Harries' lab: Team RNA
OUR ANIMAL FREE RESEARCH
The Harries team have interests in -omics approaches to the study of ageing and age-related disease processes in humans. Our work takes a genes-to-systems approach, ranging from ‘big data’ analyses to detailed individual molecular analysis of particular genes. Our work encompasses projects on cellular senescence and cell identity in the context of ageing and its diseases, and specifically the role of RNA processing and small RNA regulation in this context. We also house the Exeter Animal Free Research Centre of Excellence (ARC 2.0) funded by Animal Free Research UK.
HUMANISATION GUIDE
Our team has worked hard over the course of the Exeter Animal Free Research Centre of Excellence (ARC 2.0) to "humanise" many new cell models making their tissue culture processes animal-component free. We have collated our experience into a practical guide for human‑relevant tissue culture without the use of animal‑derived biomaterial. We hope this is of use to other animal-free researchers or researchers who want to consider a more human-relevant approach to their tissue culture.
AGEING & AGE-RELATED DISEASE
Dr Laura Sinclair is currently working on deciphering the role of sex hormones in ageing.
COVID-19
PhD student, Merlin Davies, developed a new animal free tool to detect COVID-19. You can learn more about his most recent published paper here.
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Merlin's PhD work consisted of the implementation and the discovery of new ways to introduce animal free science into the fields of ageing and immunology.
During the COVID-19 pandemic, Merlin developed a new PCR test for SARS-CoV-2 using completely non animal derived biomaterials. This test also detected active virus within patients, which meant results would inform patients whether they were still potentially infectious or not. Data from this study went on to help inform government decisions on pandemic management policy and isolation times.
Further to this, Merlin designed and utilised an animal derived biomaterial free panel of antibodies to identify the frequency of age-related markers in younger and older adult immune cells. These were used in a range of immunological methods such as flow cytometry and ELISAs. This data led to further understanding of the molecular and functional changes to immunity as humans age, especially in the context of SARS-CoV-2 infection.
DIABETES
Most recently Jodie Evans, a Graduate Research Assistant, was uncovering new approaches to treat diabetic kidney disease. This research involved humanising three different kidney cell lines (HK-2, HEK293 and HRPTEpC) so they can be cultured without the use of any animal-derived biomaterial. These cell lines were then be treated with the diabetomimetic stressors. Changes in HNRNPD gene and isoform expression (by qPCR analysis) and cellular viability, morphology, damage, and phenotypic markers were closely observed. Later, manipulation of HNRNPD isoform expression was performed, and cells were monitored for signs of resolution of such diabetomimetic stress.
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The idea for exploring diabetic kidney disease followed previous research projects led by Dr Nicky Jeffery and later followed up by Dr Sam Gould. Over a decade ago, Dr Nicky Jeffery assessed the causes of cell identity changes in pancreatic beta cells. To begin, Nicky humanised the EndoC-βH1 beta cell culture method. This allowed us to create a model for diabetes which more closely reflected human physiology. Through this research, we demonstrated that contrary to prior belief, loss of beta cell mass in diabetes derives not only from cell death, but also from changes in the identity of beta cells to other islet cell types induced by diabetes-related stresses, such as hyperglycaemia and altered lipid composition.
Following this discovery, Nicky led her second project where we were able to isolate the specific gene responsible for beta cell identity changes, the RNA binding protein hnRNPD (also known as AUF1). We identified this gene by isolating the altered ‘delta’ cell population by fluorescence-assisted cell sorting (FACS), using somatostatin as a marker. Following successful enrichment of the delta cell population, we carried out a whole transcriptome search for genes that demonstrated altered expression specifically in transdifferentiated ‘delta’ cells. Few coding genes demonstrated altered expression in somatostatin-positive cells, but of those that did, the most dysregulated gene was HNRNPD, which demonstrated a 2.7-fold reduction in expression compared to the somatostatin-negative cells in the culture.
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To begin our exploration into diabetic complications, Dr Sam Gould developed an animal free tissue-engineered retinal barrier model. This involved humanising both the Retinal Endothelial Cells (RECs) and Retinal Pigmented Epitheial Cells (RPEs). This model was developed to look at HNRNPD isoform expression in the retina, following diabetomimetic stimulation, with the view to gain a deeper understanding of the onset of diabetic retinopathy and ascertain whether HNRNPD isoform manipulation could act as a therapeutic option for this disease.
Pets of Team RNA
To show how much love Team RNA have for animals, we thought we would all share our beloved pets...
