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

Lawrence Vernetti,D Lansing Taylor,Janet Stabb,James R Broughman,Jonathan Himmelfarb,John P Wikswo,Jonathan Repper,Edward J Kelly ,Sarah E Blutt ,Nesrin M Hasan ,Nicholas W Baetz ,Catherine K Yeung ,Jennifer Foulke‐Abel ,Mary K Estes ,George A Truskey ,Mark Donowitz,Olga Kovbasnjuk,Nick C Zachos,Nina Senutovitch,Julie In,Jacquelyn A Brown,Albert Gough

doi:10.1038/srep42296

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.

Highlights

MethodsA summary of methods used for the preparation and functional coupling in the four-organ and two-organ experiments follows
Skeletal muscle myobundle models terfenadine toxicity was reduced by metabolism in the liver, consistent with clinical toxicity
These results were consistent with the known in vivo metabolism and transport of these compounds (Table 2), with a new finding that TMAO crosses the BBB

Summary

Methods

A summary of methods used for the preparation and functional coupling in the four-organ and two-organ experiments follows. More detailed methods and dimensions of the individual devices are provided in the Supplemental Material and Supplemental Table S7. The human liver, kidney, NVU and skeletal muscle organ constructs have established long term viability for at least 28 days in culture[52,53,54]. Mass spectrometry was used to measure the concentration of compound and metabolites in the media following exposure in each MPS organ module. All Vitamin D3 samples were analyzed at U Washington using an established method[52,56]. All TMA, TMAO samples were analyzed at U Washington using a novel LC MS/MS method (unpublished). Terfenadine/fexofenadine mass spectrometry analyses were conducted at UPitt, U Washington, Duke and Vanderbilt using previously published methods[57,58]

Results

Discussion

Conclusion