**Influence of Liver Fibrosis on Lobular Zonation**

**Ahmed Ghallab 1,2,\*, Maiju Myllys 1, Christian H. Holland 3,4, Ayham Zaza 1, Walaa Murad 5, Reham Hassan 1,2, Yasser A. Ahmed 6, Tahany Abbas 5, Eman A. Abdelrahim 5, Kai Markus Schneider 7, Madlen Matz-Soja 8, Jörg Reinders 1, Rolf Gebhardt 8, Marie-Luise Berres 7, Maximilian Hatting 7, Dirk Drasdo 1,9, Julio Saez-Rodriguez 3,4, Christian Trautwein <sup>7</sup> and Jan G. Hengstler 1,\***


Received: 5 November 2019; Accepted: 28 November 2019; Published: 2 December 2019

**Abstract:** Little is known about how liver fibrosis influences lobular zonation. To address this question, we used three mouse models of liver fibrosis, repeated CCl4 administration for 2, 6 and 12 months to induce pericentral damage, as well as bile duct ligation (21 days) and mdr2−/<sup>−</sup> mice to study periportal fibrosis. Analyses were performed by RNA-sequencing, immunostaining of zonated proteins and image analysis. RNA-sequencing demonstrated a significant enrichment of pericentral genes among genes downregulated by CCl4; vice versa, periportal genes were enriched among the upregulated genes. Immunostaining showed an almost complete loss of pericentral proteins, such as cytochrome P450 enzymes and glutamine synthetase, while periportal proteins, such as arginase 1 and CPS1 became expressed also in pericentral hepatocytes. This pattern of fibrosis-associated 'periportalization' was consistently observed in all three mouse models and led to complete resistance to hepatotoxic doses of acetaminophen (200 mg/kg). Characterization of the expression response identified the inflammatory pathways TGFβ, NFκB, TNFα, and transcription factors NFKb1, Stat1, Hif1a, Trp53, and Atf1 among those activated, while estrogen-associated pathways, Hnf4a and Hnf1a, were decreased. In conclusion, liver fibrosis leads to strong alterations of lobular zonation, where the pericentral region adopts periportal features. Beside adverse consequences, periportalization supports adaptation to repeated doses of hepatotoxic compounds.

**Keywords:** zonation; liver lobule; chronic liver disease; cytochrome P450; inflammation; bile duct ligation; acetaminophen
