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Abstract: SA-PO268

Human Primary Renal Proximal Tubule 3D Spheroid Model for Kidney Injury and Drug Discovery

Session Information

Category: Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

  • 2000 Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

Authors

  • Zhai, Yougang, Janssen Research and Development LLC, Raritan, New Jersey, United States
  • Kalyana-Sundaram, Shanker, Janssen Research and Development LLC, Raritan, New Jersey, United States
  • Pocai, Alessandro, Janssen Research and Development LLC, Raritan, New Jersey, United States
  • Wang, Lifeng, Janssen Research and Development LLC, Raritan, New Jersey, United States

Group or Team Name

  • CVMR-PH Discovery, Janssen Research & Development LLC.
Background

Renal proximal tubular epithelial cells (RPTECs) have a prominent role in maintaining kidney function by reabsorbing filtered nutritional substances, while allowing other substances to be excreted in the urine. RPTEC injury, cell death, and de-differentiation are mechanisms involved in kidney diseases including acute kidney injury (AKI). Human primary RPTECs isolated from normal human kidney tissue maintain the original phenotype in vitro for several passages and provide a translational advantage for studying nephron function and pathophysiology. Conventional two-dimensional culture methods frequently result in loss of tissue-specific RPTEC phenotypes partially due to changes of cell-cell communications and microenvironment. Hence, 3D models of RPTECs are being developed and used for drug discovery.

Methods

Primary hRPTECs were cultured in ULA spheroid plates for 4 days to form spheroids. Cellular injury was then induced by treatment with cisplatin or incubation in hypoxic chambers. Cytokine concentration, cell viability and apoptosis were measured by using commercial kits. RNA-seq were carried out, and pathway enrichment were analyzed by using Ingenuity Pathway Analysis software.

Results

Here we found that compared to 2D cultured cells, the hRPTEC 3D spheroid increased cisplatin induced injury responses and sensitivity with higher apoptosis and lower cell viability. Hypoxic condition induced significant increase of cell apoptosis and decrease of cell viability in hRPTEC spheroid. Transcriptomic analysis on the 3D hRPTEC hypoxia injury model identified similar pathway changes that found in in vivo in ischemic AKI models. Pathway analysis in this model supports a role of IL-17A in promoting hRPTEC inflammation and injury. In contrast, SIRT3 activator treatment protected hRPTEC from hypoxia injury likely by improving energy metabolism and mitochondria function.

Conclusion

3D injury models in primary hRPTEC spheroid are more sensitive to cisplatin injury than 2D cultures and pathway analysis revealed pathway changes consistent with a role of IL-17. The models provide a physiological and pathophysiological tool, to evaluate the function and pathophysiology of the kidney with higher translational relevance and can be leverage for fast screening and evaluation of potential therapeutic targets for kidney diseases.

Funding

  • Commercial Support – JNJ