Professor Stephen Marks, MD MSc MRCP DCH FRCPCH is Professor of Paediatric Nephrology and Transplantation at University College London Great Ormond Street Institute of Child Health and President of the British Association for Paediatric Nephrology. He is clinical lead for renal transplantation and Director of the National Institute for Health Research Great Ormond Street Hospital Clinical Research Facility at Great Ormond Street Hospital for Children NHS Foundation Trust. He has local, national and international roles including Chair of the Education Committee of the International Pediatric Transplant Association (IPTA) having also been an IPTA Councillor. His research continues to date in the fields of renal transplantation (including innovative drug trials concerning new anti-rejection therapies and assessment of children post-renal transplantation), systemic lupus erythematosus and vasculitis. He is on the editorial board for “Pediatric Nephrology” and “British Journal of Renal Medicine” and is associate editor for “Transplantation” and “Pediatric Transplantation”, which are the journals of The Transplantation Society (TTS) and the International Pediatric Transplant Association (IPTA), respectively.
Spatial transcriptomics in paediatric kidney transplant rejection
Barian Mohidin1, Stephen Marks1.
1UCL Great Ormond Street Institute of Child Health, London, United Kingdom
Background: Unfortunately, many children suffer kidney failure for a variety of reasons. Kidney transplantation provides the best outcomes for these children. However, kidney transplants are a finite resource and are unlikely to last beyond 25 years, meaning most children will need re-transplanting in the future. We aim to identify biomarkers that may help diagnose transplant rejection earlier and with greater sensitivity than monitoring serum creatinine.
Methods: First, we de-paraffinised and then stained five kidney transplant biopsy samples with allograft dysfunction with haematoxylin and eosin (H&E). We then used a proteinase digestion protocol to remove barriers that will impede RNA collection. The degraded samples were then stained using immunofluorescence techniques with a variety of antigen markers. The samples were then imaged using confocal microscopy.
Results: We confirmed a diagnosis of acute T-cell-mediated rejection (aTMR) with H&E and identified areas of borderline rejection (BR) in the same sample. We optimised our immunofluorescence staining protocol and decided on using CD45, Pan-CK, and Syto-13 stains for identifying leucocytes, tubular epithelial cells, and nuclei, respectively. We are now able to segment areas around leucocyte involvement in BR and aTMR.
Conclusions: We now plan on proceeding with spatial transcriptomics and measuring RNA transcripts in areas of BR and comparing it with areas of aTMR using Bruker GeoMx technology.