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Abstract: SA-PO577

A Novel 3D Cell Model That Mimics Renal Tubular Injury and Repair

Session Information

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Zhao, Shuan, Zhongshan hospital, Fudan University, Shanghai, China
  • Hu, Jiachang, Zhongshan hospital of Fudan University, Shanghai, SHANGHAI, China
  • Xu, Xialian, Zhongshan Hospital, Fudan University, Shanghai, China
Background

Acute kidney injury (AKI) is a major health problem with high morbidity and mortality. Despite decades of research, there is still no specific therapy for AKI. To develop targeted therapies to prevent or treat AKI, a basic prerequisite is a clear understanding of how cells repair injured kidney. When epithelial cells in the proximal portion of the nephron are damaged they rapidly proliferate to repair the damage to the kidney. Here, we developed a novel 3D culture model of AKI and, in a key insight, looked for genes that were involved in injury and repair.

Methods

A mosaic MDCK cysts were cultured in Matrigel and damaged at Day 7. During cysts repairing, total RNA of MDCK cells at indicated stages were extracted, purified and concentrated. After microarray analysis, genes that significantly expressed were selected, and verified in mouse ischemic reperfusion injury model at multiple time points by qRT-PCR.

Results

In our 3D culture system, cysts with monolayer of highly polarized cells were formed at Day6. Sectional cells in mosaic cyst could be induce apoptosis by oligomerization of a membrane localized, truncated form of caspase 8 and thereby cause damage to the cysts. Intriguingly, the remaining cells in the damaged cysts were able to re-enter the cell cycle, and restore the cysts to their pre-apoptotic state. Microarray analysis highlights a cascade of temporal-specific gene expression patterns related to tubular injury/repair. Seven genes that involved in cell junction and adhesion were selected and verified in animal AKI model. Three of those genes were significantly upregulated at 24h after IRI, two of those genes were dramatically reduced after IRI, and one gene’s expression has no change before 48h.

Conclusion

Using this 3D cell model of AKI, we have generated a list of gene candidates. Seven genes that related to cell junction and adhesion were identified, and expression patterns were similar between cell and mouse AKI model at multiple time points.

Funding

  • Government Support - Non-U.S.