1 Scientific RepoRts | 7:42296 | DOI: 10.1038/srep42296 www.nature.com/scientificreports 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,† organ interactions resulting from drug, metabolite or xenobiotic transport between organs are key components of human metabolism that impact therapeutic action and toxic side efects. 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 efciency of the drug development process. In this study, fve human MPS models were evaluated for functional coupling, defned 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- specifc processing: terfenadine for pharmacokinetics (PK) and toxicity; trimethylamine (TMA) as a potentially toxic microbiome metabolite; and vitamin D3. We show that the organ-specifc 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 ofer an approach to coupling MPS with distinct media and perfusion requirements. Te goal of in vitro and in vivo toxicity testing is to identify compounds that would predict adverse reactions in humans. Olson et al. 1 found that only 70% of human toxicity was predicted from animal testing. Currently we rely on traditional toxicity testing in animals, a 1930’s methodology that is now challenged due to questionable relevance to human risk, high cost, ethical concerns, and throughput that is too limited for the nearly 80,000 1 University of Pittsburgh, Drug Discovery institute Pittsburgh, PA, USA. 2 Department of computational and Systems Biology, University of Pittsburgh, Baltimore, PA, USA. 3 Departments of Physiology and Medicine, Gi Division, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA. 4 Departments of Molecular Virology and Microbiology and Medicine, Baylor college of Medicine, Houston, tX, USA. 5 Department of Physics and Astronomy, Vanderbilt institute for integrative Biosystems Research and education, Vanderbilt University, nashville, tn, USA. 6 Department of Pharmaceutics, University of Washington, WA, USA. 7 Department of Biomedical engineering, Duke University, Durham, nc, USA. 8 Department of Pharmacy, University of Washington, WA, USA. 9 Kidney Research institute, University of Washington, WA, USA. 10 Department of Medicine, University of Washington, WA, USA. 11 Department of Biomedical engineering, Vanderbilt University, nashville, tn, USA. 12 University of Pittsburgh cancer institute, PA, USA. * these authors contributed equally to this work. † these authors jointly supervised this work. correspondence and requests for materials should be addressed to L.V. (email: Vernetti@pitt.edu) Received: 05 October 2016 Accepted: 20 December 2016 Published: 08 February 2017 opeN