eprintid: 10136333 rev_number: 13 eprint_status: archive userid: 608 dir: disk0/10/13/63/33 datestamp: 2021-10-13 10:58:10 lastmod: 2022-09-09 14:32:27 status_changed: 2021-10-13 10:58:10 type: article metadata_visibility: show creators_name: Thanapirom, K creators_name: Caon, E creators_name: Papatheodoridi, M creators_name: Frenguelli, L creators_name: Al-akkad, W creators_name: Zhenzhen, Z creators_name: Vilia, MG creators_name: Pinzani, M creators_name: Mazza, G creators_name: Rombouts, K title: Optimization and validation of a novel three-dimensional co-culture system in decellularized human liver scaffold for the study of liver fibrosis and cancer ispublished: pub subjects: RFH divisions: UCL divisions: B02 divisions: C10 divisions: D17 divisions: G91 keywords: decellularized liver scaffolds; drug screening; liver fibrosis; liver cancer; sorafenib; regorafenib; STAT3; SHP-1; TGFβ1; EMT; E-cadherin; 3D in vitro model note: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). abstract: The introduction of new preclinical models for in vitro drug discovery and testing based on 3D tissue-specific extracellular matrix (ECM) is very much awaited. This study was aimed at developing and validating a co-culture model using decellularized human liver 3D ECM scaffolds as a platform for anti-fibrotic and anti-cancer drug testing. Decellularized 3D scaffolds obtained from healthy and cirrhotic human livers were bioengineered with LX2 and HEPG2 as single and co-cultures for up to 13 days and validated as a new drug-testing platform. Pro-fibrogenic markers and cancer phenotypic gene/protein expression and secretion were differently affected when single and co-cultures were exposed to TGF-β1 with specific ECM-dependent effects. The anti-fibrotic efficacy of Sorafenib significantly reduced TGF-β1-induced pro-fibrogenic effects, which coincided with a downregulation of STAT3 phosphorylation. The anti-cancer efficacy of Regorafenib was significantly reduced in 3D bioengineered cells when compared to 2D cultures and dose-dependently associated with cell apoptosis by cleaved PARP-1 activation and P-STAT3 inhibition. Regorafenib re-versed TGF-β1-induced P-STAT3 and SHP-1 through induction of epithelial mesenchymal marker E-cadherin and downregulation of vimentin protein expression in both co-cultures engrafting healthy and cirrhotic 3D scaffolds. In their complex, the results of the study suggest that this newly proposed 3D co-culture platform is able to reproduce the natural physio-pathological microenvi-ronment and could be employed for anti-fibrotic and anti-HCC drug screening. date: 2021-09-30 date_type: published official_url: https://doi.org/10.3390/cancers13194936 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1892816 doi: 10.3390/cancers13194936 lyricists_name: Pinzani, Massimo lyricists_name: Rombouts, Krista lyricists_id: MPINZ74 lyricists_id: KROMB89 actors_name: Barczynska, Patrycja actors_id: PBARC91 actors_role: owner full_text_status: public publication: Cancers volume: 13 number: 19 article_number: 4936 issn: 2072-6694 citation: Thanapirom, K; Caon, E; Papatheodoridi, M; Frenguelli, L; Al-akkad, W; Zhenzhen, Z; Vilia, MG; ... Rombouts, K; + view all <#> Thanapirom, K; Caon, E; Papatheodoridi, M; Frenguelli, L; Al-akkad, W; Zhenzhen, Z; Vilia, MG; Pinzani, M; Mazza, G; Rombouts, K; - view fewer <#> (2021) Optimization and validation of a novel three-dimensional co-culture system in decellularized human liver scaffold for the study of liver fibrosis and cancer. Cancers , 13 (19) , Article 4936. 10.3390/cancers13194936 <https://doi.org/10.3390/cancers13194936>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10136333/1/Pinzani_cancers-13-04936-v2.pdf