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!

COVID BioArchive Update

Since March 2020 the QUOD team has been supporting the national effort in the fight against COVID-19. QUOD’s extensive biobanking expertise, infrastructure and personnel have served to set up the NHSBT Oxford COVID BioArchive (COBA).  Over 68,000 blood samples from convalescent plasma donors who have recovered from COVID-19 have been collected and processed as part of the NHSBT Convalescent Plasma programme. In addition, since September COBA has received over 18,000 samples collected from recipients of convalescent plasma and monoclonal antibody treatment in the international RECOVERY trial.

These samples were used by a number of research groups, NHSBT and Public Health England in a variety of projects, including assessment of novel COVID-19 tests, characterisation of COVID-19 antibody function, and analysis of the efficacy of convalescent plasma treatment. With this COVID BioArchive, we have been able to establish a robust and sustainable resource for future validation and research helping us to gain better insight into COVID-19 and anticipate targeted intervention.

You may have heard that on 15th January randomisations of patients into convalescent plasma was paused following disappointing interim results that showed no evidence that convalescent plasma has an overall benefit on patient outcomes in moderately ill people.  Work to search for evidence of benefit in subgroups before organ damage and hospitalisation occurs is now under consideration.

Given the quantity of samples now available, it has been decided that collection of further plasma samples will not be resumed.  The news may seem discouraging, however an important scientific question has been answered.  In terms of the scale and volume of the operation this was a considerable undertaking that has proved that the QUOD infrastructure in close collaboration with the NHSBT’s Blood Service has the capacity and versatility to help and meet such an important demand.

Over 250,000 samples are currently stored in the COVID BioArchive to be used for dedicated research questions and validation of novel tests. Applications are welcome from national and international research groups or health care authorities.

BTS & NHSBT Congress 2021 Roundup

by Rebecca Vaughan

This year, as has become our new norm, the BTS Congress 2021 was hosted using an online platform, allowing delegates to experience interactive plenary sessions and parallel sessions similar to when attending the conference.  However unique to the online experience was the opportunity to access content ‘on-demand’, including the additional option to listen to pre-recorded abstract talks, at a time that suits you.  Although, many were saddened by the lost opportunity to meet with colleagues and friends, the commitment from BTS to provide an opportunity for clinicians and researchers to gather and share their research from the last 12-months was an absolute success and credit to the organisers. 

For this conference, I submitted an abstract titled ‘Brain death specific glomerular matrix degradation profiles are associated with long-term graft dysfunction in kidney transplant’ on behalf of our teams at NHSBT and the Oxford Transplant research lab in University of Oxford.  This abstract was selected for presentation as part of the ‘Medawar Award Session’, where my abstract and presentation was considered for the award along with three other researchers who also submitted abstracts for basic science in transplantation. I feel incredibly fortunate to have had our abstract considered for the Medawar award, and was excited to highlight how we have utilised invaluable samples from the QUOD biobank.  Our research utilised 60 kidney biopsies from the QUOD biobank from both DBD and DCD donors.  Our work described a DBD specific degradation profile for cytoskeletal proteins in donor kidneys that linked to suboptimal 12month posttransplant function.  Our research was considered novel in that it indicates that protein degradation may affect donor kidneys and post-transplant function. Preparing for my live presentation was also a memorable experience, involving many rehearsals with my supervisors Dr Kaisar and Prof Ploeg and colleagues, many adjustments and changes before the final live talk.

Our group also had a second opportunity to present our research work involving the analysis of  QUOD samples. A second abstract titled ‘Protein profiles in deceased donor kidneys associated with 12-Month post-transplant kidney function’ was accepted as a pre-recorded talk, presented by our colleague Priyanka Joshi.  This study utilised 185 kidney biopsies from the QUOD biobank for proteomic analysis, comparing DBD and DCD donors with differing functional outcomes 12mth posttransplantation.  This study showed that protein markers in donor kidneys associate to posttransplant outcomes and indicate which biological pathways may play a role in determining the donor kidney quality.

Our research work has highlighted the value of the QUOD samples; based on the collection protocols, these samples provide us with confidence regarding sample quality and collection continuity.  Importantly, our sample selection and analysis is linked with the clinical and demographic metadata that we receive from the National Transplant Registry from NHSBT. The combination of the selection of QUOD samples and donor and recipient metadata makes a unique research resource.  

The overarching focus of the BTS congress was the impact of COVID-19 on transplantation, with NHSBT statistics team highlighting changes in trends of donation and transplantation due to the pandemic.  An invited keynote speaker focused on ‘resilience’  encouraging people to reflect on their own resilience and how they have adjusted to the inevitable changes that have come along with a global pandemic, whilst looking to the future with reasonable caution but also excitement.  Several talks also touched on emerging perfusion technologies and the improvements they may have on outcomes in the future, and evaluating single centre experiences.

Overall the BTS Congress 2021 was a great success despite the mammoth task of moving the conference to an online platform.  The opportunity to bring together scientists and clinicians to discuss research, trends and experiences in transplantation is essential to moving the field forward.  The conference provided an excellent opportunity to showcase research work, however the opportunity to interact with each other; ask questions, discuss and debate ideas was lacking and greatly missed.  The representation of QUOD, NHSBT and the University of Oxford was strong, and hopefully more researchers were inspired to consider QUOD samples for their ongoing work.

Congratulations to Charlotte Brown (Wales Kidney Research Unit) and Maria Ibrahim (NHSBT Bristol) who were awarded the 2021 Medawar Medals at the congress. Click here to read about their research.

Increasing the Number of Organs Available for Research (INOAR)

Until recently, only organs removed for transplant, but subsequently not transplanted were available to researchers.  Thanks to a collaboration between QUOD and Newcastle University, in partnership with NHS Blood and Transplant, new arrangements have been implemented that will allow hearts, lungs and pancreases which are unsuitable for transplantation to be retrieved for research purposes. This will greatly facilitate ongoing research into developing ways more donated organs can be converted into successful life-saving transplants.

Clare Denison, Lead Specialist – Innovation and Research ODT at NHS Blood and Transplant, said: “This is a significant moment for our organisation and researchers across the country. INOAR will change the face of transplantation and ultimately improve patient outcomes and quality of life in the future.”

This development is particularly exciting for diabetes researchers. Almost 4 million people in the UK are living with diabetes. The condition occurs when the pancreas doesn’t produce enough insulin, or when it can’t produce any at all, which leads to blood glucose levels being too high. Until now, the pancreas has not been removed, or even sampled following the death of people with diabetes during organ donation for transplantation.  Research into the mechanisms preventing normal pancreatic insulin production in diabetes has therefore been limited to the small number of post-mortem samples currently stored in the UK.

Dr Elizabeth Robertson, Director of Research at Diabetes UK, said: “This is an exciting step forward that we hope will rapidly advance our understanding of the causes and progression of diabetes … world-class scientists will now have vital access to pancreatic tissue, propelling our knowledge of diabetes forward and bringing us a step closer to a world where diabetes can do no harm.”

Spotlight on a QUOD Colleague: Dr Maria Kaisar

by Susan Patchett

This time we will be getting to know Dr Maria Kaisar who is a scientist leading valuable research to identify biomarkers to better assess the quality of deceased donor organs.  She explained that biomarkers can be proteins or genes that can be measured during organ donation to indicate the risk of poor function or even failure after transplantation.  Her work also investigates biological processes that may be altered during organ donation and impact on effective organ function.  She says that the more that can be learned about the changes in biological mechanisms, the closer we come to designing new therapies to repair organs and make transplants last longer.

Dr Kaisar says she feels privileged to have been part of the QUOD team since its creation in 2012.  She recalls the strong collaborative spirit of the clinical teams (NORs, SNODs), academic partners and scientists across a host of institutions including NHSBT and transplant centres who all contributed to the development of the QUOD project.  She has watched QUOD evolve into the platform it has become today and takes great pleasure in seeing QUOD samples supporting so much exciting research both nationally and internationally.

Maria has also been working on the COMPARE Trial that showed that the addition of oxygen during hypothermic machine perfusion of older DCD kidneys improved transplant outcomes.  COMPARE is a component of the Consortium for Organ Preservation in Europe (COPE).

Every day is different for Maria as she spends her time in meetings, designing new experiments, analysing data and drafting grant applications and papers.  She splits her time between her lab at the NHSBT Filton Blood Centre in Bristol, where she has built a small team, and at the University of Oxford working alongside colleagues in the QUOD/Ploeg Research lab at the Oxford Blood Centre. She also holds responsibilities as Vice-Chair of ESOT’s (European Society of Transplantation) Basic Science committee. Working with colleagues from across the United Kingdom and further afield in the Netherlands, Austria, Denmark, France and Belgium has provided valuable friendships that extend beyond scientific interest.

Maria is a passionate advocate of the Women in Science initiative and diversity in science.  She says ‘We must rethink scientific careers to support career progression of scientists to senior leadership roles whilst caring for families – COVID has taught us that we can be productive in many ways and incorporate flexibility achieving a healthy work life balance. We also should aim to be more ethnically diverse as we can only gain valuable insights and perspectives’. The most significant obstacle faced by scientists today, she says, is the lack of long term, secure funding.  COVID has shown that investment in science is essential to meet the changing medical challenges we are faced with. 

Dr Kaisar’s journey in science began with her interest in biology at school, though she was also keen on philosophy, ‘very Greek!’ she laughs.  With a BSc in Chemistry from the University of Athens, she took up a one-year internship that became a six-year research role at Imperial College School of Medicine. After taking some time out to start a family, she returned to work for NHSBT and developed a strong interest in organ transplantation that led to joining Professor Ploeg’s group and completing a DPhil at the University of Oxford.

When Maria isn’t pondering science she loves holidays with her family, picnics, long walks and cooking.  She also loves live music and going to concerts and gigs which she hope to be able to do again soon.

Spotlight on a QUOD Colleague: Dr Hannah McGivern

by Susan Patchett

As part of our new series of spotlights on QUOD colleagues, I caught up with QUOD’s Tissue Handling Technician, Dr Hannah McGivern this afternoon.

Hannah takes receipt of and processes donor tissue samples; that is to say biopsies from both deceased and living donor organs, sent to us by QUOD hospitals up and down the country.  These samples are then stored in the biobank ready for researchers to apply for access to.

Hannah finds her work rewarding and relished the opportunity to learn the raft of new skills in soft tissue processing. She says that she feels privileged to collect samples in theatre for the Oxford Transplant Biobank (OTB), and to play a small part in this important moment in the lives of living donors and recipients. 

2020 has, of course, been a difficult year and the sudden and dramatic impact of the pandemic has been felt by everyone.  Hannah has also been working alongside QUOD colleagues to process plasma samples from individuals who have recovered from COVID-19 for the Convalescent Plasma Programme and for the new COVID BioArchive (COBA).  The pressure has been high in terms of the sheer volume of samples; the record was 300 samples processed in just one day!  Hannah feels honoured to be contributing to work that has the potential to have an important impact on the world; and on the future of biological science; ‘This is what every scientist dreams of.’  You can read more about COBA in our latest newsletter.

Hannah began working with us just over a year ago, during the final stages of her PhD at Cranfield University.  Her studies focussed on the structure and mechanical properties of the human skeleton.  She looked, in particular, at the ribs and clavicle and how their condition changes as we get older. Her work showed that our mechanical strength peaks in our mid-thirties before deteriorating.

Hannah’s interest in science began at the tender age of seven when her grandfather, an engineer in the Royal Air Force, described yeast as ‘little animals’.  At around eleven, her love of science was set in concrete when, at school, she came across a crime scene investigation kit which taught pupils the basic principles of forensic science, including how to take fingerprints. With her appetite whetted, she went on to study for a BSc in Archaeology with Forensic Science at the University of Exeter, followed by an MSc in Forensic Osteology at Bournemouth.

She has participated in archaeological digs as far afield as the USA and Iceland.  At Cranfield, she assisted with the cataloguing of human remains from a dig, colloquially dubbed Rat Island (Burrow Island), a tiny peninsula that juts into Portsmouth Harbour.  It is thought that the skeletons found there could be prisoners from the late 18th and early 19th centuries.  It was revealed that one may have undergone surgical experimentation, post-mortem, as the top of their skull had been removed.  The dig featured on ‘Digging for Britain’ a production for the BBC presented by Professor Alice Roberts.  Hannah is fascinated about what can be learned about the past from skeletal remains.  If you’d like to know more about this project click here.

Hannah is passionate about outreach work and contributes wholeheartedly to public engagement for QUOD.  In addition, she was selected to participate in Soapbox Science, an interactive outreach event to promote the work of female scientists. She is also a volunteer STEM (Science, Technology, Engineering and Maths) Ambassador going into schools to raise awareness of the careers available to women.  Having grown up in an area of the country  where the uptake of higher education is lower than much of the country, she strives to change preconceptions about female scientists and inspire young women to embark on careers they may not have deemed possible.  She is an avid supporter of the ‘Women in Science’ initiative and points out that QUOD, being staffed predominantly by women, is a perfect example.  You can read her blog by clicking here.

Though it is not all work, work, work for Hannah; she has a passion for literature and film, was an avid fan of Top Gear growing up and when she can, enjoys Latin and ballroom dancing.

Meet the Team: Sheba Ziyenge, Data Analyst and Coordinator

In September, QUOD welcomed our new Data Analyst and Coordinator Sheba Ziyenge. Sheba has kindly agreed to share a bit more about herself and her role at QUOD:

What were you doing most recently before joining QUOD?
I was doing my Master’s degree in Cognitive Neuroscience in Barcelona.

What interested you about working with QUOD?
From my prior experiences, I knew I wanted to transition to using my data management, statistical and analysis skills in scientific research. QUOD stood out to me for their fantastic work in supporting contemporary transplantation research.

What does your role in QUOD involve?
My main duties include the management and development of QUOD databases, the extraction and allocation of biobank samples to research projects, supporting researchers with analysis and reporting. I also produce periodic statistics about QUOD activities for stakeholders and collaborate with NHSBT to process data relevant for the research projects.

Lightning round time. Tea or coffee?
Coffee in the morning, tea in the afternoon!

Favourite place in Oxford?
I love walking by the river up to Christ Church Meadows, it’s especially nice in the summer.

Ideal holiday destination?
I love travelling so this is a hard one! My favourite place I’ve been to so far is Mexico, and I have my eyes set on New Zealand and Hawaii for my next big trips!

Primary COVID-19 lockdown survival method?
Enjoying the little things such as burning scented candles, listening to nice music and looking after my plants.

Finally, what kinds of inquiries should people bring to you, and how best can they reach you?
I can be reached at sheba.ziyenge@nds.ox.ac.uk, I’ll be happy to help with any QUOD data related queries.