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. 2017 Feb 8:7:42296.
doi: 10.1038/srep42296.

Functional Coupling of Human Microphysiology Systems: Intestine, Liver, Kidney Proximal Tubule, Blood-Brain Barrier and Skeletal Muscle

Lawrence Vernetti  1   2 , Albert Gough  1   2 , Nicholas Baetz  3 , Sarah Blutt  4 , James R Broughman  4 , Jacquelyn A Brown  5 , Jennifer Foulke-Abel  3 , Nesrin Hasan  3 , Julie In  3 , Edward Kelly  6 , Olga Kovbasnjuk  3 , Jonathan Repper  7 , Nina Senutovitch  1 , Janet Stabb  3 , Catherine Yeung  8   9 , Nick C Zachos  3 , Mark Donowitz  3 , Mary Estes  4 , Jonathan Himmelfarb  9   10 , George Truskey  7 , John P Wikswo  5   11 , D Lansing Taylor  1   2   12
Affiliations

Functional Coupling of Human Microphysiology Systems: Intestine, Liver, Kidney Proximal Tubule, Blood-Brain Barrier and Skeletal Muscle

Lawrence Vernetti et al. Sci Rep. .

Erratum in

Abstract

Organ interactions resulting from drug, metabolite or xenobiotic transport between organs are key components of human metabolism that impact therapeutic action and toxic side effects. Preclinical animal testing often fails to predict adverse outcomes arising from sequential, multi-organ metabolism of drugs and xenobiotics. Human microphysiological systems (MPS) can model these interactions and are predicted to dramatically improve the efficiency of the drug development process. In this study, five human MPS models were evaluated for functional coupling, defined as the determination of organ interactions via an in vivo-like sequential, organ-to-organ transfer of media. MPS models representing the major absorption, metabolism and clearance organs (the jejunum, liver and kidney) were evaluated, along with skeletal muscle and neurovascular models. Three compounds were evaluated for organ-specific processing: terfenadine for pharmacokinetics (PK) and toxicity; trimethylamine (TMA) as a potentially toxic microbiome metabolite; and vitamin D3. We show that the organ-specific processing of these compounds was consistent with clinical data, and discovered that trimethylamine-N-oxide (TMAO) crosses the blood-brain barrier. These studies demonstrate the potential of human MPS for multi-organ toxicity and absorption, distribution, metabolism and excretion (ADME), provide guidance for physically coupling MPS, and offer an approach to coupling MPS with distinct media and perfusion requirements.

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Conflict of interest statement

The authors declare no competing financial interests.

Figures

Figure 1
Figure 1. Schematic representations of the four of the organ systems used for functional coupling.
(A) The intestinal module is constructed in transwells from primary jejunum enteroids. Test agents are applied in the apical compartment <1>. The media collected in the basolateral compartment <2> is used to add to the liver. (B) Media from the jejunum intestine basolateral compartment <2> is perfused as a 1:3 jejunum/naïve liver media into the influx port of the SQL-SAL liver model <3>. Efflux media is collected <4> and used to add to two downstream organ models. (C) The vascularized kidney proximal tubule module is a two lumen, dual perfusion system. For the vascular compartment, jejunum/liver-conditioned media <4> is diluted 1:2 or 1:4 with naïve EGM-2 media and then perfused into the influx port <5> to collect effluent from the proximal tubule at <6>. In parallel with perfusion through the vascular compartment, the proximal tubule compartment is perfused with naïve DMEM/F12 PTEC media <6> for effluent collection. (D) The blood-brain barrier with NVU is constructed in a membrane-separated, two-chambered microfluidic device. The brain-derived endothelial vascular compartment is perfused at the influx port <7> with jejunum/liver-conditioned media <4> diluted 1:4 with naïve EGM-2 media. The effluent is collected at the efflux port <7>. In parallel with perfusion through the vascular compartment, the neuronal cell compartment is perfused with naïve EBM-2 media at the influx port <8> for effluent collection at <8>.
Figure 2
Figure 2. Work flow for functional coupling experiments.
Terfenadine exposure is used as an example; TMA and vitamin D3 experiments followed essentially the same workflow. The test compound is initially added to the apical gut media and samples are collected from the apical and basolateral media for MS analysis. Basolateral media samples are sent to UPitt where they are mixed with liver media for exposure in the liver module. Effluent samples are taken for MS analysis and sent to UWash and Vanderbilt where they are mixed with kidney and NVU media, respectively. Samples are taken of the effluent from the kidney proximal tubule module and from the vascular and brain sides of the NVU for MS analysis.
Figure 3
Figure 3. Functional analysis of terfenadine transport and metabolism in human organs.
Terfenadine taken orally is absorbed and metabolized to fexofenadine in the intestine. Counter-transport carries fexofenadine back to the apical side of the intestinal wall. Remaining terfenadine is metabolized to fexofenadine in the liver. Fexofenadine is not able to cross the blood-brain barrier and is excreted by the kidney.
Figure 4
Figure 4. Effect of functional integration on terfenadine toxicity.
Pre-conditioned terfenadine in the static liver model results in decreased effect on skeletal myobundles. Results are for one donor, N = 3 and presented as Mean ± S.D. *p < 0.05, #p < 0.01.
Figure 5
Figure 5. Functional analysis of TMA transport and metabolism in human organs.
TMA produced in the intestine by the endogenous microbiome is taken up and transported to the liver, where it is metabolized to TMAO. Kidney disease can lead to increased plasma concentrations of TMAO, with significant medical side effects.
Figure 6
Figure 6. Functional analysis of Vitamin D3 transport and metabolism in human organs.
Absorption and activation of Vitamin D3 requires transport and metabolism in multiple organs. Sequential metabolism in the liver and the kidney leads to a series of active metabolites. Vitamin D3 and its metabolites are hydrophobic and therefore can easily be lost in MPS systems due to binding to materials like PDMS. Use of a carrier protein is important in MPS devices for physiological relevance.
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