Bioengineered kidney tubules efficiently excrete uremic toxins

Developers

J. Jansen, M. Fedecostante, M. J. Wilmer, J. G. Hoenderop, R. Masereeuw, etc.

Description of the technology

The technology was created in order to develop of a biotechnological platform for the removal of waste products (e.g. uremic toxins), often bound to proteins in plasma. This platform is a prerequisite to improve current treatment modalities for patients suffering from end stage renal disease.

The core of novel technology is in a newly designed bioengineered renal tubule capable of active uremic toxin secretion through the concerted functioning of essential renal transporters, namely, organic anion transporter-1 (OAT1), breast cancer resistance protein (BCRP) and multidrug resistance protein-4 (MRP4). Three-dimensional cell monolayer of human conditionally immortalized proximal tubule epithelial cells (ciPTEC), expressing endogenously a broad range of functional transporters associated with uremic toxin handling, was formed on membranes, consisting of biofunctionalized hollow fibers. The cells contained the temperature-sensitive mutant U19tsA58 of SV40 large T antigen (SV40T) and the essential catalytic subunit of human telomerase (hTERT), allowing the cells to proliferate at the permissive low temperature of 33°C and differentiate to mature PTEC at 37°C, and maintenance of telomere length preventing replicative senescence. This resulted in stable cell lines, which could be maintained over a long period of time and a valuable tool for studying renal clearance processes as required for the engineering of a bioartificial kidney.

The testing of new technology, which was carried out using a tailor made flow system, confirmed the secretory clearance of human serum albumin-bound uremic toxins, indoxyl sulfate and kynurenic acid, as well as albumin reabsorption across the bioengineered renal tubule.

Practical application

This technology proposes for clinical and experimental medicine the bioengineered kidney tubules, which can be perspective valuable method for treatment of patients suffering from end stage renal disease.

These functional bioengineered renal tubules are promising entities in renal replacement therapies and regenerative medicine, as well as in drug development programs.

The development of the technology led to following new important results. The functional bioengineered renal tubules were created that efficiently clear protein-bound anionic uremic toxins. Three dimensional, polarized, ciPTEC monolayers on biofunctionalized polyethersulfone hollow fiber membranes were developed. The essential role of in- and efflux transporters in the removal of uremic toxins was studied in systems of flat cell monolayers. The secretory clearance of human serum albumin-bound indoxyl sulfate and kynurenic acid, as well as albumin reabsorption, through novel structure, created in the frameworks of the engineering of a bioartificial kidney was confirmed.

Laboratories

• Department of Pharmacology and Toxicology, Radboud university medical center, Radboud Institute for Molecular Life Sciences, Nijmegen (The Netherlands)
• Physiology, Radboud university medical center, Radboud Institute for Molecular Life Sciences, Nijmegen (The Netherlands)
• Department of Pediatrics, Radboud university medical center, Nijmegen (The Netherlands)
• Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Utrecht (The Netherlands)

Links

Publications

• Jansen, J. et al. «Bioengineered kidney tubules efficiently excrete uremic toxins." 6 Scientific Reports, (2016): 26715.
• Jansen, J. et al. «Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations." 5 Sci Rep. (2015): 16702.
• Jansen, J. et al. «A morphological and functional comparison of proximal tubule cell lines established from human urine and kidney tissue." 323 Exp Cell Res. (2014): 87–99.
• Masereeuw, R. et al. «The kidney and uremic toxin removal: glomerulus or tubule?» 34 Semin Nephrol. (2014): 191–208.