Grantee Research Project Results

2016 Progress Report: Systems Engineering & Analysis for Organotypic Culture Models

EPA Grant Number: R835736C005
Subproject: this is subproject number 005 , established and managed by the Center Director under grant R835736
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: Mickey Leland National Urban Air Toxics Research Center (NUATRC)
Center Director: Beskid, Craig
Title: Systems Engineering & Analysis for Organotypic Culture Models
Investigators: Wikswo, John , Cliffel, David , McLean, John , Shotwell, Matt
Institution: Vanderbilt University
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:

Project 5 is designed to ensure physiologically realistic function of each of the mammary, limb, fetal membrane, and liver organotypic culture models (OCMs), operating separately and in combination, and will do so by coordinating the refinement of Integrated Organ Microfluidics (IOM) modules that will provide a common platform architecture.

Progress Summary:

Project 5 continues to deliver standalone pumps, Perfusion Controller (IOM-PC, v1.0), and MicroClinical Analyzer (µCA, v1.0) hardware integration modules as requested by organ developers. We also deliver limb segmentation (OCM 3.0) prototypes to Project 2 as described in Milestone P2.1. We are looking into materials to replace polydimethylsiloxane (PDMS) in pump and valve fabrication.

Milestone 2.1 (carry-over): Develop offline metabolic secretion baselines for each OCM (IM-MS, μCA). We have received and analyzed effluent for metabolic baseline readouts via ion mobility-mass spectrometry (IM-MS) from Project 4 (liver). Results were shown in our last annual SAC report. The liver team also has shipped samples (November 2016) for baseline analysis using the MicroClinical Analyzer (µCA). We have not received samples from Projects 1, 3 for IM-MS or Projects 1, 2, 3 for µCA baseline analysis, but are ready to do so at any time. Project 2 elected not to conduct IM-MS analysis as it is not a milestone requirement for their OCM.

Milestone 2.4: Version 2.0 IOM-μCA integration. We currently provide and support µCA v1.0 modules, with 25 µL sensor volumes, as required by project investigators. We have developed sensors for glutamate, and are developing ones for cholesterol and ammonium. We now recognize that we should prevent bubbles from entering the sensor, and are learning how to avoid the effects of sensor interferents such as glycine and ascorbic acid. Advances in screen-printed electrode (SPE) fabrication and design allow us to reduce the sensor volume of our µCA v2.0 to 6 µL, and new custom, eight-analyte SPEs have been ordered from Pine instruments and we have designed the microfluidics for the sensor housing.

Extensive testing of pumps and valves produced with parametric variations of masters has allowed us to finalize both RPPM and RPV designs for the µCAs and organ-chip perfusion controllers. We have refined our processes to produce Rotary Planar Peristaltic Micropumps (RPPMs) that span a range of flow rate from 0.5 to 800 µL/min, and circular-segment 5-port and 24-port rotary planar valves (RPVs). These will be used in IOM-μCA V2.0 and 3.0.

Milestone 2.5: Version 2.0 IOM-Microformulator integration (µF). We have developed 26-position RPVs. We also have designs for an integrated recirculation valve and an organ interconnection valve that will be tested as soon as the triple NEMA-8 SmartMotor cartridges are fully operational. Most important, we have recognized that four of the 24-channel MicroFormulators that can be created with a 5-port RPV, an RPPM, and a 24-port RPV can be used to individually address each well and insert in a 24-transwell plate, and enable automatic measurement of the TransEndothelial Electrical Resistance (TEER) of each insert membrane.

Milestone 3.1: Software will enable full automation of IOMs. We continue to test and refine our Automated Multi-Pump Experiment Running Environment (AMPERE) software as we receive user feedback. We have developed an AMPERE wrapper that controls 6- and 24-motor, 24- and 96-channel µFs to provide timed toxin exposure, including synthesis of pharmacokinetic (PK) exposure profiles. AMPERE will control TEER electronics for the fetal membrane chip.

Milestone 3.2: Integrate wireless capability to support many IOMs. With funding from NIH/NCATS and AstraZeneca, we are nearing completion of our first prototype Triple NEMA-8 Wireless SmartMotor Cartridge. As soon as this prototype is complete, we will begin production of cartridges that will be used during 2017, with EPA funding, to create all Perfusion Controllers, MicroClinical Analyzers, and MicroFormulators that will be used by Projects 1-4.

Milestone 3.3: Develop prediction models for organ health and chemical toxicity based on baseline metabolic secretions. We have recommended a uniform protocol for determining AC50s and maximal responses based on nonlinear least squares regression methods. We have discussed potential methodologies for clustering metrics and for constructing meaningful prediction models. As training data from OCM chemical exposure experiments become available, we will ramp up efforts under this milestone. In addition, we are collaborating with researchers at EPA’s National Center for Computational Toxicology on cell-agent-based models of how chemicals disrupt particular events during embryogenesis.

Future Activities:

Continued refinement of device integration and molding to reduce cost.

Journal Articles on this Report : 2 Displayed | Download in RIS Format

Publications Views
Other subproject views:	All 38 publications	8 publications in selected types	All 8 journal articles
Other center views:	All 169 publications	57 publications in selected types	All 56 journal articles

Publications
Type	Citation	Sub Project	Document Sources
Journal Article	Leung MC, Hutson MS, Seifert AW, Spencer RM, Knudsen TB. Computational modeling and simulation of genital tubercle development. Reproductive Toxicology 2016;64:151-161.	R835736 (2015) R835736 (2016) R835736C005 (2016)	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 (15 pp.).	R835736 (2015) R835736 (2016) R835736C004 (2016) R835736C005 (2016) R835738 (2016) R835738C003 (2017) R835738C005 (2017)	Full-text from PubMed Abstract from PubMed Associated PubMed link Full-text: Nature-Full Text PDF Exit Abstract: Nature-Abstract Exit Other: Scientific Reports-Full Text PDF Exit

Supplemental Keywords:

Microfluidic pumps and valves, integrated organ microfluidics.

Relevant Websites:

VIIBRE, the Vanderbilt Institute for Integrative Biosystems Research and Education Exit

Vanderbilt-Pittsburgh Resource for Organotypic Models for Predictive Toxicology (VPROMPT) Exit

Progress and Final Reports:

Original Abstract

Main Center Abstract and Reports:

R835736 Mickey Leland National Urban Air Toxics Research Center (NUATRC)

Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R835736C001 Mammosphere Bioreactor For Life-Stage Specific Toxicology
R835736C002 Organotypic Culture Model to Analyze DevelopmentalLimbMalformationsResulting from Toxicant/Teratogen Exposure
R835736C003 Validating a fetal membrane on a chip model for characterizing reproductive toxicant exposure risks
R835736C004 Organotypic Liver Model for Predictive Human Toxicology and Metabolism
R835736C005 Systems Engineering & Analysis for Organotypic Culture Models

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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.