{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,6]],"date-time":"2026-11-06T04:25:10Z","timestamp":1793939110899,"version":"4.8.0"},"reference-count":75,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,11,23]],"date-time":"2021-11-23T00:00:00Z","timestamp":1637625600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,11,23]],"date-time":"2021-11-23T00:00:00Z","timestamp":1637625600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Non-alcoholic steatohepatitis (NASH) is a progressive and severe liver disease, characterized by lipid accumulation, inflammation, and downstream fibrosis. Despite its increasing prevalence, there is no approved treatment yet available for patients. This has been at least partially due to the lack of predictive preclinical models for studying this complex disease. Here, we present a 3D in vitro microtissue model that uses spheroidal, scaffold free co-culture of primary human hepatocytes, Kupffer cells, liver endothelial cells and hepatic stellate cells. Upon exposure to defined and clinically relevant lipotoxic and inflammatory stimuli, these microtissues develop key pathophysiological features of NASH within 10\u00a0days, including an increase of intracellular triglyceride content and lipids, and release of pro-inflammatory cytokines. Furthermore, fibrosis was evident through release of procollagen type I, and increased deposition of extracellular collagen fibers. Whole transcriptome analysis revealed changes in the regulation of pathways associated with NASH, such as lipid metabolism, inflammation and collagen processing. Importantly, treatment with anti-NASH drug candidates (Selonsertib and Firsocostat) decreased the measured specific disease parameter, in accordance with clinical observations. These drug treatments also significantly changed the gene expression patterns of the microtissues, thus providing mechanisms of action and revealing therapeutic potential. In summary, this human NASH model represents a promising drug discovery tool for understanding the underlying complex mechanisms in NASH, evaluating efficacy of anti-NASH drug candidates and identifying new approaches for therapeutic interventions.<\/jats:p>","DOI":"10.1038\/s41598-021-01951-7","type":"journal-article","created":{"date-parts":[[2021,11,23]],"date-time":"2021-11-23T06:02:20Z","timestamp":1637647340000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":55,"title":["A 3D primary human cell-based in vitro model of non-alcoholic steatohepatitis for efficacy testing of clinical drug candidates"],"prefix":"10.1038","volume":"11","author":[{"given":"Simon","family":"Str\u00f6bel","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Radina","family":"Kostadinova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katia","family":"Fiaschetti-Egli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jana","family":"Rupp","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manuela","family":"Bieri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Agnieszka","family":"Pawlowska","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donna","family":"Busler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Hofstetter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katarzyna","family":"Sanchez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sue","family":"Grepper","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Thoma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,11,23]]},"reference":[{"issue":"7","key":"1951_CR1","doi-asserted-by":"publisher","first-page":"908","DOI":"10.1038\/s41591-018-0104-9","volume":"24","author":"SL Friedman","year":"2018","unstructured":"Friedman, S. L. et al. Mechanisms of NAFLD development and therapeutic strategies. Nat. Med. 24(7), 908\u2013922 (2018).","journal-title":"Nat. Med."},{"issue":"7","key":"1951_CR2","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1038\/nrgastro.2017.38","volume":"14","author":"T Tsuchida","year":"2017","unstructured":"Tsuchida, T. & Friedman, S. L. Mechanisms of hepatic stellate cell activation. Nat. Rev. Gastroenterol. Hepatol. 14(7), 397\u2013411 (2017).","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"issue":"8","key":"1951_CR3","doi-asserted-by":"publisher","first-page":"1038","DOI":"10.1016\/j.metabol.2015.12.012","volume":"65","author":"E Buzzetti","year":"2016","unstructured":"Buzzetti, E., Pinzani, M. & Tsochatzis, E. A. The multiple-hit pathogenesis of non-alcoholic fatty liver disease (NAFLD). Metabolism 65(8), 1038\u20131048 (2016).","journal-title":"Metabolism"},{"issue":"2","key":"1951_CR4","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1016\/j.cld.2015.10.001","volume":"20","author":"M Sayiner","year":"2016","unstructured":"Sayiner, M. et al. Epidemiology of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis in the United States and the Rest of the World. Clin. Liver Dis. 20(2), 205\u2013214 (2016).","journal-title":"Clin. Liver Dis."},{"issue":"4","key":"1951_CR5","doi-asserted-by":"publisher","first-page":"390","DOI":"10.1016\/j.jceh.2018.04.001","volume":"8","author":"AJ Kovalic","year":"2018","unstructured":"Kovalic, A. J. et al. Genetic and epigenetic culprits in the pathogenesis of nonalcoholic fatty liver disease. J. Clin. Exp. Hepatol. 8(4), 390\u2013402 (2018).","journal-title":"J. Clin. Exp. Hepatol."},{"issue":"2","key":"1951_CR6","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1080\/13543784.2020.1668374","volume":"29","author":"N Alkhouri","year":"2020","unstructured":"Alkhouri, N. et al. GS-0976 (Firsocostat): An investigational liver-directed acetyl-CoA carboxylase (ACC) inhibitor for the treatment of non-alcoholic steatohepatitis (NASH). Expert Opin. Investig. Drugs 29(2), 135\u2013141 (2020).","journal-title":"Expert Opin. Investig. Drugs"},{"issue":"5","key":"1951_CR7","doi-asserted-by":"publisher","first-page":"2241","DOI":"10.1002\/hep.30333","volume":"69","author":"G Farrell","year":"2019","unstructured":"Farrell, G. et al. Mouse models of nonalcoholic steatohepatitis: Toward optimization of their relevance to human nonalcoholic steatohepatitis. Hepatology 69(5), 2241\u20132257 (2019).","journal-title":"Hepatology"},{"issue":"5","key":"1951_CR8","doi-asserted-by":"publisher","first-page":"943","DOI":"10.1016\/j.bbadis.2018.06.023","volume":"1865","author":"D Jahn","year":"2019","unstructured":"Jahn, D. et al. Animal models of NAFLD from a hepatologist\u2019s point of view. Biochim. Biophys. Acta Mol. Basis Dis. 1865(5), 943\u2013953 (2019).","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"1951_CR9","doi-asserted-by":"crossref","unstructured":"Even, P.C., et al. Editorial: Are rodent models fit for investigation of human obesity and related diseases? Front. Nutr. 4, 58 (2017).","DOI":"10.3389\/fnut.2017.00058"},{"issue":"5","key":"1951_CR10","doi-asserted-by":"publisher","first-page":"1701","DOI":"10.1002\/hep.31517","volume":"73","author":"Y Ma","year":"2021","unstructured":"Ma, Y. et al. 17-Beta hydroxysteroid dehydrogenase 13 deficiency does not protect mice from obesogenic diet injury. Hepatology 73(5), 1701\u20131716 (2021).","journal-title":"Hepatology"},{"key":"1951_CR11","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1186\/s12986-020-00462-y","volume":"17","author":"X Xin","year":"2020","unstructured":"Xin, X. et al. High-trans fatty acid and high-sugar diets can cause mice with non-alcoholic steatohepatitis with liver fibrosis and potential pathogenesis. Nutr. Metab. (Lond.) 17, 40 (2020).","journal-title":"Nutr. Metab. (Lond.)"},{"issue":"5","key":"1951_CR12","doi-asserted-by":"publisher","first-page":"1836","DOI":"10.1002\/hep.24001","volume":"52","author":"H Tilg","year":"2010","unstructured":"Tilg, H. & Moschen, A. R. Evolution of inflammation in nonalcoholic fatty liver disease: the multiple parallel hits hypothesis. Hepatology 52(5), 1836\u20131846 (2010).","journal-title":"Hepatology"},{"issue":"14","key":"1951_CR13","doi-asserted-by":"publisher","first-page":"e2007425","DOI":"10.1002\/smll.202007425","volume":"17","author":"HJ Cho","year":"2021","unstructured":"Cho, H. J. et al. Bioengineered multicellular liver microtissues for modeling advanced hepatic fibrosis driven through non-alcoholic fatty liver disease. Small 17(14), e2007425 (2021).","journal-title":"Small"},{"key":"1951_CR14","doi-asserted-by":"publisher","first-page":"257","DOI":"10.1016\/j.phrs.2018.06.029","volume":"134","author":"J Boeckmans","year":"2018","unstructured":"Boeckmans, J. et al. Human-based systems: Mechanistic NASH modelling just around the corner?. Pharmacol. Res. 134, 257\u2013267 (2018).","journal-title":"Pharmacol. Res."},{"key":"1951_CR15","doi-asserted-by":"publisher","first-page":"357","DOI":"10.3389\/fonc.2018.00357","volume":"8","author":"I Fabregat","year":"2018","unstructured":"Fabregat, I. & Caballero-D\u00edaz, D. Transforming growth factor-\u03b2-induced cell plasticity in liver fibrosis and hepatocarcinogenesis. Front. Oncol. 8, 357 (2018).","journal-title":"Front. Oncol."},{"issue":"8","key":"1951_CR16","doi-asserted-by":"publisher","first-page":"457","DOI":"10.1038\/s41575-020-0304-x","volume":"17","author":"P Ramachandran","year":"2020","unstructured":"Ramachandran, P. et al. Single-cell technologies in hepatology: New insights into liver biology and disease pathogenesis. Nat. Rev. Gastroenterol. Hepatol. 17(8), 457\u2013472 (2020).","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"issue":"3","key":"1951_CR17","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1038\/s41575-018-0082-x","volume":"16","author":"K Kazankov","year":"2019","unstructured":"Kazankov, K. et al. The role of macrophages in nonalcoholic fatty liver disease and nonalcoholic steatohepatitis. Nat. Rev. Gastroenterol. Hepatol. 16(3), 145\u2013159 (2019).","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"1951_CR18","doi-asserted-by":"crossref","unstructured":"Dewidar, B., et al. TGF-\u03b2 in hepatic stellate cell activation and liver fibrogenesis-updated 2019. Cells. 8(11), 1419 (2019).","DOI":"10.3390\/cells8111419"},{"issue":"4","key":"1951_CR19","doi-asserted-by":"publisher","first-page":"326","DOI":"10.1124\/dmd.117.078675","volume":"46","author":"N Tanner","year":"2018","unstructured":"Tanner, N. et al. Regulation of drug metabolism by the interplay of inflammatory signaling, steatosis, and xeno-sensing receptors in HepaRG cells. Drug Metab. Dispos. 46(4), 326\u2013335 (2018).","journal-title":"Drug Metab. Dispos."},{"key":"1951_CR20","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1016\/j.lfs.2018.04.022","volume":"203","author":"X Chen","year":"2018","unstructured":"Chen, X. et al. Oleic acid protects saturated fatty acid mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis. Life Sci. 203, 291\u2013304 (2018).","journal-title":"Life Sci."},{"issue":"1","key":"1951_CR21","doi-asserted-by":"publisher","first-page":"11585","DOI":"10.1038\/s41598-019-47737-w","volume":"9","author":"S Prill","year":"2019","unstructured":"Prill, S. et al. The TM6SF2 E167K genetic variant induces lipid biosynthesis and reduces apolipoprotein B secretion in human hepatic 3D spheroids. Sci. Rep. 9(1), 11585 (2019).","journal-title":"Sci. Rep."},{"issue":"20","key":"1951_CR22","doi-asserted-by":"publisher","first-page":"e90954","DOI":"10.1172\/jci.insight.90954","volume":"1","author":"RE Feaver","year":"2016","unstructured":"Feaver, R. E. et al. Development of an in vitro human liver system for interrogating nonalcoholic steatohepatitis. JCI Insight. 1(20), e90954 (2016).","journal-title":"JCI Insight."},{"issue":"3","key":"1951_CR23","first-page":"1531","volume":"11","author":"S Mukherjee","year":"2019","unstructured":"Mukherjee, S. et al. Development and validation of an in vitro 3D model of NASH with severe fibrotic phenotype. Am. J. Transl. Res. 11(3), 1531\u20131540 (2019).","journal-title":"Am. J. Transl. Res."},{"issue":"5","key":"1951_CR24","doi-asserted-by":"publisher","first-page":"1063","DOI":"10.1016\/j.jhep.2018.01.019","volume":"68","author":"T Jensen","year":"2018","unstructured":"Jensen, T. et al. Fructose and sugar: A major mediator of non-alcoholic fatty liver disease. J. Hepatol. 68(5), 1063\u20131075 (2018).","journal-title":"J. Hepatol."},{"issue":"2","key":"1951_CR25","doi-asserted-by":"publisher","first-page":"470","DOI":"10.1002\/hep.31056","volume":"72","author":"G Carpino","year":"2020","unstructured":"Carpino, G. et al. Increased liver localization of lipopolysaccharides in human and experimental NAFLD. Hepatology 72(2), 470\u2013485 (2020).","journal-title":"Hepatology"},{"issue":"9\u201310","key":"1951_CR26","doi-asserted-by":"publisher","first-page":"1172","DOI":"10.1002\/biot.200700228","volume":"3","author":"RZ Lin","year":"2008","unstructured":"Lin, R. Z. & Chang, H. Y. Recent advances in three-dimensional multicellular spheroid culture for biomedical research. Biotechnol. J. 3(9\u201310), 1172\u20131184 (2008).","journal-title":"Biotechnol. J."},{"issue":"4","key":"1951_CR27","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1016\/j.tibtech.2004.02.002","volume":"22","author":"JM Kelm","year":"2004","unstructured":"Kelm, J. M. & Fussenegger, M. Microscale tissue engineering using gravity-enforced cell assembly. Trends Biotechnol. 22(4), 195\u2013202 (2004).","journal-title":"Trends Biotechnol."},{"issue":"1","key":"1951_CR28","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1007\/s00204-012-0968-2","volume":"87","author":"S Messner","year":"2013","unstructured":"Messner, S. et al. Multi-cell type human liver microtissues for hepatotoxicity testing. Arch Toxicol. 87(1), 209\u2013213 (2013).","journal-title":"Arch Toxicol."},{"issue":"4","key":"1951_CR29","doi-asserted-by":"publisher","first-page":"896","DOI":"10.1016\/j.jhep.2020.04.037","volume":"73","author":"J Bates","year":"2020","unstructured":"Bates, J. et al. Acetyl-CoA carboxylase inhibition disrupts metabolic reprogramming during hepatic stellate cell activation. J. Hepatol. 73(4), 896\u2013905 (2020).","journal-title":"J. Hepatol."},{"issue":"4","key":"1951_CR30","doi-asserted-by":"publisher","first-page":"829","DOI":"10.1016\/j.jcmgh.2020.06.001","volume":"10","author":"TT Ross","year":"2020","unstructured":"Ross, T. T. et al. Acetyl-CoA carboxylase inhibition improves multiple dimensions of NASH pathogenesis in model systems. Cell Mol. Gastroenterol. Hepatol. 10(4), 829\u2013851 (2020).","journal-title":"Cell Mol. Gastroenterol. Hepatol."},{"issue":"5","key":"1951_CR31","doi-asserted-by":"publisher","first-page":"1463","DOI":"10.1053\/j.gastro.2018.07.027","volume":"155","author":"R Loomba","year":"2018","unstructured":"Loomba, R. et al. GS-0976 reduces hepatic steatosis and fibrosis markers in patients with nonalcoholic fatty liver disease. Gastroenterology 155(5), 1463-1473.e6 (2018).","journal-title":"Gastroenterology"},{"issue":"12","key":"1951_CR32","doi-asserted-by":"publisher","first-page":"1983","DOI":"10.1016\/j.cgh.2018.04.042","volume":"16","author":"EJ Lawitz","year":"2018","unstructured":"Lawitz, E. J. et al. Acetyl-CoA carboxylase inhibitor GS-0976 for 12 weeks reduces hepatic de novo lipogenesis and steatosis in patients with nonalcoholic steatohepatitis. Clin. Gastroenterol. Hepatol. 16(12), 1983-1991.e3 (2018).","journal-title":"Clin. Gastroenterol. Hepatol."},{"issue":"6","key":"1951_CR33","doi-asserted-by":"publisher","first-page":"527","DOI":"10.4062\/biomolther.2020.016","volume":"28","author":"YC Yoon","year":"2020","unstructured":"Yoon, Y. C. et al. Selonsertib inhibits liver fibrosis via downregulation of ASK1\/ MAPK pathway of hepatic stellate cells. Biomol. Ther. (Seoul) 28(6), 527\u2013536 (2020).","journal-title":"Biomol. Ther. (Seoul)"},{"issue":"4","key":"1951_CR34","doi-asserted-by":"publisher","first-page":"1377","DOI":"10.1002\/hep.30651","volume":"70","author":"HL Paish","year":"2019","unstructured":"Paish, H. L. et al. A bioreactor technology for modeling fibrosis in human and rodent precision-cut liver slices. Hepatology 70(4), 1377\u20131391 (2019).","journal-title":"Hepatology"},{"issue":"2","key":"1951_CR35","doi-asserted-by":"publisher","first-page":"362","DOI":"10.1016\/j.bbrc.2020.07.141","volume":"530","author":"M Sakai","year":"2020","unstructured":"Sakai, M. et al. GPR91 antagonist and TGF-\u03b2 inhibitor suppressed collagen production of high glucose and succinate induced HSC activation. Biochem. Biophys. Res. Commun. 530(2), 362\u2013366 (2020).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"1951_CR36","volume-title":"Romeis\u2014Mikroskopische Technik","author":"M Mulisch","year":"2010","unstructured":"Mulisch, M. & Welsch, U. Romeis\u2014Mikroskopische Technik (Spektrum Akademischer Verlag, 2010)."},{"key":"1951_CR37","volume-title":"Histological and Histochemical Methods: Theory and Practice","author":"JA Kiernan","year":"1999","unstructured":"Kiernan, J. A. Histological and Histochemical Methods: Theory and Practice 3rd edn. (Butterworth-Heinemann, 1999).","edition":"3"},{"key":"1951_CR38","unstructured":"Biospyder website. https:\/\/www.biospyder.com\/"},{"key":"1951_CR39","volume-title":"R: A Language and Environment for Statistical Computing","author":"Team RC","year":"2021","unstructured":"Team RC. R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, 2021)."},{"issue":"12","key":"1951_CR40","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","volume":"15","author":"MI Love","year":"2014","unstructured":"Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 15(12), 550 (2014).","journal-title":"Genome Biol."},{"key":"1951_CR41","unstructured":"Leek, J.T., et al., sva: Surrogate Variable Analysis,\" (2020). R package version 3.36.0."},{"issue":"43","key":"1951_CR42","doi-asserted-by":"publisher","first-page":"15545","DOI":"10.1073\/pnas.0506580102","volume":"102","author":"A Subramanian","year":"2005","unstructured":"Subramanian, A. et al. Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl. Acad. Sci. USA 102(43), 15545\u201315550 (2005).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"issue":"5","key":"1951_CR43","doi-asserted-by":"publisher","first-page":"284","DOI":"10.1089\/omi.2011.0118","volume":"16","author":"G Yu","year":"2012","unstructured":"Yu, G. et al. clusterProfiler: An R package for comparing biological themes among gene clusters. OMICS 16(5), 284\u2013287 (2012).","journal-title":"OMICS"},{"issue":"12","key":"1951_CR44","doi-asserted-by":"publisher","first-page":"1739","DOI":"10.1093\/bioinformatics\/btr260","volume":"27","author":"A Liberzon","year":"2011","unstructured":"Liberzon, A. et al. Molecular signatures database (MSigDB) 3.0. Bioinformatics 27(12), 1739\u20131340 (2011).","journal-title":"Bioinformatics"},{"issue":"1","key":"1951_CR45","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1093\/nar\/28.1.27","volume":"28","author":"M Kanehisa","year":"2000","unstructured":"Kanehisa, M. & Goto, S. KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Res. 28(1), 27\u201330 (2000).","journal-title":"Nucleic Acids Res."},{"issue":"Database issue","key":"1951_CR46","doi-asserted-by":"publisher","first-page":"D674","DOI":"10.1093\/nar\/gkn653","volume":"37","author":"CF Schaefer","year":"2009","unstructured":"Schaefer, C. F. et al. PID: The pathway interaction database. Nucleic Acids Res 37(Database issue), D674\u2013D679 (2009).","journal-title":"Nucleic Acids Res"},{"issue":"D1","key":"1951_CR47","first-page":"D498","volume":"48","author":"B Jassal","year":"2020","unstructured":"Jassal, B. et al. The reactome pathway knowledgebase. Nucleic Acids Res. 48(D1), D498-d503 (2020).","journal-title":"Nucleic Acids Res."},{"issue":"D1","key":"1951_CR48","doi-asserted-by":"publisher","first-page":"D613","DOI":"10.1093\/nar\/gkaa1024","volume":"49","author":"M Martens","year":"2020","unstructured":"Martens, M. et al. WikiPathways: Connecting communities. Nucleic Acids Res. 49(D1), D613\u2013D621 (2020).","journal-title":"Nucleic Acids Res."},{"issue":"1","key":"1951_CR49","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1111\/j.2517-6161.1995.tb02031.x","volume":"57","author":"Y Benjamini","year":"1995","unstructured":"Benjamini, Y. & Hochberg, Y. Controlling the false discovery rate: A practical and powerful approach to multiple testing. J. R. Stat. Soc. Series Methodol. 57(1), 289\u2013300 (1995).","journal-title":"J. R. Stat. Soc. Series Methodol."},{"issue":"1","key":"1951_CR50","first-page":"100","volume":"6","author":"J Grunhut","year":"2018","unstructured":"Grunhut, J. et al. Macrophages in nonalcoholic steatohepatitis: Friend or foe?. Eur. Med. J. Hepatol. 6(1), 100\u2013109 (2018).","journal-title":"Eur. Med. J. Hepatol."},{"issue":"8","key":"1951_CR51","doi-asserted-by":"publisher","first-page":"727","DOI":"10.3748\/wjg.v18.i8.727","volume":"18","author":"V Braunersreuther","year":"2012","unstructured":"Braunersreuther, V. et al. Role of cytokines and chemokines in non-alcoholic fatty liver disease. World J. Gastroenterol. 18(8), 727\u2013735 (2012).","journal-title":"World J. Gastroenterol."},{"key":"1951_CR52","doi-asserted-by":"publisher","first-page":"395","DOI":"10.1007\/978-1-4939-7113-8_26","volume":"1627","author":"L Ritti\u00e9","year":"2017","unstructured":"Ritti\u00e9, L. Method for picrosirius red-polarization detection of collagen fibers in tissue sections. Methods Mol. Biol. 1627, 395\u2013407 (2017).","journal-title":"Methods Mol. Biol."},{"issue":"10","key":"1951_CR53","doi-asserted-by":"publisher","first-page":"2087","DOI":"10.1124\/dmd.109.027466","volume":"37","author":"CD Fisher","year":"2009","unstructured":"Fisher, C. D. et al. Hepatic cytochrome P450 enzyme alterations in humans with progressive stages of nonalcoholic fatty liver disease. Drug Metab. Dispos. 37(10), 2087\u20132094 (2009).","journal-title":"Drug Metab. Dispos."},{"issue":"5","key":"1951_CR54","doi-asserted-by":"publisher","first-page":"2418","DOI":"10.1128\/IAI.68.5.2418-2423.2000","volume":"68","author":"K Imai","year":"2000","unstructured":"Imai, K., Takeshita, A. & Hanazawa, S. Transforming growth factor-beta inhibits lipopolysaccharide-stimulated expression of inflammatory cytokines in mouse macrophages through downregulation of activation protein 1 and CD14 receptor expression. Infect. Immunol. 68(5), 2418\u20132423 (2000).","journal-title":"Infect. Immunol."},{"issue":"4","key":"1951_CR55","first-page":"359","volume":"8","author":"M Duriez","year":"2020","unstructured":"Duriez, M. et al. A 3D human liver model of nonalcoholic steatohepatitis. J. Clin. Transl. Hepatol. 8(4), 359\u2013370 (2020).","journal-title":"J. Clin. Transl. Hepatol."},{"key":"1951_CR56","doi-asserted-by":"crossref","unstructured":"Van Herck, M.A., L. Vonghia, & S.M. Francque. Animal models of nonalcoholic fatty liver disease-a starter's guide. Nutrients. 9(10), 1072 (2017).","DOI":"10.3390\/nu9101072"},{"key":"1951_CR57","doi-asserted-by":"publisher","first-page":"3861","DOI":"10.2147\/DMSO.S249949","volume":"13","author":"MA Hegazy","year":"2020","unstructured":"Hegazy, M. A. et al. Serum LPS and CD163 biomarkers confirming the role of gut dysbiosis in overweight patients with NASH. Diabetes Metab. Syndr. Obes. 13, 3861\u20133872 (2020).","journal-title":"Diabetes Metab. Syndr. Obes."},{"issue":"1","key":"1951_CR58","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/15476278.2018.1423931","volume":"14","author":"Y Kim","year":"2018","unstructured":"Kim, Y. et al. Prolongation of liver-specific function for primary hepatocytes maintenance in 3D printed architectures. Organogenesis 14(1), 1\u201312 (2018).","journal-title":"Organogenesis"},{"key":"1951_CR59","doi-asserted-by":"crossref","unstructured":"Pingitore, P., et al. Human multilineage 3D spheroids as a model of liver steatosis and fibrosis. Int. J. Mol. Sci. 20(7), 1629 (2019).","DOI":"10.3390\/ijms20071629"},{"issue":"1","key":"1951_CR60","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1016\/j.vascn.2010.04.013","volume":"63","author":"M L\u00fcbberstedt","year":"2011","unstructured":"L\u00fcbberstedt, M. et al. HepaRG human hepatic cell line utility as a surrogate for primary human hepatocytes in drug metabolism assessment in vitro. J. Pharmacol. Toxicol. Methods 63(1), 59\u201368 (2011).","journal-title":"J. Pharmacol. Toxicol. Methods"},{"issue":"6","key":"1951_CR61","doi-asserted-by":"publisher","first-page":"e0179995","DOI":"10.1371\/journal.pone.0179995","volume":"12","author":"V Prestigiacomo","year":"2017","unstructured":"Prestigiacomo, V. et al. Pro-fibrotic compounds induce stellate cell activation, ECM-remodelling and Nrf2 activation in a human 3D-multicellular model of liver fibrosis. PLoS ONE 12(6), e0179995 (2017).","journal-title":"PLoS ONE"},{"issue":"1","key":"1951_CR62","doi-asserted-by":"publisher","first-page":"14297","DOI":"10.1038\/s41598-018-32722-6","volume":"8","author":"M Kozyra","year":"2018","unstructured":"Kozyra, M. et al. Human hepatic 3D spheroids as a model for steatosis and insulin resistance. Sci. Rep. 8(1), 14297 (2018).","journal-title":"Sci. Rep."},{"key":"1951_CR63","doi-asserted-by":"crossref","unstructured":"Hurrell, T., et al. Human liver spheroids as a model to study aetiology and treatment of hepatic fibrosis. Cells. 9(4), 964 (2020).","DOI":"10.3390\/cells9040964"},{"issue":"1","key":"1951_CR64","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1002\/hep4.1450","volume":"4","author":"T Kostrzewski","year":"2020","unstructured":"Kostrzewski, T. et al. A microphysiological system for studying nonalcoholic steatohepatitis. Hepatol. Commun. 4(1), 77\u201391 (2020).","journal-title":"Hepatol. Commun."},{"issue":"2","key":"1951_CR65","doi-asserted-by":"publisher","first-page":"204","DOI":"10.3748\/wjg.v23.i2.204","volume":"23","author":"TC TomaszKostrzewski","year":"2017","unstructured":"TomaszKostrzewski, T. C. et al. Three-dimensional perfused human in vitro model of nonalcoholic fatty liver disease. World J. Gastroenterol. 23(2), 204\u2013215 (2017).","journal-title":"World J. Gastroenterol."},{"issue":"8","key":"1951_CR66","doi-asserted-by":"publisher","first-page":"2849","DOI":"10.1007\/s00204-017-2002-1","volume":"91","author":"WR Proctor","year":"2017","unstructured":"Proctor, W. R. et al. Utility of spherical human liver microtissues for prediction of clinical drug-induced liver injury. Arch Toxicol. 91(8), 2849\u20132863 (2017).","journal-title":"Arch Toxicol."},{"issue":"11","key":"1951_CR67","doi-asserted-by":"publisher","first-page":"1360","DOI":"10.1016\/j.molmet.2017.09.001","volume":"6","author":"H Jouihan","year":"2017","unstructured":"Jouihan, H. et al. Superior reductions in hepatic steatosis and fibrosis with co-administration of a glucagon-like peptide-1 receptor agonist and obeticholic acid in mice. Mol. Metab. 6(11), 1360\u20131370 (2017).","journal-title":"Mol. Metab."},{"issue":"5","key":"1951_CR68","doi-asserted-by":"publisher","first-page":"1334","DOI":"10.1002\/cpt.2105","volume":"109","author":"BJ Kirby","year":"2021","unstructured":"Kirby, B. J. et al. Organic anion transporting polypeptide inhibition dramatically increases plasma exposure but not pharmacodynamic effect nor inferred hepatic intracellular exposure of firsocostat. Clin. Pharmacol. Ther. 109(5), 1334\u20131341 (2021).","journal-title":"Clin. Pharmacol. Ther."},{"issue":"10","key":"1951_CR69","doi-asserted-by":"publisher","first-page":"1849","DOI":"10.1111\/liv.13706","volume":"38","author":"ZM Younossi","year":"2018","unstructured":"Younossi, Z. M. et al. Improvement of hepatic fibrosis and patient-reported outcomes in non-alcoholic steatohepatitis treated with selonsertib. Liver Int. 38(10), 1849\u20131859 (2018).","journal-title":"Liver Int."},{"issue":"1","key":"1951_CR70","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1016\/j.jhep.2020.02.027","volume":"73","author":"SA Harrison","year":"2020","unstructured":"Harrison, S. A. et al. Selonsertib for patients with bridging fibrosis or compensated cirrhosis due to NASH: Results from randomized phase III STELLAR trials. J. Hepatol. 73(1), 26\u201339 (2020).","journal-title":"J. Hepatol."},{"issue":"9","key":"1951_CR71","doi-asserted-by":"publisher","first-page":"1109","DOI":"10.1007\/s40262-020-00878-y","volume":"59","author":"CH Nelson","year":"2020","unstructured":"Nelson, C. H. et al. Pharmacokinetics, safety, and tolerability of selonsertib, an apoptosis signal-regulating kinase 1 (ASK1) inhibitor, following first-in-human single and multiple ascending doses in healthy subjects. Clin. Pharmacokinet. 59(9), 1109\u20131117 (2020).","journal-title":"Clin. Pharmacokinet."},{"key":"1951_CR72","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1016\/j.matbio.2015.05.006","volume":"47","author":"B Hinz","year":"2015","unstructured":"Hinz, B. The extracellular matrix and transforming growth factor-\u03b21: Tale of a strained relationship. Matrix Biol. 47, 54\u201365 (2015).","journal-title":"Matrix Biol."},{"issue":"5","key":"1951_CR73","doi-asserted-by":"publisher","first-page":"1090","DOI":"10.1016\/j.jhep.2013.12.025","volume":"60","author":"F Tacke","year":"2014","unstructured":"Tacke, F. & Zimmermann, H. W. Macrophage heterogeneity in liver injury and fibrosis. J. Hepatol. 60(5), 1090\u20131096 (2014).","journal-title":"J. Hepatol."},{"issue":"5","key":"1951_CR74","doi-asserted-by":"publisher","first-page":"830","DOI":"10.1002\/hep.1840040505","volume":"4","author":"A Frei","year":"1984","unstructured":"Frei, A., Zimmermann, A. & Weigand, K. The N-terminal propeptide of collagen type III in serum reflects activity and degree of fibrosis in patients with chronic liver disease. Hepatology 4(5), 830\u2013834 (1984).","journal-title":"Hepatology"},{"issue":"10","key":"1951_CR75","doi-asserted-by":"publisher","first-page":"1554","DOI":"10.1111\/liv.13402","volume":"37","author":"EMG de Vries","year":"2017","unstructured":"de Vries, E. M. G. et al. Enhanced liver fibrosis test predicts transplant-free survival in primary sclerosing cholangitis, a multi-centre study. Liver Int. 37(10), 1554\u20131561 (2017).","journal-title":"Liver Int."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-01951-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-01951-7","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-01951-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,9,12]],"date-time":"2024-09-12T17:44:26Z","timestamp":1726163066000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-01951-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,23]]},"references-count":75,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["1951"],"URL":"https:\/\/doi.org\/10.1038\/s41598-021-01951-7","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-737717\/v1","asserted-by":"object"}]},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,23]]},"assertion":[{"value":"21 July 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 November 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 November 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"22765"}}