3D Proximal Tubule Tissues Recapitulate Key Aspects of Renal Physiology to Enable Nephrotoxicity Testing.
Due to its exposure to high concentrations of xenobiotics, the kidney proximal tubule is a primary site of nephrotoxicity and resulting attrition in the drug development pipeline. Current pre-clinical methods using 2D cell cultures and animal models are unable to fully recapitulate clinical drug responses due to limitedin vitrofunctional lifespan, or species-specific differences. Using Organovo's proprietary 3D bioprinting platform, we have developed a fully cellular humanin vitromodel of the proximal tubule interstitial interface comprising renal fibroblasts, endothelial cells, and primary human renal proximal tubule epithelial cells to enable more accurate prediction of tissue-level clinical outcomes. Histological characterization demonstrated formation of extensive microvascular networks supported by endogenous extracellular matrix deposition. The epithelial cells of the 3D proximal tubule tissues demonstrated tight junction formation and expression of renal uptake and efflux transporters; the polarized localization and function of P-gp and SGLT2 were confirmed. Treatment of 3D proximal tubule tissues with the nephrotoxin cisplatin induced loss of tissue viability and epithelial cells in a dose-dependent fashion, and cimetidine rescued these effects, confirming the role of the OCT2 transporter in cisplatin-induced nephrotoxicity. The tissues also demonstrated a fibrotic response to TGFβ as assessed by an increase in gene expression associated with human fibrosis and histological verification of excess extracellular matrix deposition. Together, these results suggest that the bioprinted 3D proximal tubule model can serve as a test bed for the mechanistic assessment of human nephrotoxicity and the development of pathogenic states involving epithelial-interstitial interactions, making them an important adjunct to animal studies.
Organovo, Inc. San Diego, CA, USA.
Shelby M
SM
King
Organovo, Inc. San Diego, CA, USA.
J William
JW
Higgins
Organovo, Inc. San Diego, CA, USA.
Celina R
CR
Nino
Organovo, Inc. San Diego, CA, USA.
Timothy R
TR
Smith
Organovo, Inc. San Diego, CA, USA.
Elizabeth H
EH
Paffenroth
Organovo, Inc. San Diego, CA, USA.
Casey E
CE
Fairbairn
Organovo, Inc. San Diego, CA, USA.
Abigail
A
Docuyanan
Ardea Biosciences Inc. San Diego, CA, USA.
Vishal D
VD
Shah
Organovo, Inc. San Diego, CA, USA.
Alice E
AE
Chen
Organovo, Inc. San Diego, CA, USA.
Sharon C
SC
Presnell
Organovo, Inc. San Diego, CA, USA.
Deborah G
DG
Nguyen
3D model; drug safety; nephrotoxicity; proximal tubule; renal transporters