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Abstract: SA-PO0140

Elucidation of Time-Resolved Kidney Metabolism in Health and Disease by Microfluidic-Coupled Metabolomics

Session Information

  • AKI: Mechanisms - 3
    November 08, 2025 | Location: Exhibit Hall, Convention Center
    Abstract Time: 10:00 AM - 12:00 PM

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Wulfmeyer, Vera Christine, Aarhus Universitet, Aarhus, Central Denmark Region , Denmark
  • Gernemann, Marlon, Aarhus Universitet, Aarhus, Central Denmark Region , Denmark
  • de la Rosa Siles, Xaiver, Aarhus Universitet, Aarhus, Central Denmark Region , Denmark
  • Rinschen, Markus M., Aarhus Universitet, Aarhus, Central Denmark Region , Denmark
Background

Measuring renal metabolism on the organ level is challenging, since metabolite signals in tissue and urine may reflect transport, metabolism, inter-organ communication, and incorporation into macromolecules - making it difficult to disentangle the kidneys’ specific role. A system allowing manipulation and time-resolved resolution in living kidney tissues would be necessary to overcome these limitations. We engineered a microfluidic system and workflow for the first time to control the input and measure the output of kidney tissues. Here, we further applied microfluidic-coupled metabolomics to tubules and glomeruli, examining time-resolved metabolic responses under both basal and stressor conditions in response to various stimuli.

Methods

Tubules and glomeruli were extracted from healthy mice (C57BL/6NTac) and rats (SD, Janvier Labs) according to established protocols. Tissues were subjected to basic buffer (electrolytes, hydrogen phosphates, AKG, gluconate, acetate, and glycine) or amino acid-containing buffers. Output was collected at 4°C, used for further metabolite extraction, and targeted high-sensitivity metabolomics was performed.

Results

The evaluation of time-resolved metabolism of tubules vs. glomeruli showed secretion of TCA metabolites (malic acid, succinic acid) and acetylated amino acids by tubules, and uncovered sex differences in tubular metabolism. We then evaluated the tubular oxidative stress response by the addition of H2O2 to one experimental tubule group under basic buffer conditions. H2O2 led to the release of lactic acid in the short term, and fumarate and malic acid in longer exposure, while impairing glucose secretion. Furthermore, metabolomics profiles demonstrated transport, metabolization, and time-dependent rebound under stressor conditions for a variety of substrates related to amino acid metabolism.

Conclusion

Microfluidic-coupled metabolomics with native tubules proved to be a valuable tool for the time-resolved evaluation of kidney metabolism. The findings are relevant in the context of acute kidney injury and support metabolic approaches to acute kidney disease. Further efforts will be made to characterize metabolism under stressor conditions and apply the system to a wide range of translational questions in health and disease.

Digital Object Identifier (DOI)