Kidney Week

Abstract: SA-PO645

Crotamine, a Cell Penetrating Peptide, Targeting Renal Proximal Tubular Epithelial Cells: A Potential Future for Kidney Gene Therapy

Session Information

• Pharmacology
  October 27, 2018 | Location: Exhibit Hall, San Diego Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

• 1700 Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

Authors

• Campeiro, Joana D., University Medical Center Groningen, São Paulo, Brazil

Joana D. Campeiro,

• Dam, Wendy, University Medical Center Groningen, São Paulo, Brazil

Wendy Dam,

• Lima, Sunamita Carvalho, Federal University of São Paulo, São Paulo, Brazil

Sunamita Carvalho Lima,

• Monte, Gabriela Guilherme, Federal University of São Paulo, São Paulo, Brazil

Gabriela Guilherme Monte,

• Oliveira, Lilian Caroline Gonçalves de, Federal University of São Paulo, São Paulo, Brazil

Lilian Caroline Gonçalves de Oliveira,

• Nering, Marcela Bego, Federal University of São Paulo, São Paulo, Brazil

Marcela Bego Nering,

• Viana, Gustavo Monteiro, Federal University of São Paulo, São Paulo, Brazil

Gustavo Monteiro Viana,

• Oliveira, Eduardo Brandt de, University of São Paulo, Ribeirão Preto, Brazil

Eduardo Brandt de Oliveira,

• van den Born, Jacob, University Medical Center Groningen, São Paulo, Brazil

Jacob van den Born,

• Hayashi, Mirian A.F., Federal University of São Paulo, São Paulo, Brazil

Mirian A.F. Hayashi,

Background

Activated proximal tubular epithelial cells (PTECs) play a crucial role in progressive tubulointerstitial fibrosis in native and transplanted kidneys. Active targeting of PTECs by non-viral gene delivery vectors such as cell penetrating peptides (CPPs) might be useful to influence the expression of genes in these cells, however most CPPs are not very cell type specific. Crotamine, isolated from the venom of rattlesnake, is characterized as CPP due to its ability to cross lipid cell membranes. Crotamine is also able to form complexes with DNA molecules and contains a nuclear retention motif, which helps to deliver DNA cargo into the nucleus enabling its use as a cell transfection agent. The ability of crotamine to internalize and carry molecules into cells is dependent on proteoglycans present on the cell membrane.

Methods

Crotamine was administered intraperitoneally to 14 week-old mice for 21 consecutive days. Main organs and body weight, food and water intake and renal function biomarkers were assessed. Fluorescently-labeled Cy3-crotamine was injected to track its localization throughout the kidney. Kidney was collected for immunohistochemistry assays. Crotamine binding, uptake and reporter gene expression was assessed in PTEC cell line.

Results

We demonstrated the safety of long-term crotamine administration and after injection into mice, crotamine passes the glomerular filter, is selectively taken up by PTECs, and subsequently localized into the nuclei of these cells. In vitro we could show that the binding and uptake of crotamine into PTEC cell line is mediated via the heparan sulfate side chains of syndecan-1, a major proteoglycan on these cells. In vitro we also showed efficient gene-delivery and reporter gene expression in this cell line.

Conclusion

This study shows the in vivo applicability of using crotamine as PTEC specific gene delivery nanocarrier and might be a prototypic example of the next generation kidney specific non-viral gene delivery vectors to modulate aberrant gene expression in PTECs in kidneys in order to slow down progressive tubulointerstitial fibrosis. Support: FAPESP/Capes.

Funding

• Government Support - Non-U.S.