Abstract: PO0523
A New Physiological Model to Study Regulation of SLC26A6-Mediated Oxalate Transport in Mouse and Human Intestinal Tissue
Session Information
- Bone and Mineral Metabolism: Causes and Consequences
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Bone and Mineral Metabolism
- 401 Bone and Mineral Metabolism: Basic
Authors
- Schorr, Madeleine Josephine, Charite Universitatsmedizin Berlin, Berlin, Berlin, Germany
- Holthaus, David, Robert Koch Institut, Berlin, Berlin, Germany
- Fernandez Vallone, Valeria, Berlin Institute of Health, Berlin, Berlin, Germany
- Thomson, Robert Brent, Yale University School of Medicine, New Haven, Connecticut, United States
- Stachelscheid, Harald, Berlin Institute of Health, Berlin, Berlin, Germany
- Aronson, Peter S., Yale University School of Medicine, New Haven, Connecticut, United States
- Knauf, Felix, Charite Universitatsmedizin Berlin, Berlin, Berlin, Germany
Background
Intestinal organoids have great utility in studying stem-cell self-organizing properties. However, barrier and transport functions cannot be determined readily in three-dimensional (3D) cultures. We converted 3D intestinal organoids to two-dimensional monolayers (2D) and studied oxalate transport physiology via the oxalate transporter SLC26A6. Furthermore, we investigated the response of intestinal organoids to high oxalate concentrations.
Methods
Mouse and human adult stem cell-derived 3D culture systems were grown onto 2D monolayers. Cell differentiation was compared by gene expression and western blotting. Plasma membrane transport was examined in mouse and human monolayers with radioactively labeled substrates. Monolayers were exposed to soluble oxalate and cell death was measured by Caspase-3 activation and lactate dehydrogenase (LDH) release.
Results
We demonstrate that 2D intestinal monolayers maintained the gene expression profile of 3D organoids. Furthermore, murine and human intestinal organoids demonstrated high Cl-oxalate exchange transport activity that was 4,4-diisothiocyanostilbene-2,2-disulfonic acid (DIDS)-sensitive. Chloride-oxalate exchange was abrogated in murine organoids deficient for Slc26a6, resulting in intracellular oxalate accumulation, Caspase-3 activation and LDH release.
Conclusion
We conclude that 2D intestinal organoid cultures are suitable in vitro models to study oxalate transport from mice and humans. Using these models we demonstrate that Slc26a6-mediated chloride-oxalate exchange protects from intracellular oxalate accumulation and cell death.
Funding
- Government Support – Non-U.S.