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Abstract: SA-PO069

Urine-Derived Stem Cell Extracellular Vesicles Ameliorate Injury in a Human Kidney Organoid Model of Cisplatin-Induced Nephrotoxicity

Session Information

  • AKI: Mechanisms - III
    November 05, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
    Abstract Time: 10:00 AM - 12:00 PM

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Bejoy, Julie, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Welch, Richard C., Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Woodard, Lauren Elizabeth, Vanderbilt University Medical Center, Nashville, Tennessee, United States

Group or Team Name

  • Woodard Lab
Background

Urine-derived stem cells (USCs) are adult stem cells that originate from the human kidney and can be expanded. Extracellular vesicles (EVs) are mainly comprised of exosomes secreted from the endosomal system and microvesicles formed by direct outward budding. All cells produce EVs which transfer cellular cargo to specific target cells. Stem cell-derived EVs have clinical applications in disease treatment and diagnostics. Therefore, we also investigated USC-secreted EVs (USC-EVs) as a potential therapy to treat nephrotoxic acute kidney injury (AKI).

Methods

We tested USCs and USC-EVs in a human kidney organoid model of nephrotoxic AKI. We cultured human kidney organoids from induced pluripotent stem cells. We created a nephrotoxic injury model by adding 5 µm of the chemotherapeutic agent cisplatin to the kidney organoid culture for the first 24 h. For the next 48 h, we treated the injured organoids with either 5 x 104 USCs or 5 µg USC-EVs.

Results

First, we performed immunofluorescence for NEPHRIN, LTL, ECAD, and GATA3 to confirm differentiation of the organoids. In injured organoids treated with USCs, viability increased by MTT and LDH assay (p<0.001) and the expression of Kidney Injury Molecule-1 (KIM-1) appeared reduced by immunostaining. Treatment of injured organoids with USC-EVs also increased cell viability by MTT assay (p <0.0001) and decreased KIM1 release into the media by ELISA. USC-EVs lowered oxidative stress as we found increased superoxide dismutase (SOD) activity (p<0.05) and reduced Malondialdehyde (MDA) concentration (p<0.001). USC-EVs altered the levels of EGF (p <0.0001), IL-8 (p=0.0002), and IGF-15 (p<0.01).

Conclusion

USC and USC-EVs treatments increased viability of the cisplatin AKI organoids. USC-EVs reduced oxidative stress, KIM-1 and inflammatory cytokines. These results highlight the therapeutic effects of USC through release of USC-EVs in a human organoid model of nephrotoxic AKI, demonstrating the promise of future applications of USCs or USC-EVs for AKI treatment.

Funding

  • Veterans Affairs Support