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Abstract: SA-PO1024

Expression Evaluation of CD2AP, ITGA3, and ITGB1 in Podocyte Cell Culture After Albumin Overload with and Without Puromycin-Aminoglycoside Damage

Session Information

Category: Glomerular Diseases

  • 1403 Podocyte Biology

Authors

  • Guaragna, Mara S., State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
  • Alves Troleze, José Ronaldo, State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
  • da Silva, Jackeline Pitonbeira, State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
  • Souza, Marcela Lopes de, State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
  • de Oliveira, Felipe Rodrigues, State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
  • Antonio, Heber da Silva, State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
  • dos Santos, Luana, State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
  • Mazzola, Taís Nitsch, State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
  • Torsoni, Adriana Souza, State University of Campinas, Faculty of Applied Sciences, Limeira, SP, Brazil
  • Belangero, Vera Maria Santoro, Universidade Estadual de Campinas Faculdade de Ciencias Medicas, Campinas, SP, Brazil
  • de Mello, Maricilda Palandi, State University of Campinas, Center for Molecular Biology and Genetic Engineering (CBMEG), Campinas, SP, Brazil
Background

Proteinuria is considered a prognostic marker for kidney disease and one of the main symptoms of podocytopathies. Several molecules participate in the maintenance of glomerular filtration barrier, including the CD2-associated protein (CD2AP) present in the slit diaphragm and the α3β1 integrin in the basal membrane of podocytes. This study aimed to evaluate the relative expression of CD2AP, ITGA3 and ITGB1 genes after overload of albumin in podocytes in vitro.

Methods

Conditionally immortalized human podocytes were exposed to progressive concentrations of albumin (0, 3, 20 and 40 mg/ml) for 24 hours under two conditions: with or without 12 hours of 15 ug/uL of puromycin aminoglycoside (PAN) treatment. We evaluated CD2AP, ITGA3 and ITGB1 genes relative expression through qPCR (n=3). The Friedman test was used to compare the different albumin exposures with and without PAN (p<0.05).

Results

The fold change of CD2AP, ITGA3 and ITGB1 genes are presented in Table 1. No significant difference was found among the albumin exposures, neither with nor without PAN (p>0.05).

Conclusion

In the condition here studied, albumin overload have not modulated CD2AP, ITGA3 and ITGB1 genes.

Fold change of CD2AP, ITGA3 and ITGB1 genes
PAN treatmentAlbumin exposure (mg/mL)Fold Change
CD2AP
Fold Change
ITGA3
Fold Change
ITGB1
No0111
30.0174181.152849610.938208375
200.0448711.4237300851.388398524
400.026110.5164051260.891691752
Yes0111
31.3862091.2411832051.258075685
201.7767011.7982578431.941487235
401.8961760.9824060171.587184907

Funding

  • Government Support – Non-U.S.