Kidney Week

Abstract: FR-PO1007

Plasma-Derived Extracellular Vesicles of Antibody-Mediated Allograft Rejection Kidney Transplant Patients Mediate Reprogramming of Human Podocytes: Role of SGLT2 Inhibitors

Session Information

• Transplantation: Basic
  October 25, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Transplantation

• 2101 Transplantation: Basic

Authors

• Castellano, Giuseppe, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy

Giuseppe Castellano, MD, PhD

• Armelloni, Silvia, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy

Silvia Armelloni, PhD

• Mattinzoli, Deborah, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy

Deborah Mattinzoli, PhD

• Li, Min, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy

Min Li, MD

• Ikehata, Masami, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy

Masami Ikehata, MD, PhD

• Bollati, Valentina, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy

Valentina Bollati, MD

• Abinti, Matteo, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy

Matteo Abinti, MD

• Podestà, Manuel Alfredo, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Lombardia, Italy

Manuel Alfredo Podestà, MD

• Alfieri, Carlo, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy

Carlo Alfieri, MD

Background

Extracellular vesicles (EVs), cell-derived particles, contribute to kidney diseases. Their role in antibody-mediated allograft rejection (AMR) is unclear. We aim to test their impact on human immortalized podocytes (PODO) and evaluate Dapagliflozin (Dapa) role as a pharmacological approach.

Methods

The study involved 28 Rtx patients (14 with AMR, 14 stable CTRL). PODO were exposed to plasma-EVs for 24h, then 100nM Dapa for 24h. Ultracentrifugation at 110,000g was used, followed by Nanosight counting and flow cytometry per MISEV2018. PKH26-EVs were used to examine PODO incorporation via Z-stack microscopy. RNA and protein were extracted for analysis, and cells were fixed for immunostaining.

Results

PODO incorporated and shared EVs. AMR-EVs increased CD9, a cell activation marker (p=0.028), induced a senescence-associated secretory phenotype in PODO (MCP1 p=0.041, TNFalpha p=0,046), and raised cytoskeletal rearrangement. They also increased the expression of EMT proteins like VIM (p=0.026), senescence P21 (p=0.04), inflammation CD44 (p=0.034), and decreased Klotho (p=0.002) (fig.1). Dapa incubation reduced PODO activation, decreasing P21 expression (p=0.039) and increasing Klotho (p=0.047), thus abrogating inflammaging (fig.2).

Conclusion

Our findings demonstrate that plasma AMR-EVs induce PODO reprogramming processes, comprising senescence, cytoskeleton rearrangement, secretion of inflammatory cytokines, decrease of nephroprotective protein Klotho and senescence increase. Dapa can efficiently counteract this pathogenic process.