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.
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 alternative 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).
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 alternative 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).
Organisations
- University College London (Lead Research Organisation)
- University College London (Collaboration)
- Mayo Clinic (Collaboration)
- Vector BioLabs (Collaboration)
- Washington University in St. Louis (Collaboration)
- University of Helsinki (Collaboration)
- Mironid Ltd (Collaboration)
- University of Toronto (Collaboration)
Publications
Ogunlade O
(2018)
In vivo three-dimensional photoacoustic imaging of the renal vasculature in preclinical rodent models.
in American journal of physiology. Renal physiology
Jafree DJ
(2021)
Mechanisms and cell lineages in lymphatic vascular development.
in Angiogenesis
Price KL
(2018)
Lithium induces mesenchymal-epithelial differentiation during human kidney development by activation of the Wnt signalling system.
in Cell death discovery
Perretta-Tejedor N
(2020)
Endothelial-epithelial communication in polycystic kidney disease: Role of vascular endothelial growth factor signalling.
in Cellular signalling
Henderson DJ
(2018)
Planar cell polarity in organ formation.
in Current opinion in cell biology
Hernandez-Diaz I
(2019)
Overexpression of Circulating Soluble Nogo-B Improves Diabetic Kidney Disease by Protecting the Vasculature.
in Diabetes
Morais MRPT
(2022)
Kidney organoids recapitulate human basement membrane assembly in health and disease.
in eLife
Vasilopoulou E
(2018)
Thymosin-ß4: A key modifier of renal disease.
in Expert opinion on biological therapy
Papakrivopoulou E
(2021)
The Biological Significance and Implications of Planar Cell Polarity for Nephrology.
in Frontiers in physiology
Lim VG
(2019)
SGLT2 Inhibitor, Canagliflozin, Attenuates Myocardial Infarction in the Diabetic and Nondiabetic Heart.
in JACC. Basic to translational science
Bryant D
(2022)
The timing of auditory sensory deficits in Norrie disease has implications for therapeutic intervention.
in JCI insight
Randles MJ
(2021)
Identification of an Altered Matrix Signature in Kidney Aging and Disease.
in Journal of the American Society of Nephrology : JASN
Jafree DJ
(2020)
Beyond a Passive Conduit: Implications of Lymphatic Biology for Kidney Diseases.
in Journal of the American Society of Nephrology : JASN
Sanchis P
(2019)
Arterial "inflammaging" drives vascular calcification in children on dialysis.
in Kidney international
Perretta-Tejedor N
(2020)
Generating Mutant Renal Cell Lines Using CRISPR Technologies.
in Methods in molecular biology (Clifton, N.J.)
Jafree DJ
(2020)
Tissue Clearing and Deep Imaging of the Kidney Using Confocal and Two-Photon Microscopy.
in Methods in molecular biology (Clifton, N.J.)
Walsh CL
(2021)
Imaging intact human organs with local resolution of cellular structures using hierarchical phase-contrast tomography.
in Nature methods
Mason WJ
(2022)
Systemic gene therapy with thymosin ß4 alleviates glomerular injury in mice.
in Scientific reports
Papakrivopoulou E
(2018)
Vangl2, a planar cell polarity molecule, is implicated in irreversible and reversible kidney glomerular injury.
in The Journal of pathology
Description | Child Health PhD studentship |
Amount | £76,334 (GBP) |
Organisation | Great Ormond Street Hospital (GOSH) |
Sector | Hospitals |
Country | United Kingdom |
Start | 08/2017 |
End | 08/2020 |
Description | Dissecting the role of macrophages in kidney development |
Amount | £153,536 (GBP) |
Funding ID | Paed_RP_010_20180928 |
Organisation | Kidney Research UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 08/2019 |
End | 09/2021 |
Description | Exploring the contribution of lymphatics towards diabetic kidney disease and their potential as a therapeutic target |
Amount | £107,294 (GBP) |
Organisation | Diabetes UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 03/2020 |
End | 03/2023 |
Description | Lymphatic biology in kidney development, health and disease |
Amount | £1,555,838 (GBP) |
Funding ID | 220895/Z/20/Z |
Organisation | Wellcome Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 08/2020 |
End | 08/2025 |
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 | Academic/University |
Country | United Kingdom |
Start | 03/2017 |
End | 03/2020 |
Description | Single-cell transcriptomics of the renal vasculature in polycystic kidney disease |
Amount | £14,960 (GBP) |
Organisation | PKD Charity |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 06/2022 |
End | 12/2023 |
Description | Spatial mapping and single-cell transcriptomics of kidney lymphatics in WIlm's tumour |
Amount | £48,697 (GBP) |
Funding ID | PGS19-2/10174 |
Organisation | Rosetrees Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 03/2020 |
End | 03/2022 |
Description | Using nanobodies for labelling and threedimensional imaging of intact adult mouse kidneys at single-cell resolution |
Amount | £36,142 (GBP) |
Organisation | Kidney Research UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 08/2019 |
End | 08/2021 |
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 | Benjamin Humphreys |
Organisation | Washington University in St Louis |
Country | United States |
Sector | Academic/University |
PI Contribution | We have analysed this dataset and identified a unique population of vascular cells with unique molecular features within human PKD kidneys. |
Collaborator Contribution | Professor Humphreys and his laboratory have generated a single-cell RNA-sequencing dataset of human PKD (https://www.nature.com/articles/s41467-022-34255-z). This dataset involves reading the cell-by-cell gene expression of 17,000 cells, enabling us to assess the molecular features and 'behaviour' of blood vessels in humans with PKD. |
Impact | Enhancing our research on the kidney vasculature |
Start Year | 2022 |
Description | Christiana Ruhrberg |
Organisation | University College London |
Department | Institute of Ophthalmology UCL |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Intellectual input and imaging technology for evaluating lymphatics in developing kidneys. |
Collaborator Contribution | Intellectual input and mouse lines to explore the cellular origins of kidney lymphatics |
Impact | Publication in e-life in 2019 |
Start Year | 2019 |
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 |
Country | United Kingdom |
Sector | Private |
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 | Norman Rosenblum |
Organisation | University of Toronto |
Country | Canada |
Sector | Academic/University |
PI Contribution | Intellectual input and techniques to examine lymphatics in developing kidneys |
Collaborator Contribution | Intellectual input and transgenic mice to examine the cellular origins of kidney lymphatics |
Impact | Visit of PhD student to Professor Rosenblum's laboratory New PhD student appointed between UCL and Toronto laboratory |
Start Year | 2019 |
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 | Simon Walker-Samuel collaboration |
Organisation | University College London |
Department | Centre for Advanced Bioimaging |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Intellectual input and imaging data to establish mathematical models of lymphatic development in the kidney |
Collaborator Contribution | Intellectual input and the establishment of mathematical models of lymphatic development in the kidney |
Impact | Publication of manuscript in eLife. |
Start Year | 2018 |
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 | GOSICH BRC Family Fun Day |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Our group ran an interactive activity at this event called "How the kidney works?" together with Kidney Research UK. It was well attended and we received excellent feedback from the participants. |
Year(s) Of Engagement Activity | 2019 |
URL | https://www.gosh.nhs.uk/research-and-innovation/nihr-gosh-brc/brc-events/nihr-great-ormond-street-ho... |
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 |