KFL Research Grant Report- an update from Richard Naylor

On 14th November 2017 I received funding to the amount of £7500 from the Kidneys for Life charity to purchase a chemical library for screening of drugs in kidney disease models in the zebrafish.

The project was delayed to 2019 as I have been waiting for the zebrafish disease models to come online. I now have disease models for Alport syndrome, a genetic illness caused by mutations in genes encoding for certain forms of Type IV collagen, which is important in proper functionality of the blood filter in the kidney (the glomerulus).

I purchased the chemical library that I budgeted for in the grant application (Stratech FDA-approved chemical library) on the 26th September 2019. The library has now been received and is stored in the deep freeze -80˚C freezer in the Lennon laboratory at the University of Manchester.  The cost of the chemical library had come down to £3357.90 (originally it was £6742 when costing the original application). Thus, I have also used the grant to fund a number of experiments that have enabled me to pump-prime an intermediate fellowship application to Kidney Research UK.

With regards the chemical library screen, in 2020 I will utilise a novel and high throughput readout of glomerular barrier filtration developed in the Lowe laboratory at the University of Manchester. This system will enable me to simply place my disease model fish into the different chemicals of the library and isolate the fish water to test for the presence of a protein (luciferase) that is only present in the fish water if the glomerular filter is defective (due to proteinuria). This readout has the high likelihood of discovering new chemical agents that may be of benefit to the treatment of patients with Alport syndrome.

As mentioned above, the reduction in price of the chemical library offered me the opportunity to perform some preliminary experiments funded by the Kidneys for Life grant money. I have developed a model of polycystic kidney disease and wished to see if kidney tubules isolated from 30 zebrafish embryos was sufficient (in terms of amount of biological material) to perform mass spectrometry analysis for whole proteomics (this experiment cost of £1260). The work proved to be a real success, with over 3100 proteins detected. The ability to be able to do this analysis means that in my fellowship application I am able to convincingly assert that for the first time, I will be able to develop a proteomics signature of polycystic kidney diseased nephron tubules.

The remaining funds from the grant were used to aid more preliminary data analysis (such as purchasing new and more accurate single channel pipettes, an EdU kit to detect cell division (also important in my fellowship application), and purchasing a new Cas9 enzyme to allow me to continue to make my Alport disease models). I also purchased some purified obtustatin, which is a disintegrin peptide that I hypothesise will inhibit integrin α1β1 in the kidney tubule. This inhibition is extremely exciting as it may act to prevent cell division in the tubule and so ameliorate cystogenesis (which is associated with abnormal cell division) and also kidney cancer (renal cell carcinoma).

Taken together, the funding from Kidneys for Life has been immensely helpful in aiding my current projects and also in facilitating preliminary experiments that I hope will drive me towards research independence through the acquisition of an external fellowship. I am sincerely grateful to Kidneys for Life for the awarding of this money to myself and I am keen to continue being a part of the Kidneys for Life community.