Oral Presentation 12th International Meeting on AMPK 2023

Protection against cisplatin mediated renal tubular cell injury by the AMPK activator ATX-304 is not dependent on phosphorylation of acetyl-CoA carboxylase (#6)

Peter F Mount 1 2 , Marina Katerelos 1 , Kurt Gleich 1 , Kim Loh 3 , Bruce E Kemp 4 , David A Power 1 2
  1. Austin Health, Heidelberg, VIC, Australia
  2. Department of Medicine, University of Melbourne, Heidelberg, VIC, Australia
  3. Diabetes and Metabolic Disease, St Vincents Institute of Medical Research, Fitzroy, Vic, Australia
  4. St Vincents Institute of Medical Research, Fitzroy, VIC, Australia

Background: AMPK activation by metformin protects against cisplatin mediated injury to kidney cells, and this effect of metformin is suppressed by blocking AMPK signaling to acetyl-CoA carboxylase (ACC). ATX-304 is a direct AMPK activator that improves glucose homeostasis and microvascular perfusion in type 2 diabetes.

Methods: Primary cultures of kidney tubular epithelial cells (TECs) were pre-treated with ATX-304 before exposure to cisplatin (20 µM, 23-hrs). Cellular injury was studied in TECs from wild type (WT) C57Bl/6 control mice and ACC1/2 double knock-in (ACC1/2DKI) mice.

Results: In WT TECs, ATX-304 activated AMPK, with 20 µM for 4-hrs increasing AMPK-αThr172 phosphorylation 1.8-fold (P<0.05) and phosphorylation of ACC-Ser79 4.5-fold (P<0.01). By RT-PCR, ATX-304 (20 µM, 4-hrs) increased expression of peroxisome proliferator-activated receptor-γ coactivator (3.5+0.2-fold increase, P<0.0001) and sirtuin-3 (3.4+0.2-fold increase, P<0.0001). Pre-treatment with ATX-304 protected WT TECs 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), maintained cell viability (MTS assay), and reduced expression of the apoptosis marker cleaved caspase 3. In ACC1/2DKI TECs, greater reduction is cell viability (MTS assay) was observed (FC 0.57+0.03 vs 0.63+0.04 p=0.038). However, ATX-304 remained protective against cisplatin's effect on cell viability in the ACC1/2DKI TECs.

Conclusion: ATX-304 is protective against cisplatin mediated injury in TECs. Whilst blocking AMPK signaling to ACC increases the effect of cisplatin on cell viability, ATX-304 remained protective against injury in ACC1/2DKI TECs, indicating other AMPK actions contribute to this protective effect.