Flow stimulates drug transport in a human kidney proximal tubule-on-a-chip independent of primary cilia

Highlights

• Albumin uptake and P-gp activity were enhanced in PTECs upon fluid shear stress.

• Fluid shear stress induced cell elongation corresponding with the direction of flow.

• Shear stress effects were not attributed to a primary cilium-related mechanism.

• No effect of fluid shear stress on cyclosporine A-induced cytotoxicity.

Abstract

Background

Kidney disease modeling and assessment of drug-induced kidney injury can be advanced using three-dimensional (3D) microfluidic models that recapitulate in vivo characteristics. Fluid shear stress (FSS) has been depicted as main modulator improving in vitro physiology in proximal tubule epithelial cells (PTECs). We aimed to elucidate the role of FSS and primary cilia on transport activity and morphology in PTECs.

Methods

Human conditionally immortalized PTEC (ciPTEC-parent) was cultured in a microfluidic 3D device, the OrganoPlate, under a physiological peak FSS of 2.0 dyne/cm² or low peak FSS of 0.5 dyne/cm². Upon a 9-day exposure to FSS, albumin-FITC uptake, activity of P-glycoprotein (P-gp) and multidrug resistance-associated proteins 2/4 (MRP2/4), cytotoxicity and cell morphology were determined.

Results

A primary cilium knock-out cell model, ciPTEC-KIF3α^−/−, was successfully established via CRISPR-Cas9 genome editing. Under physiological peak FSS, albumin-FITC uptake (p = .04) and P-gp efflux (p = .002) were increased as compared to low FSS. Remarkably, a higher albumin-FITC uptake (p = .03) and similar trends in activity of P-gp and MRP2/4 were observed in ciPTEC-KIF3α^−/−. FSS induced cell elongation corresponding with the direction of flow in both cell models, but had no effect on cyclosporine A-induced cytotoxicity.

Conclusions

FSS increased albumin uptake, P-gp efflux and cell elongation, but this was not attributed to a mechanosensitive mechanism related to primary cilia in PTECs, but likely to microvilli present at the apical membrane.

General significance

FSS-induced improvements in biological characteristics and activity in PTECs was not mediated through a primary cilium-related mechanism.