{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T18:17:51Z","timestamp":1790792271118,"version":"4.1.0"},"posted":{"date-parts":[[2023,11,1]]},"group-title":"Cell Biology","reference-count":44,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,11,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>\n                    Precision-Cut Liver Slices (PCLS) are an\n                    <jats:italic>ex vivo<\/jats:italic>\n                    culture model developed to study hepatic drug metabolism. One of the main benefits of this model is that it retains the structure and cellular composition of the native liver. PCLS also represents a potential model system to study liver fibrosis in a setting that more closely approximates\n                    <jats:italic>in vivo<\/jats:italic>\n                    pathology than\n                    <jats:italic>in vitro<\/jats:italic>\n                    methods. The aim of this study was to assess whether responses to antifibrotic interventions can be detected and quantified with PCLS.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    PCLS of 250 \u03bcm thickness were prepared from four different murine fibrotic liver models: choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD), thioacetamide (TAA), diethylnitrosamine (DEN), and carbon tetrachloride (CCl\n                    <jats:sub>4<\/jats:sub>\n                    ). PCLS were treated with 5 \u03bcM Erlotinib for 72 hours. Histology and gene expression were then compared with\n                    <jats:italic>in vivo<\/jats:italic>\n                    murine experiments and TGF-\u03b21 activated hepatic stellate cells (HSCs). These types of PCLS characterization were also evaluated in PCLS from human cirrhotic liver.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    PCLS viability in culture was stable for 72 hours. Treatment of erlotinib, an EGFR inhibitor significantly inhibited the expression of profibrogenic genes\n                    <jats:italic>Il6<\/jats:italic>\n                    ,\n                    <jats:italic>Col1a1<\/jats:italic>\n                    and\n                    <jats:italic>Timp1<\/jats:italic>\n                    in PCLS from CDAHFD-induced cirrhotic mice, and\n                    <jats:italic>Il6<\/jats:italic>\n                    ,\n                    <jats:italic>Col1a1<\/jats:italic>\n                    and\n                    <jats:italic>Tgfb1<\/jats:italic>\n                    in PCLS from TAA-induced cirrhotic rats. Erlotinib treatment of PCLS from DEN-induced cirrhotic rats inhibited the expression of\n                    <jats:italic>Col1a1<\/jats:italic>\n                    ,\n                    <jats:italic>Timp1<\/jats:italic>\n                    ,\n                    <jats:italic>Tgfb1<\/jats:italic>\n                    and\n                    <jats:italic>Il6<\/jats:italic>\n                    , which was consistent with the impact of erlotinib on\n                    <jats:italic>Col1a1<\/jats:italic>\n                    and\n                    <jats:italic>Tgfb1<\/jats:italic>\n                    expression in\n                    <jats:italic>in vivo<\/jats:italic>\n                    DEN-induced cirrhosis. Erlotinib treatment of PCLS from CCl\n                    <jats:sub>4<\/jats:sub>\n                    -induced cirrhosis caused reduced expression of\n                    <jats:italic>Timp1<\/jats:italic>\n                    ,\n                    <jats:italic>Col1a1<\/jats:italic>\n                    and\n                    <jats:italic>Tgfb1<\/jats:italic>\n                    , which was consistent with the effect of erlotinib in\n                    <jats:italic>in vivo<\/jats:italic>\n                    CCl\n                    <jats:sub>4<\/jats:sub>\n                    -induced cirrhosis. In addition, in HSCs at PCLS from normal mice, TGF-\u03b21 treatment upregulated\n                    <jats:italic>Acta2<\/jats:italic>\n                    (\n                    <jats:italic>\u03b1SMA<\/jats:italic>\n                    ), while treatment with erlotinib inhibited the expression of\n                    <jats:italic>Acta2<\/jats:italic>\n                    . Similar expression results were observed in TGF-\u03b21 treated\n                    <jats:italic>in vitro<\/jats:italic>\n                    HSCs. Expression of MMPs and TIMPs, key regulators of fibrosis progression and regression, were also significantly altered under erlotinib treatment in PCLS. Expression changes under erlotinib treatment were also corroborated with PCLS from human cirrhosis samples.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>\n                    The responses to antifibrotic interventions can be detected and quantified with PCLS at the gene expression level. The antifibrotic effects of erlotinib are consistent between PCLS models of murine cirrhosis and those observed\n                    <jats:italic>in vivo<\/jats:italic>\n                    and\n                    <jats:italic>in vitro<\/jats:italic>\n                    . Similar effects were also reproduced in PCLS derived from patients with cirrhosis. PCLS is an excellent model to assess antifibrotic therapies that is aligned with the principles of Replacement, Reduction and Refinement (3Rs).\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.10.30.564772","type":"posted-content","created":{"date-parts":[[2023,11,2]],"date-time":"2023-11-02T01:20:11Z","timestamp":1698888011000},"source":"Crossref","is-referenced-by-count":5,"title":["Precision-Cut Liver Slices as an\n                  <i>ex vivo<\/i>\n                  model to evaluate antifibrotic therapies for liver fibrosis and cirrhosis"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5640-4720","authenticated-orcid":false,"given":"Yongtao","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ben","family":"Leaker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guoliang","family":"Qiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mozhdeh","family":"Sojoodi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ibrahim Ragab","family":"Eissa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eliana T.","family":"Epstein","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan","family":"Eddy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oizoshimoshiofu","family":"Dimowo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georg M.","family":"Lauer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raymond T.","family":"Chung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Motaz","family":"Qadan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Lanuti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bryan C.","family":"Fuchs","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenneth K.","family":"Tanabe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"issue":"9963","key":"2023110820151089000_2023.10.30.564772v2.1","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/S0140-6736(14)61682-2","volume":"385","author":"Global, regional, and national age\u2013sex specific all-cause and cause-specific mortality for 240 causes of death, 1990\u20132013: a systematic analysis for the Global Burden of Disease Study 2013","year":"2015","journal-title":"The Lancet"},{"key":"2023110820151089000_2023.10.30.564772v2.2","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2018.12.036"},{"key":"2023110820151089000_2023.10.30.564772v2.3","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(16)31012-1"},{"key":"2023110820151089000_2023.10.30.564772v2.4","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(16)31678-6"},{"key":"2023110820151089000_2023.10.30.564772v2.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.cld.2015.01.001"},{"key":"2023110820151089000_2023.10.30.564772v2.6","doi-asserted-by":"publisher","DOI":"10.1002\/hep.25670"},{"key":"2023110820151089000_2023.10.30.564772v2.7","doi-asserted-by":"publisher","DOI":"10.1002\/hep.30651"},{"key":"2023110820151089000_2023.10.30.564772v2.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2006.04.009"},{"key":"2023110820151089000_2023.10.30.564772v2.9","doi-asserted-by":"crossref","unstructured":"Hubrecht, R. 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