Abstract: TH-PO120
The AMPK Activator ATX-304 Reprograms Tubular Cell Metabolism to Protect Against Cisplatin-Induced AKI
Session Information
- AKI: Mechanisms - I
November 02, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Mount, Peter F., Austin Health, Heidelberg, Victoria, Australia
- Katerelos, Marina, Austin Health, Heidelberg, Victoria, Australia
- Gleich, Kurt, Austin Health, Heidelberg, Victoria, Australia
- Harley, Geoffrey, Austin Health, Heidelberg, Victoria, Australia
- Lee, Mardiana, Austin Health, Heidelberg, Victoria, Australia
- Loh, Kim, St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia
- Oakhill, Jonathan S., St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia
- Narayana, Vinod Kumar, The University of Melbourne Bio21 Molecular Science and Biotechnology Institute, Parkville, Victoria, Australia
- De Souza, David P., The University of Melbourne Bio21 Molecular Science and Biotechnology Institute, Parkville, Victoria, Australia
- Laskowski, Adrienne, Monash University Central Clinical School, Melbourne, Victoria, Australia
- Coughlan, Melinda T., Monash University Central Clinical School, Melbourne, Victoria, Australia
- Kemp, Bruce E., St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia
- Power, David A., Austin Health, Heidelberg, Victoria, Australia
Background
Acute kidney injury (AKI) is characterized by widespread disruption of energy metabolism pathways. Targeting the metabolic regulator AMP-activated protein kinase (AMPK) is a potential strategy for addressing the problem of AKI. ATX-304 is a novel PAN-AMPK activator with reported benefits in models of diabetes, obesity and aging.
Methods
C57Bl/6 mice were fed chow containing ATX-304 (1mg/gm) versus control chow for 7-days prior to cisplatin-induced AKI (CI-AKI). Primary cultures of tubular epithelial cells (TECs) were pre-treated with ATX-304 (20 µM, 4-hrs), prior to exposure to cisplatin (20 µM, 23-hours).
Results
ATX-304 protected against CI-AKI as measured by serum creatinine at 48-hours (control 0.05+0.03 mM vs ATX-304 0.02+0.01 mM, P=0.004), western blot for neutrophil gelatinase-associated lipocalin (NGAL) (control 3.3+1.8-fold vs ATX-304 1.2+0.55-fold, P=0.002), RT-PCR for NGAL (control 99.6+22.8-fold vs ATX-304 35.2+5.9-fold, P=0.006), monocyte chemoattractant protein-1 (control 18.7+5.1-fold vs ATX-304 10.4+1.3-fold, P=0.048) and receptor-interating protein kinase 3 (CI-AKI control 6.9+2.4-fold vs CI-AKI ATX-304 4.9+1.3-fold, P=0.02), and histological injury score (control 3.5+0.59 vs ATX-304 2.7+0.74, P=0.03). In TECs, pre-treatment with ATX-304 protected from cisplatin mediated cell death, measured by lactate dehydrogenase release assay (control 27.1+4.2% lysis vs ATX304 -0.6+2.5% lysis, p<0.0001), and maintained cell viability in the presence of cisplatin, as measured by MTS assay. ATX-304 protection against cisplatin was lost in AMPK-null murine embryonic fibroblasts. In TECs metabolomic analysis revealed that ATX-304 (20 µM, 4-hrs) altered the level of 66/126 detected metabolites, including changes in fatty acids, tricarboxylic acid cycle metabolites, and amino acids. Energy metabolism studies of live TECs using the XFe96 Seahorse analyser found ATX-304 increased basal oxygen consumption rate by 38%, whereas maximal respiration was unchanged.
Conclusion
The mechanism of ATX-304 protection against cisplatin injury is AMPK- dependent and involves widespread metabolic reprogramming. AMPK activation by ATX-304 is a promising therapeutic strategy for AKI.
Funding
- Commercial Support – Amplifier Therapeutics (Betagenon)