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.
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/
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