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Abstract: FR-PO1071

Nephrotoxicity of Mixed Metals on Development/Injury in Zebrafish and Human Kidney Organoids

Session Information

Category: Health Maintenance, Nutrition, and Metabolism

  • 1500 Health Maintenance, Nutrition, and Metabolism

Authors

  • Emlet, David R., University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States
  • Merutka, Ilaria, Duke University Nicholas School of the Environment, Durham, North Carolina, United States
  • Price, Natalie, University of Pittsburgh School of Public Health, Pittsburgh, Pennsylvania, United States
  • Zimmerman, Zachary, University of Pittsburgh School of Public Health, Pittsburgh, Pennsylvania, United States
  • Turkington, Rachel E, University of Pittsburgh School of Public Health, Pittsburgh, Pennsylvania, United States
  • Eley, Moriah, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States
  • Gennings, Chris, Icahn School of Medicine at Mount Sinai, New York, New York, United States
  • Ho, Jacqueline, University of Pittsburgh Department of Pediatrics, Pittsburgh, Pennsylvania, United States
  • Sanders, Alison P., University of Pittsburgh School of Public Health, Pittsburgh, Pennsylvania, United States
  • Hukriede, Neil A., University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States
  • Jayasundara, Nishad, Duke University Nicholas School of the Environment, Durham, North Carolina, United States
Background

Toxic metal exposure can contribute to kidney disease. But the consequence of exposure to environmentally relevant nephrotoxic metal mixtures (NMM) at low levels are poorly understood. We tested toxicity of a combination of arsenic, cadmium and lead in human kidney organoids and zebrafish to examine changes in development, injury transcripts, and function.

Methods

NMM was defined from the National Health and Nutrition Examination Survey (NHANES) of urinary arsenite, cadmium, and lead in reproductive age women. Ratios were derived for total molar concentration, resulting in 35% arsenite, 36% cadmium, and 28% lead. Samples were exposed to a dose response from 1x-10,000xnM. We used a vascularized human kidney organoid system by mixing an inducible ETS translocation variant 2 human iPS cell line with a naïve line MANZ 2-2. Transcript levels after 6 days of NMM treatment were assessed in developing, differentiating, and mature organoids. Kidney function in zebrafish was assessed using a transgenic model which circulates a nano luciferase to assess proximal tubule function. We exposed developing fish to the NMM mixture and individual metals. Luminescence in surrounding water served as proxy for protein loss over a 24hour period.

Results

In mature organoids, NMM exposure demonstrated dose-responsive increases in expression of injury transcripts. In developing organoids, NMM suppressed transcripts essential for maturation at the 10x and 100x concentrations. Zebrafish results demonstrate that tubule function was impaired following exposure to NMM at the same concentrations. Strikingly, individual metal exposures did not produce any effect.

Conclusion

NMM exposures at environmentally relevant levels resulted in increased expression of injury genes and decreased developmental genes in a stage-specific manner in organoids, and decreased kidney function in zebrafish. Moreover, the clear interaction between metals to cause nephrotoxicity in the mixture but not in isolation indicate how important it is to assess multiple-stressor effects. These methods inform our population-based risk assessment strategy that coalesces translatable findings from kidney organoids and vertebrate model into a single framework.

Funding

  • NIDDK Support

Digital Object Identifier (DOI)