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Abstract: TH-PO748

Dapagliflozin Exerts Senomorphic Effects on Adriamycin-Induced Podocyte Injury via Modulation of Complosome

Session Information

Category: Glomerular Diseases

  • 1403 Podocyte Biology

Authors

  • Castellano, Giuseppe, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy
  • Li, Min, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy
  • Mattinzoli, Deborah, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy
  • Armelloni, Silvia, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy
  • Ikehata, Masami, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy
  • Alfieri, Carlo, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy
Background

Recent clinical and experimental studies demonstrate that Sodium-glucose co-transporter 2 inhibitors (SGLT2i) exert reno- and cardiovascular protection independently of the hypoglycaemic effect. SGLT2i treatment rapidly reduces proteinuria preserving renal function in the long term. The mechanisms underlying the renal protection of SGLT2i are not fully understood. PODOs are the cornerstone of the glomerular filtration barrier, and podocytopathy is the principal cause of proteinuric glomerular diseases such as FSGS. In the present study, we attempt to verify if SGLT2i have a direct effect on Podocytes (PODO) after Adriamycin (ADR) injury investigating the underlying mechanisms.

Methods

Human-immortalized PODO and glomerular endothelial cells were used. Expression of SGLT2 Nephrin, Synaptopodin, p53, and p21WAF1 on PODO was investigated with immunofluorescence (IF), Western Blotting (WB), and quantitative PCR (qPCR). After ADR damage, the SGLT2i (Dapagliflozin, Dapa) at 100 nM was used to treat PODO. The PODOs’ actin cytoskeleton was evaluated. A PODO-endothelial cell co-culture device was used to test the albumin permeability (Palb). Intracellular Complement activation (Complosome) was analysed by confocal microscopy.

Results

PODOs express SGLT2 receptor. Dapa significantly decreased Palb provoked by ADR (ADR, 17%±5; ADR+Dapa, 2.5%±1, P<0.05) in vitro. Dapa repairs ADR-induced PODO injury by recovering of PODOs' F-actin, increasing Nephrin (mRNA: ADR, 0.04±0.01; ADR+Dapa, 0.29±0.13; P<0.05; protein: ADR, 0.21±0.02; ADR+Dapa, 0.57±0.3: P<0.05) and Synaptopodin (mRNA: ADR, 0.34±0.2; ADR+Dapa, 1.13±0.2; P<0.05; protein: ADR, 0.06±0.01; ADR+Dapa, 0.13±0.02; p<0.05). Importantly, Dapa down-regulates nuclear p53 (protein: ADR, 7.56; ADR+Dapa, 4.69) and p21WAF1 (ADR, 1.93; ADR+Dapa, 0.76). Moreover, we found that ADR actives complosome with a significant increase in intracellular C5a but not C3a. Dapa can inhibit intracellular C5a activation, shifting from the intracellular compartment to the outer cellular membrane.

Conclusion

SGLT2i exerts senomorphic effects restoring ADR-induced PODO inflammation and counteract Podo senescence. The underlying mechanisms could be the inhibition of Complosome activation at the C5a level.