Grantee Research Project Results

2016 Progress Report: Organotypic Model of Human Kidney as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials

EPA Grant Number: R835738C002
Subproject: this is subproject number 002 , established and managed by the Center Director under grant R835738
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: Predictive Toxicology Center for Organotypic Cultures and Assessment of AOPs for Engineered Nanomaterials
Center Director: Faustman, Elaine
Title: Organotypic Model of Human Kidney as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
Investigators: Kelly, Edward J.
Institution: University of Washington
EPA Project Officer: Aja, Hayley
Project Period: December 1, 2014 through November 30, 2018 (Extended to November 30, 2019)
Project Period Covered by this Report: December 1, 2015 through November 30,2016
RFA: Organotypic Culture Models for Predictive Toxicology Center (2013) RFA Text | Recipients Lists
Research Category: Chemical Safety for Sustainability

Objective:

One of the primary objectives of our project is to design, implement and test a tissue-engineered human kidney microphysiological system and to evaluate the response of exposure to engineered nanomaterials (ENMs). To this end, we evaluated the toxicological effects of two forms of silver nanoparticles (AgNPs) with an organotypic microfluidic device that utilizes the Nortis™ microphysiological (MPS) system, which accurately reflects human renal physiology with the culturing of primary human proximal tubule epithelial cells (PTEC) in a physiologically relevant 3-D configuration and an appropriately scaled lumenal flow rate.

Progress Summary:

Silver Nanoparticles

We exposed PTECS to either 20 μg/mL of 20 nm PVP Ag nanoparticles plus an albumin/lipid complex, the modified albumin/lipid complex media alone or our standard PTEC cell culture media (negative control) for 48 hours and collected effluents for the presence of KIM-1. While the supplementation of albumin and lipids to the PVP nanoparticle stock solution prior to addition to the cell culture media prevented immediate aggregation there were visible particle aggregates in the MPS devices after 48 hours of exposure. More importantly was that the addition of the albumin/lipid complex to the culture media caused as much KIM-1 release as the treatment of PVP AgNP dissolved in the albumin/lipid complex.

PTEC treated with citrate or PVP-coated AgNP lacked any significant toxicity as measured by ICC methods to detect HO-1 and KIM-1 and by measurements of KIM-1 in the effluents of the devices. Additionally, there was no evidence of loss of viability PTEC after AgNP exposure.

Quantum Dots

Quantum dots (QD) with a CdSe/ZnS core and a net positive charge were chosen for PTEC exposure experiments due to their nephrotoxic potential with the cadmium/selenium core and for their solubility capabilities. As with previous nanoparticle assessments, we conducted dose-range finding experiments with QDs exposures in a 96-well format to assure solubility as we evaluated cell morphology, cell viability/health and mitochondrial function to select an EC50 concentration that was later used to set MPS exposure concentrations.

Future Activities:

We are presently planning experiments to evaluate the toxicity of CdSe/ZnS core/shell quantum dots coated with neutral (polyethylene glycol) surface charges in an MPS format. These quantum dots are commonly used in the electronics industry, contain the toxic heavy metals Cd and Se and have already been shown to be toxic in other cell types. We have already demonstrated that we can detect PTEC toxicity with increased HO-1 and KIM-1 immunofluorescence in MPS after 48 hour exposure to 25 µM CdCl2 (Adler, Ramm et al. 2015). The results with CdCl2 are part of the control toxicants used to evaluate the MPS platform. Anticipated additional analyses include the assessment of cellular ultrastructures by electron microscopy, Adverse Outcome Pathway (AOP) determinations using RNA-seq transcriptomic analysis and immunocytochemistry (ICC) fluorescent imaging for KIM-1 and HO-1 and MPS effluent analysis for KIM-1 and ICP/MS measurements of effluents. Other response ICC markers that will be evaluated include: SGLT2 and E-cadherin while additional effluent markers include interleukin 18 and Neutrophil gelatinase-associated lipocalin (NGAL).

Journal Articles on this Report : 5 Displayed | Download in RIS Format

Publications Views
Other subproject views:	All 64 publications	17 publications in selected types	All 16 journal articles
Other center views:	All 150 publications	50 publications in selected types	All 49 journal articles

Publications
Type	Citation	Sub Project	Document Sources
Journal Article	Chang S-Y, Weber EJ, Van Ness KP, Eaton DL, Kelly EJ. Liver and kidney on chips: microphysiological models to understand transporter function. Clinical Pharmacology & Therapeutics 2016;100(5):464-478.	R835738 (2016) R835738 (2017) R835738C002 (2016) R835738C003 (2016) R835738C003 (2017) R835738C005 (2017)	Full-text from PubMed Abstract from PubMed Associated PubMed link Abstract: Wiley-Abstract Exit Other: ResearchGate-Abstract Exit
Journal Article	Chang S-Y, Voellinger JL, Van Ness KP, Chapron B, Shaffer RM, Neumann T, White CC, Kavanagh TJ, Kelly EJ, Eaton DL. Characterization of rat or human hepatocytes cultured in microphysiological systems (MPS) to identify hepatotoxicity. Toxicology In Vitro 2017;40:170-183.	R835738 (2016) R835738 (2017) R835738C002 (2016) R835738C002 (2017) R835738C003 (2016) R835738C003 (2017)	Abstract from PubMed Full-text: ScienceDirect-Full Text HTML Exit Abstract: ScienceDirect-Abstract Exit Other: ScienceDirect-Full Text PDF Exit
Journal Article	Vernetti L, Gough A, Baetz N, Blutt S, Broughman JR, Brown JA, Foulke-Abel J, Hasan N, In J, Kelly E, Kovbasnjuk O, Repper J, Senutovitch N, Stabb J, Yeung C, Zachos NC, Donowitz M, Estes M, Himmelfarb J, Truskey G, Wikswo JP, Taylor DL. Functional coupling of human microphysiology systems: intestine, liver, kidney proximal tubule, blood-brain barrier and skeletal muscle. Scientific Reports 2017;7:42296 (14 pp.).	R835738 (2017) R835738C002 (2016) R835736C004 (2017)	Full-text from PubMed Abstract from PubMed Associated PubMed link Full-text: Scientific Reports-Full Text HTML Exit Abstract: Scientific Reports-Abstract Exit Other: Scientific Reports-Full Text PDF Exit
Journal Article	Vernetti L, Gough A, Baetz N, Blutt S, Broughman JR, Brown JA, Foulke-Abel J, Hasan N, In J, Kelly E, Kovbasnjuk O, Repper J, Senutovitch N, Stabb J, Yeung C, Zachos NC, Donowitz M Estes M, Himmelfarb J, Truskey G, Wikswo JP, Taylor DL. Corrigendum: Functional coupling of human microphysiology systems: intestine, liver, kidney proximal tubule, blood-brain barrier and skeletal muscle. Scientific Reports 2017;7:44517.	R835738 (2016) R835738 (2017) R835738C002 (2016) R835738C002 (2017) R835738C005 (2017)	Full-text from PubMed Abstract from PubMed Associated PubMed link Abstract: Scientific Reports-Corrigendum Exit
Journal Article	Weber EJ, Chapron A, Chapron BD, Voellinger JL, Lidberg KA, Yeung CK, Wang Z, Yamaura Y, Hailey DW, Neumann T, Shen DD, Thummel KE, Muczynski KA, Himmelfarb J, Kelly EJ. Development of a microphysiological model of human kidney proximal tubule function. Kidney International 2016;90(3):627-637.	R835738 (2016) R835738 (2017) R835738C002 (2016) R835738C002 (2017)	Full-text from PubMed Abstract from PubMed Associated PubMed link Abstract: Kidney International-Abstract Exit

Supplemental Keywords:

Kidney, engineered nanomaterials, quantam dots, metals, kidney injury, KIM-I (Kidney Injury Molecule)

Relevant Websites:

The Predictive Toxicology Center (PTC) for Organotypic Cultures Exit

Progress and Final Reports:

Original Abstract

Main Center Abstract and Reports:

R835738 Predictive Toxicology Center for Organotypic Cultures and Assessment of AOPs for Engineered Nanomaterials

Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R835738C001 Airway Epithelium Organotypic Culture as a Platform forAdverseOutcomesPathway Assessment of Engineered Nanomaterials
R835738C002 Organotypic Model of Human Kidney as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
R835738C003 Organotypic Models of Mammalian Liver as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
R835738C004 Organotypic Model of Testis as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
R835738C005 Integrating Liver, Kidney and Testis Nanomaterial Toxicity using the Adverse Outcome Pathway Approach

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.