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Abstract: FR-PO702

In Vivo High-Content Zebrafish Screening Identifies Podocyte-Protective Drugs

Session Information

Category: Glomerular Diseases

  • 1304 Glomerular Diseases: Podocyte Biology

Authors

  • Schindler, Maximilian, Universitatsmedizin Greifswald, Greifswald, Mecklenburg-Vorpommern, Germany
  • Siegerist, Florian, Universitatsmedizin Greifswald, Greifswald, Mecklenburg-Vorpommern, Germany
  • Gehrig, Jochen, Acquifer Imaging GmbH, Heidelberg, Germany
  • Gul, Sheraz, Fraunhofer-ITMP, Hamburg, Germany
  • Endlich, Nicole, Universitatsmedizin Greifswald, Greifswald, Mecklenburg-Vorpommern, Germany

Group or Team Name

  • Endlich Lab
Background

Podocyte injury is the major cause of FSGS and no curative drugs are currently available. Therefore, identification of small molecules/compounds protecting podocytes is of high relevance. Since zebrafish larvae develop a size-selective filtration barrier within 4 days and we recently established a larval FSGS-like model, this organism is ideally suited for in vivo high-content screenings.

Methods

A transgenic screening strain expresses the nitroreductase and the fluorescent dye mCherry exclusively in podocytes. Additionally, a 78-kDa circulating eGFP-vitamin D-binding fusion protein is expressed. After incubation of larvae with 80 µM metronidazole (MTZ) for 24 hours, podocyte depletion and glomerular clearance of the eGFP fusion protein is induced. For the screening, larvae were co-treated with MTZ and compounds of a drug library at 4 days post fertilization (dpf) for 24 hours. After washout, the vascular eGFP as well as the podocyte mCherry was imaged at 5 and 6 dpf. The fluorescence ratios depict robust readouts for the degree of podocyte depletion and proteinuria.

Results

The screening of a drug library consisting of 138 compounds provided 9 potential hits with a protective effect on podocytes. These compounds significantly reduced the loss of vascular and/or podocyte fluorescence compared to the injury control group. Subsequent validation experiments with n=288 larvae per compound confirmed or rejected a protective effect of the drugs.

Conclusion

Our group established an in vivo high-content screening that identified potential drugs to treat FSGS.

Funding

  • Private Foundation Support