(BRC) Defining druggable targets in genetic kidney disease using human pluripotent stem cell -kidney- organoids (Non-Clinical)


   Faculty of Biology, Medicine and Health

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  Prof Sue Kimber, Prof A Woolf  No more applications being accepted  Competition Funded PhD Project (UK Students Only)

About the Project

We and others reported that heterozygous variants of the kidney epithelial transcription factor hepatocyte nuclear factor 1B (HNF1B) are the commonest monogenic causes of human kidney malformations. We created a bank of HNF1B mutant pluripotent stem cell (PSC) lines by CRISPR-Cas9 gene editing wild-type human embryonic stem cells (ESCs) and reprogramming somatic cells to induced PSCs (iPSCs) from HNF1B families followed in Manchester Foundation Trust kidney genetics clinics. Using tailored culture conditions PSCs can be induced to undergo differentiation and form clusters of cells known as organoids which undergo morphogenesis similar to that in human fetal kidney development. We discovered that HNF1B mutant PSC-derived kidney organoids contain malformed and dysfunctional nephron tubules, and we identified glutamate signalling as a key deregulated pathway.

We hypothesise that the aberrant phenotype of HNF1B mutant organoids will be ameliorated by targeting glutamate signalling. We aim to treat HNF1B organoids with commercially available drugs modulating glutamate receptor activity and will assess morphology, gene expression and functionality. The objective is to restore micro-anatomy and functionality. Kidney organoids will be incubated with drugs modulating glutamate receptor activity (e.g., DNQX, a kainate receptor blocker) to determine whether defects in morphology, gene expression and functionality are ameliorated. We will use genetic techniques to knock-down key glutamate receptors in mutant lines, predicting that this will ameliorate their dysmorphology and aberrant biochemistry.

Evaluation will include imaging techniques such as confocal microscopy or light sheet imaging for assessment of morphology and protein expression, transcript evaluation by RT-QPCR and RNAseq together with functional assays such as response to cAMP and calcium imaging. We will also define normal expression patterns of glutamate receptors in developing kidneys from the Wellcome Trust/MRC Human Developmental Biology Resource.

This will define the pathobiology of a rare but clinically important genetic kidney disorder and will be a stepping-stone towards novel treatments for malformed kidneys.

Professor Kimber

https://research.manchester.ac.uk/en/persons/sue.kimber

https://orcid.org/0000-0002-0568-6342

Professor Woolf

https://research.manchester.ac.uk/en/persons/adrian.woolf

https://orcid.org/0000-0001-5541-1358

Eligibility 

Applicants must have obtained or be about to obtain a First or Upper Second class UK honours degree, or the equivalent qualifications gained outside the UK, in a relevant discipline.

Before you Apply 

Applicants must make direct contact with the primary supervisor before applying to discuss their interest in the project. It is your responsibility to make arrangements to meet with potential supervisors, prior to submitting a formal online application.  

How to Apply 

To be considered for this project you MUST submit a formal online application form - full details on how to apply can be found on the BRC website https://www.bmh.manchester.ac.uk/study/research/funded-programmes/manchester-brc-phd-studentships/ 

Your application form must be accompanied by a number of supporting documents by the advertised deadlines. Without all the required documents submitted at the time of application, your application will not be processed and we cannot accept responsibility for late or missed deadlines. Incomplete applications will not be considered. If you have any queries regarding making an application please contact our admissions team.

Equality, Diversity and Inclusion  

Equality, diversity and inclusion is fundamental to the success of The University of Manchester, and is at the heart of all of our activities. The full Equality, diversity and inclusion statement can be found on the website https://www.bmh.manchester.ac.uk/study/research/apply/equality-diversity-inclusion/  

Biological Sciences (4) Medicine (26)

Funding Notes

This studentship covers tuition fees and stipend for home candidates only.

References

Adalat S, Hayes W, Bryant W, Booth J, Woolf AS, Kleta R, Subtil S, Colclough K, Ellard S, Bockenhauer D. HNF1B mutations are associated with a Gitelman-like tubulopathy that develops during childhood. Kidney Int Rep 4:1304-1311, 2019.
Adalat S, Bockenhauer D, Ledermann SE, Hennekam RC, Woolf AS. Renal malformations associated with mutations of developmental genes: messages from the clinic. Pediatr Nephrol 11:2242-2255, 2010.
Adalat S, Woolf AS, Johnstone KA, Wirsing A, Harries LW, Long DA, Hennekam RC, Ledermann SE, Rees L, van’t Hoff W, Marks SD, Trompeter RS, Tullus K, Winyard PJ, Cansick J, Mushtaq I, Dhillon HK, Bingham C, Edghill EL, Shroff R, Stanescu H, Ryffel G, Ellard S, Bockenhauer D. Hepatocyte Nuclear Factor 1B mutations associate with hypomagnesemia and renal magnesium wasting. J Am Soc Nephrol 20:1123-1131, 2009.
Bantounas I, Rooney KM, Lopes FM, Tengku F, Woods S, Zeef LAH, Kuba SY, Bates N, Hummelgaard S, Hillman KA, Silvia Cereghini, Woolf AS, Kimber SJ. Human pluripotent stem cell-derived kidney organoids reveal tubular epithelial pathobiology of heterozygous HNF1B-associated dysplastic kidney malformations. BioRxiv 2023.03.14.532598; doi: https://doi.org/10.1101/2023.03.14.532598