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Abstract: PO2000

Prothrombin Modulates Podocyte Health and Function During Glomerular Proteinuria

Session Information

  • Podocyte Biology
    October 22, 2020 | Location: On-Demand
    Abstract Time: 10:00 AM - 12:00 PM

Category: Glomerular Diseases

  • 1204 Podocyte Biology

Authors

  • Waller, Amanda P., Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
  • Wolfgang, Katelyn, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
  • Wilkie, Tasha K., Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
  • Bhayana, Sagar, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
  • Davis, Kyle, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
  • Smoyer, William E., Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
  • Kerlin, Bryce A., Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States
Background

Ongoing podocyte injury is a known critical determinant of glomerular disease progression. Recent research suggests thrombin exacerbates in vitro podocyte injury, however, pharmacologic manipulation may cause both on- and off-target effects. Thus, the purpose of this study was to directly examine the effects of thrombin on glomerular proteinuria by manipulating its zymogen precursor, prothrombin (PT). We hypothesized that circulating PT would directly modulate both podocyte function and in situ survival in a rat model of nephrotic syndrome.

Methods

Puromycin aminonucleoside (PAN)-induced proteinuria was treated with: 1) PT antisense oligonucleotide to induce hypoprothrombinemia (LoPT), 2) Serial i.v. PT infusions to sustain hyperprothrombinemia (HiPT), or 3) sham (PAN only) controls (Con; n=12/group). Morning spot urine and citrated plasma were collected at day 10 post-PAN. Plasma PT activity was measured by chromogenic kit. Glomeruli were isolated from the kidney, dissociated into single-cell suspension and analyzed by flow cytometry following immunofluorescent antibody and TUNEL staining.

Results

Circulating plasma PT levels (Figure A) modulated proteinuria (B) such that it was significantly decreased in LoPT and increased in HiPT, compared to Con. LoPT also decreased in situ podocyte death (C), while HiPT increased in situ podocyte death, and resulted in fewer podocytes per glomerulus (D).

Conclusion

In conclusion, prothrombin modulates podocyte function (proteinuria) and survival (death and numbers) in the PAN model of glomerular proteinuria. Future studies should work to determine the prothrombinase mechanism that enables thrombin formation and signaling in the glomerulus and evaluate its potential as a novel therapeutic target to slow glomerular disease progression toward chronic kidney disease.

Funding

  • NIDDK Support