Investigating the renal microvasculature in polycystic kidney disease

Lead Research Organisation: University College London
Department Name: Institute of Child Health

Abstract

60,000 people in the UK have long term severe kidney disease. There is currently no cure and patients require dialysis or transplantation. One of the leading causes of long term kidney disease is polycystic kidneys. Our previous work has shown that one of the earliest events in the progressive decline of polycystic kidneys is changes in the small blood vessels of the kidney. Furthermore, we have shown that treatments using proteins which restore and stabilise these blood vessels improve disease progression. However, we do not yet fully understand how these treatments work. This will be investigated in this proposal by a team of scientists and clinicians based at the UCL Great Ormond Street Institute of Child Health and the University of Manchester using high resolution imaging, high-throughput sequencing technology and models which replicate polycystic kidney disease. These investigations will enhance our knowledge of how blood vessels are involved in kidney health and disease. Furthermore, if these studies are successful, it could be a new exciting therapeutic avenue for kidney patients in the future.

Technical Summary

Aims and Objectives: Our aims will be to (i) determine whether gene therapy for VEGFC can treat a autosomal dominant PKD model; (ii) determine the mechanisms underlying VEGFC's effect in PKD.



Methodology: VEGFC will be administered in a model of autosomal dominant PKD by a gene therapy strategy using adeno-associated viruses. We will use a combination of histological and non-invasive imaging to assess the renal microvasculature. We will use transgenic technology to examine whether there are intrinsic defects in kidney endothelia in PKD. In addition, vascular cells lacking PKD genes will be isolated and their biology and gene expression (using RNA-sequencing) assessed. Finally, a 3-dimensional cell culture model will be used to examine if VEGFC directly modulates cystogenesis.



Scientific and Medical Opportunities: This project will enhance our knowledge of renal blood vessels in healthy and polycystic kidneys. In addition, it could lead to the design of new therapies for human renal disease. If this is the case then this will have potential impact on society by improving the quality of life of patients with kidney disease as well as a major economic impact by reducing the number of expensive dialysis regimens or the need for a kidney transplant.

Planned Impact

This research has the potential to discover new treatment strategies for patients with PKD, which accounts for 10% of all cases of end-stage renal disease (ESRD). The UK prevalence of ESRD is 58,000 and every month 600 more people with chronic kidney disease reach ESRD. There is no cure for ESRD patients and they require life-long dialysis and transplantation significantly reducing their quality of life and placing a high burden on healthcare resources with current annual costs of the UK ESRD programme conservatively estimated to be £1 billion or 1% of the total National Health Service budget. Importantly, the therapeutic strategies outlined in this proposal could also be relevant for other renal conditions such as diabetic nephropathy which are also accompanied by changes in the renal microvasculature.

This proposal will take a novel approach and determine whether treatments that target kidney blood vessels can improve PKD. If successful, the work will provide compelling proof of concept evidence that vascular growth factors could be a new treatment strategy to improve this condition. This has the potential to impact on society by improving the quality of life of patients with kidney disease and have an economic impact by reducing the impact of ESRD and saving healthcare costs.

If manipulation of vascular growth factors proves to be a successful treatment for PKD, then this will be of clear interest to pharmaceutical companies. Many companies are interested in vascular growth factor therapy for other conditions and a large amount of compounds to manipulate these signalling pathways are available for therapeutic testing. With specific regard to VEGFC, Lymfactin, an adenovirus developed by Herantis Pharma based on VEGFC gene therapy has been approved in phase 1 clinical trials for breast cancer patients to ameliorate lymphoedema in upper limbs (http://herantis.com/pipeline/lymfactin-for-lymphedema/). Future strategies may involve drug screens to identify alternatie modulators of the microvasculature which could be beneficial in PKD.

Importantly, Dr Long already holds a patent related to targeting the vasculature in PKD and with the help of UCL Business will have regular discussions with UK pharmaceutical companies to licence this intellectual property. If these studies develop new treatments for ESRD then this is likely to bring benefits to the UK economy.

The proposed research will also impact on researchers in multiple scientific disciplines including renal researchers, vascular biologists and immunologists. This is because it should provide insights into: (i) the renal microvasculature in healthy and polycystic kidneys; (ii) the investigation of vascular growth factors as a feasible treatment strategy for PKD (perhaps in combination with vasopressin receptor antagonists) (iii) how adeno-associated virus strategies can be used to treat renal conditions; and (iv) how vascular growth factors alter different cell types and effect downstream signalling pathways. In addition, new tools will be generated which will be of use to renal biologists, vascular scientists and immunologists. Finally, the investigators, post-doctoral researchers and collaborators on the application will directly benefit from the multi-disciplinary nature of the grant involving pre-clinical imaging, molecular and cell biology, gene therapy, transgenic animals and genomics (RNA-sequencing). Collaborators include Professor Mark Lythgoe (pre-clinical imaging), Professor Peter Harris, Professor Paul Beard (photoacuostic imaging) and Dr Simon Waddington (gene therapy).

Publications

10 25 50
 
Description Child Health PhD studentship
Amount £76,334 (GBP)
Organisation Great Ormond Street Hospital (GOSH) 
Sector Hospitals
Country United Kingdom
Start 09/2017 
End 08/2020
 
Description NIHR GOSH Biomedical Centre
Amount £110,000 (GBP)
Funding ID 17BN13 
Organisation Great Ormond Street Hospital (GOSH) 
Department NIHR Great Ormond Street Biomedical Research Centre
Sector Public
Country United Kingdom
Start 04/2017 
End 03/2020
 
Title Cell line - renal collecting duct cells with PKD mutations 
Description Cell line - renal human collecting duct cells with PKD mutations have been generated using CRISPR technology 
Type Of Material Cell line 
Year Produced 2019 
Provided To Others? No  
Impact The cell line is now being used for drug discovery and other therapeutic experiments 
 
Title Epithelial lines with CRIPSR mutation for PKD1 
Description CRISPR mutation for PKD1 generated in HEK293 lines as a cell model for polycystic disease 
Type Of Material Cell line 
Year Produced 2018 
Provided To Others? No  
Impact This cell line is being used to test drugs such as VEGFC which modulate polycystic kidney disease 
 
Title Generation of AAV8 stimulators and inhibitors of VEGFR3 signalling 
Description Generation of AAV8 stimulators and inhibitors of VEGFR3 signalling 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact These reagents allow us to modulate VEGFR3 signalling using an adeno-associated virus with high expression in the kidney 
 
Title Imaging of the developing kidney lymphatics 
Description We have developed tools to image, segment and analyse the vasculature in three-dimensions in developing kidneys 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? No  
Impact Review on this technique in preparation. Used in a study in preparation to E-life. Technique has been provided to other researchers in multiple fields. 
 
Title Imaging of the mircovasculature in cystic kidneys 
Description Imaging of the mircovasculature in cystic kidneys 
Type Of Material Physiological assessment or outcome measure 
Provided To Others? No  
Impact Now we can obtain longitudinal measurements of the renal microvasculature of cystic kidneys 
 
Title Longitudinal measurements of the renal microvasculature using pre-clinical imaging 
Description Longitudinal measurements of the renal microvasculature using pre-clinical imaging 
Type Of Material Data analysis technique 
Provided To Others? No  
Impact The ability to obtain longitudinal measurements of the renal microvasculature using pre-clinical imaging should reduce the number of animals required in these studies. 
 
Description Dr Simon Waddington 
Organisation University College London
Department Institute of Immunity and Transplantation
Country United Kingdom 
Sector Academic/University 
PI Contribution Providing a model to test in-utero gene therapy
Collaborator Contribution Advice on vector design for renal gene therapy. Expertise in in-utero injections of adeno-associated viruses in neonatal mice.
Impact -
Start Year 2015
 
Description Mironid Limited 
Organisation Mironid Ltd
PI Contribution We are testing promising compounds generated by Mironid in cellular and animal models of polycystic kidney disease
Collaborator Contribution Providing compounds to test. Intellectual input into experiments.
Impact None to date
Start Year 2018
 
Description Professor Kari Alitalo 
Organisation University of Helsinki
Country Finland 
Sector Academic/University 
PI Contribution Generation of adeno-associated viruses to stimulate and inhibit the VEGFR3 pathway.
Collaborator Contribution Provision of DNA plasmids
Impact Generation of new adeno-associated viruses
Start Year 2014
 
Description Professor Mark Lythgoe 
Organisation University College London
Department Centre for Advanced Bioimaging
Country United Kingdom 
Sector Academic/University 
PI Contribution Relevant mouse models to use pre-clinical imaging to examine the vasculature in polycystic kidney disease
Collaborator Contribution Imaging tools to examine the vasculature in models of polycystic kidney disease
Impact None yet
Start Year 2015
 
Description Professor Paul Beard 
Organisation University College London
Department Department of Medical Physics and Biomedical Engineering
Country United Kingdom 
Sector Academic/University 
PI Contribution Relevant mouse models to test new computer models to examine the vasculature in polycystic kidney disease
Collaborator Contribution New computer models to examine the vasculature in models of polycystic kidney disease
Impact None yet
Start Year 2016
 
Description Professor Peter Harris 
Organisation Mayo Clinic
Country United States 
Sector Charity/Non Profit 
PI Contribution Analysis of the renal microvasculature in a long-term model of polycystic kidney disease
Collaborator Contribution Provision of transgenic mice
Impact None yet
Start Year 2015
 
Description Vector Biolabs 
Organisation Vector BioLabs
Country United States 
Sector Private 
PI Contribution -
Collaborator Contribution Designing and making adeno-associated viruses for renal gene therapy
Impact -
Start Year 2016
 
Description Careers talk to Kidney Research UK researchers 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Dr David Long gave a talk on career development to a group of young researchers currently funded by Kidney Research UK
Year(s) Of Engagement Activity 2018
 
Description Hosting visit of Kidney Research UK lay advisory board 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Patients, carers and/or patient groups
Results and Impact We organised an event at the Institute of Child Health for the lay advisory committee of Kidney Research UK
Year(s) Of Engagement Activity 2018