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Kidney Week

Abstract: PO0630

Lipid Metabolic Profiling of Ex Vivo Isolated Glomeruli as a Screening Platform for Modelling Glomerular Metabolic Dysfunction During Renal Disease

Session Information

  • CKD Mechanisms - 2
    October 22, 2020 | Location: On-Demand
    Abstract Time: 10:00 AM - 12:00 PM

Category: CKD (Non-Dialysis)

  • 2103 CKD (Non-Dialysis): Mechanisms

Authors

  • Romoli, Simone, Bioscience Renal, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Cambridge, United Kingdom
  • Krause, Fynn Niclas, University of Cambridge Department of Biochemistry, Cambridge, Cambridgeshire, United Kingdom
  • Griffin, Julian L., University of Cambridge Department of Biochemistry, Cambridge, Cambridgeshire, United Kingdom
  • Laerkegaard Hansen, Pernille B., Bioscience Renal, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Cambridge, United Kingdom
  • Woollard, Kevin, Bioscience Renal, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Cambridge, United Kingdom

Group or Team Name

  • Renal Bioscience ex-vivo glomerular team
Background

Dysregulated renal metabolism is a hallmark of loss of function in CKD. It is established that changes in tubular metabolism impact tubular functionality during progression of CKD. However, lipid metabolism in the glomerulus during CKD remain poorly described. Here, we use Isolated Glomeruli (IRG) to study lipid metabolism for metabolic drug discovery in kidney diseases.

Methods

Sprague Dawley rat glomeruli were isolated by differential sieving. We used disease inducers: Adriamycin 1uM (ADR) and AngiotensinII 100nM (AngII) for 24h. To probe metabolic activity, we used an LC-MS approach to quantify uptake and excretion rates of relevant metabolites in culture media, and to measure intracellular metabolites and lipids.

Results

We developed a new cultivation protocol for IRG, using organoids media and shaking platform to maximizing the biological activity. Metabolic and lipidomic profiles of IRG were monitored up to 150h. We saw significant metabolic activity for a wide range of metabolites: Uptake and excretion rates rapidly changed during the first 24h of culture, after which they declined. Metabolic rates for glutamine, glutamate and alanine were comparatively stable. Following treatment with AngII or ADR glomeruli exhibited metabolic changes after 24h: AngII reduced asparagine uptake, and induced trends towards lower substrate uptake consistent with reduced metabolic activity. Both ADR and AngII perturbated intracellular metabolite levels: nucleosides adenosine (-159%) and inosine (-171%), increases in 1,3-BPG (+194%), and changes in NAD (+209%), which suggest alterations in pentose phosphate pathway. Multivariate analysis revealed differentially responding lipid clusters: specifically, significant abundance and saturation ratio increases of intracellular FFA, including stearate (+34%), oleate (+102%) and arachidonate (+107%), as well as the depletion of several phosphatidylcholine and phosphoethanolamine species following AngII, which have been implicated as renal injury markers and/or relevant to renal injury protection.

Conclusion

Our results show alterations in lipid metabolism after IRG stimulation with AngII and ADR after 24h. We propose IRG/lipid metabolome as a novel platform/tool for understanding lipid signalling and improving CKD drug discovery.

Funding

  • Commercial Support