 mouse model, reintroduced the PKD2 gene when mice had already developed cysts, and demonstrated that there is a capacity for repair and regeneration in the adult mouse kidney (Dong et al 2021). This supports the concept that gene therapy introduced after disease onset, is a feasible option to treat ADPKD.
Aims and Objectives
The key hypothesis is:
- Nanoparticle gene therapy can effectively and specifically transduce kidney tubular cells to slow the disease phenotype in autosomal dominant polycystic kidney disease.
The project’s primary objectives will be to
- show nanoparticle transduction of kidney tubular cells in vitro
- develop tubular cell specific nanoparticles in conjunction with our collaborators, 4basebio.
- test nanoparticle transduction in vivo in wild type mice
- test nanoparticles expressing wild type PKD2 in human cells with a PKD2 mutation, and in mice with a conditional knockout of PKD2.
The student will develop skills in planning, and carrying out both in vitro and in vivo experiments.
The student will learn cell culture techniques and how to use nanoparticles to transduce kidney cells. To determine efficiency of nanoparticle transduction, we will use flow cytometry, western blotting, and immunofluorescence. There will be opportunities to learn these techniques as well as to learn how to use wide field and confocal microscopy. There might be opportunities to visit 4basebio and be involved in the development of nanoparticles for tubular kidney cells.
There will be opportunities to learn about in vivo work, including learning about how to genotype and manage genetic mouse model colonies. We will be testing how well the nanoparticles work in vivo using intravenous injections. Animal tissue will then be analysed at the end of experiment, using quantitative PCR, RNAScope, western blotting, immunofluorescence. This will provide the student with a broad range of laboratory skills.
Nanoparticle, Gene Therapy, Kidney Disease
How to apply for this project
This project will be based in Bristol Medical School - Translational Health Sciences in the Faculty of Health Sciences at the University of Bristol.
Please visit the Faculty of Health Sciences website for details of how to apply