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Abstract: TH-PO444

UPARANT Succinate and N19004 Preserve Podocyte Injury by Adriamycin and Focal Segmental Glomerulosclerosis Sera: Drug Screening by an In Vitro Model of Glomerular Filtration Barrier

Session Information

Category: Glomerular Diseases

  • 1302 Glomerular Diseases: Immunology and Inflammation

Authors

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

Idiopathic focal segmental glomerular sclerosis (FSGS) accounts for up to 25-40 % of nephrotic syndrome with a frequent post-transplant recurrence due to a circulating permeability factor causing glomerular filtration impairment. In FSGS, an elevated level of soluble urokinase-type plasminogen activator receptor (suPAR) is implicated in podocytes' (PODO) alteration. We investigated the potential role of uPAR receptor inhibitors UPARANT succinate (US) and N19004 in preserving PODO structure and function.

Methods

Both UPARANT inhibitors were tested on ADRIA and FSGS-PODO injury models investigating the actin dynamic by IF actin and cofilin (CFL1) quantification. PODO differentiation and adhesion capacity were assessed by Wilms tumor 1 transcription factor (WT1), Integrin (ITG3), Nephrin (NPHS1) mRNA and protein expression. The functional barrier filtration capacity was assessed by a three-dimensional PODO-endothelial cell co-culture system.

Results

US and N19004 significantly protected PODO cytoskeleton with a well-orderly actin-filament and a foot processes effacement recovery after ADRIA-induced damage (vs ADRIA p=0.007, p<0.0001). Moreover, US or N19004 inhibited the ADRIA-induced actin-binding CFL1 phosphorylation (vs ADRIA p<0.0001 for both) already at 10nM. The activation of WT1, transcription factors of major PODO functional molecules such as ITG3 and NPHS1, was assessed. In addition, we found a NPHS1 perinuclear and cell processes distribution restoration upon drug treatment. Finally, uPAR receptor inhibitors recovered the functional barrier filtration capacity by the PODO-ENDO co-culture system upon activation by 3 FSGS sera patients. In the ADRIA-model only for N19004 at 100nM the recovery appears (vs CTRL p=0.02), conversely in the FSGS-model both US (vs CTRL p<0.05) and N19004 (vs CTRL p<0.02) improved the filtration functionality.

Conclusion

In conclusion, both drugs initiate a regain of the PODO plasticity and functionality, strongly encouraging future in-depth studies.