Dr Maria Kaisar – Developing new ways to assess kidneys so transplants last for longer.

With funding from Kidney Research UK, a team of researchers from the University of Oxford, the University of Nottingham and University College London will develop ways to assess donor kidneys and predict how well they will work after transplant.

Having a kidney transplant is the best treatment for kidney failure, but the demand for donated kidneys is high.

To save more lives, doctors are now accepting kidneys from older or higher risk donors. These kidneys may also work less well after transplantation. This can have devastating effects, causing patients once recovering from transplantation to also go back on to dialysis, and wait for another transplant.

Right now, doctors cannot accurately assess donor kidneys. This makes it difficult to predict how well a transplant will work and how long a kidney will last after it has been transplanted.

Thanks to Kidney Research UK’s grant award of £237,626, (in partnership with the Stoneygate Trust), the ADMIRE study ‘Assessing Donor kidneys and Monitoring Transplant Recipients’ aims to address this clinical challenge.

Dr Maria Kaisar

Dr Maria Kaisar from Oxford University’s Nuffield Department of Surgical Sciences (NDS) is the Principal Investigator on the study and leads a team of co-investigators from NDS (Professor Rutger Ploeg, Dr Edward Sharples, Mr Simon KnightMr James Hunter and Dr Sadr Shaheed), the Oxford Big Data Institute (Dr Alberto Santos Delgado and Dr Philip Charles) and the Radcliffe Department of Medicine (Dr Elizabeth Tunnicliffe)

Dr Maria Kaisar and her team will utilise the Oxford Transplant Biobank (OTB) and the Quality in Organ Donation (QUOD) biobank to look for marker proteins in the donors’ blood samples. They use these samples to develop a mathematical model to predict how well donor kidneys will work after transplantation. The successful model would allow doctors to accurately assess kidneys and only transplant those that will function well. It could also identify suitable kidneys previously deemed too high risk to transplant.

With Professors Sue Francis and David Long from the University of Nottingham and University College London, the NDS team will use the QUOD X platform to also develop a monitoring strategy. MRI scanning methods will be performed on both the donor organ before it is transplanted, and later on after transplantation. This will allow us to monitor how well the transplanted organ is functioning.

“I am absolutely delighted that our study received this funding award by Kidney Research UK in partnership with The Stoneygate Trust,” said Maria. “This funding will enable us to bring scientific and clinical expertise together in collaboration, to develop novel non-invasive methods to better assess donor kidneys and, predict how well a transplant will work in the recipient. We also envisage that our planned scientific work will offer many opportunities to our early career scientists, to further develop their skills and research expertise in studying kidney disease. “

Letizia Lo Faro – Use of QUOD samples in research ‘Case Study’

My name is Letizia Lo Faro and I am a Post-Doctoral research scientist in the Oxford Transplant research group. I have so far used QUOD samples in 4 or 5 separate research studies. In one of these, conducted together with Ms Flavia Neri (University of Padova), we were interested in studying the molecular features of donor kidneys with acute kidney injury (AKI) having different functional outcomes after transplant.

Letizia Lo Faro

Once we fully characterised our research question, we moved onto donor sample selection. We decided to compare 4 different groups of donors: with or without AKI and each with good (≥45 mL/min) or poor (<45 mL/min) function (eGFR) 12 months post-transplant, for a total of 40 donors. We maximised the difference between outcomes as much as possible and to allow for fair comparisons we decided to match the groups for several other variables (donor age and gender, BMI, cold ischaemia time, recipient age, recipient gender…just to name a few). I believe appropriate sample selection is a key step in such retrospective studies and we engaged with the QUOD Data Manager earlier on in the process. This provided us with a great overview of all the variables available and also made sure our groups were nicely balanced.

Once we were happy with the donor selection, we proceeded with requesting the samples from the biobank. In this case they were full RNAlater frozen kidney biopsies and formalin-fixed paraffin-embedded (FFPE) kidney tissue slides. In this study we were interested in studying protein expression, so first of all we processed the tissue biopsies to mechanically homogenise them and extract the proteins. Later on the proteins were quantified and the expression of 17 proteins of interest was analysed by Western Blotting (a technique where a mixture of proteins is separated on a gel, based on the molecular weight, and, following transfer on a membrane, proteins of interest are identified by binding to specific antibodies). Results were then analysed with statistical tools.

A set of FFPE slides was then utilised for histological assessment, by standard staining with haematoxylin and eosin (H&E), to quantify chronic and acute tissue damage. One additional set of slides was utilised for confirmation of the western blotting results by immunohistochemistry, another method which allows to check for the presence of certain proteins/products in a tissue, by binding with labelled antibodies.

Results from this study were presented at the British Transplantation Society meeting last year and we are currently finalising a manuscript for publication.

Our results suggested that specific molecular patterns are recognizable in acutely inured kidneys that proceed towards worse function after transplantation. In particular, Peroxisome proliferator-activated receptor gamma (PPAR gamma) specifically increased after acute injury that progressed to worse function, underlining a potential role of metabolic dysfunction in the development of kidney disease (Figure 1).

Figure 1. The protein PPARg, quantified by western blotting analysis, is significantly increased in AKI kidneys with poor outcomes 12months post-transplant.

Our findings have helped identify potential molecular mechanisms involved in the progression of acute kidney injury to chronic kidney disease and post-transplant dysfunction and may constitute a therapeutic target of further interventions aimed at improving the quality of donated kidneys with an acute injury.

Working with QUOD samples has allowed us to really dive deep into the study of molecular mechanisms of kidney injury and the availability of different sample types also allowed us to apply various techniques and methods to validate our findings.

New QUOD Colleague Questionnaire: Philippa Wren – QUOD & Transplant Research Groups Administrative Assistant

In June, QUOD welcomed our new QUOD & Transplant Research Groups Administrative Assistant – Philippa Wren. Philippa has kindly agreed to share a bit more about herself and her role at QUOD:

Philippa Wren

What were you doing most recently before joining QUOD?
Surviving lockdown and home schooling my beautiful daughters, Grace 9, and Evelyn 2. My previous role involved PA, Administrative and Secretarial support for a Regional Manager. I also have a background in Marketing and Events organising.

What interested you about working with QUOD?
I have missed working within a team and hoped to work within a reputable academic organisation. I have always been fascinated with surgery and liked the thought of working somewhere that makes a difference. The two together seemed like the perfect opportunity for me..

What does your role in QUOD involve?
I provide administrative support for the QUOD and Transplant Research teams. I maintain the contact@quod inbox, social media platforms and website as well as take minutes for various meetings. I also place orders, liaise with suppliers of lab consumables and support Prof Ploeg’s EA in looking after his diary, organising his meetings and travel arrangements.

Lightning round time. Tea or coffee?
Tea – I love it!

Favourite place in Oxford?
There are so many. I would have to say the view of Oxford Skyline from the top of South Parks.

Best holiday trip ever?
Venice. There’s no other place quite like it.

Primary COVID-19 lockdown survival method?
Lots of long walks… and wine!

Finally, what kinds of inquiries should people bring to you, and how best can they reach you?
All inquiries are welcome. If I can’t answer it I can redirect you to someone who can. You can contact me via email contact@quod.org.uk or philippa.wren@nds.ox.ac.uk

Spotlight on a QUOD Colleague – Maggie Stevens

by Susan Patchett

For this edition of the QUOD Newsletter I was delighted to be able to catch up with Maggie Stevens, ODT Specialist Nurse for Research at NHS Blood and Transplant.  Maggie is responsible for the operational and service delivery for the great number of research studies that NHS Blood and Transplant operate and support alongside QUOD.

Hers is a varied role and involves setting up Specialist Nurse training, carrying out risk assessments for new studies, and helping teams draw up action plans and standard operating procedures (SOPS) ensuring that they comply with their ethics approval, regulatory requirements, and are fit for purpose for Specialist Nurses in Organ Donation (SNODs) to follow safely and effectively.

When I asked Maggie if she could describe a typical day she told me that there was no such thing.  One day she can be going through a risk assessment with the ODT Research Team and Quality Assurance for a large clinical trial and identifying potential operational, regulatory and safety risks.  Other days she will be auditing consent records to be sure all has been carried out to the letter.  She meets regularly with research leads in all the regions, ensuring they are up to date with studies that are being set up and also cascading training that they will undertake with their teams. 

Problem solving is a large part of her role and can range from SNODs raising issues if they aren’t comfortable with a particular process, to a shortage of boxes. 

Maggie works very closely with QUOD and helps us iron out issues that may arise in terms of sample collection, logistics, and other operational problems.  She forms the crucial link between QUOD and the SNODs.  After QUOD’s activity was paused during the first wave of COVID-19, she helped us with the mammoth task of restarting operations, updating SNOD training and finding ways around getting QUOD boxes delivered when so many personnel were working remotely between on-call shifts.

The elements of her job that she likes the most are the people and the research and innovation – ‘There is always wonderful new research going on such as the SIGNET trial and I find contributing to this work is really satisfying.’  She also said that her job is made very special by her colleagues, Hannah Tolley (OTDT Research Project Manager) and Emma Lawson (OTDT Innovation and Research Lead)). They work closely as a team linking in with other stakeholders to ensure that ODT research is facilitated safely and effectively. They also sit together alongside QUOD’s National Management Team.

I asked her if nursing was the career she had envisaged.  She laughed and said when she decided on nursing she was actually working at an agricultural college in Chelmsford, driving tractors and fruit picking.

Like all of us through the pandemic, Maggie has had to decamp her office to her home but she said she was one of many NHSBT staff who couldn’t bear sitting still when there were those working themselves to the bone on the front line so, she got herself deployed back to her original A&E department for six weeks.  She loved being back with her former colleagues.  I asked if she felt worried during that stint and she said ‘you just do what you have to do.’

Maggie Stevens is not all about work and loves an adventure in the great outdoors.  She has climbed Ben Nevis with colleagues for organ donation week and spent a couple of months travelling alone around Alaska where she saw whales and bears and met some wild and wonderful people that she will never forget.  She is also a fan of rock music and has tickets to see the Foo Fighters very soon!