Objective: The primary aim of this study was to assess patients’ preferences between radiofrequency ablation (RFA) versus hepatic resection (HR) for the treatment of small hepatocellular carcinomas (HCC). Methods: Decision analysis was performed by using probability trade-off (PTO) technique to elicit patients’ preferences and the strength of their decisions. Results: The vast majority of the study population preferred RFA over HR (70% vs. 30%, p=0.001). Their initial choice changed if 5-year survival benefit after surgery was at least 14% superior to RFA and if the 3-year disease-free survival advantage was at least 13% better than ablation. Conclusions: The results of this study suggest that fully informed cirrhotic patients would prefer RFA if diagnosed with early stage HCC even if able to undergo surgery.

There are currently two types of intra-arterial drug-delivery system (DDS) in clinical use in the palliative treatment of primary liver cancer. The chemotherapeutic drug doxorubicin (DOX) can be formulated into a drug-in-lipiodol emulsion (LIPDOX) or a microparticulate drug-eluting bead system (DEBDOX). To facilitate development of future DDSs, we need to understand the release and local distribution of drug from these DDSs into the complex, in vivo, pathological environment. The overall aim of this project was to assess and improve understanding of the in vivo release of DOX from LIPDOX and DEBDOX and its local disposition in the liver. These processes were investigated in detail in a multisampling-site, healthy pig model and in human patients with liver cancer. The mechanisms involved in DOX disposition were studied by examining potential interactions between DOX and lipiodol and/or cyclosporine A (CsA) in pigs.   In this project, the main elimination pathway for DOX and its primary metabolite doxorubicinol (DOXol) was via bile; their extensive canalicular carrier-mediated transport (e.g. ATP-binding cassette transporters ABCB1, ABCC1, ABCC2 and ABCG2) was inhibited by CsA. CsA had no effect on the carbonyl and aldo-keto reductases responsible for the metabolism of DOX into DOXol. LIPDOX released DOX more rapidly and to a greater extent into the circulation than DEBDOX, which had only released 15% of the dose in patients after 24 hrs. The systemic exposure to DOX was lower for DEBDOX than for LIPDOX. Greater fractions of DOXol were formed in blood and bile with LIPDOX than with DEBDOX. This may have been because DOX was more widely distributed into regions with increased metabolic capacity or because of increased intracellular uptake when DOX was delivered in LIPDOX. The excipient lipiodol in the LIPDOX formulation did not interact with transporters, enzymes or membranes that would explain the increased cellular uptake of DOX. In conclusion, the release of DOX from DEBDOX is more controlled in vivo than that from LIPDOX, indicating that DEBDOX is a more robust pharmaceutical product. The formulations for future optimized DDSs should therefore be more similar to DEBDOX than to LIPDOX.

NV669 est un aminosterol dérivé de la squalamine qui a montré posséder des propriétés anti-cancéreuses. L'objectif de cette étude a été de rechercher les effets bénéfiques de NV669 sur des modèles de cancers humains pancréatiques et hépatiques et de comprendre les mécanismes cellulaires et moléculaires impliqués dans la diminution de la croissance tumorale par le traitement avec NV669. Les lignées cellulaires humaines pancréatiques (BxPC3 et MiaPaca-2) et hépatiques (HepG2 et Huh7) ont été traitées avec NV669 à différentes concentrations et à différents temps. Les résultats ont montré que NV669 inhibe la prolifération des cellules cancéreuses en induisant l'arrêt du cycle cellulaire en phase G2/M via le complexe cycline B1/Cdk1 et en induisant l'apoptose via le clivage de la caspase-8 et de PARP-1, et la fragmentation de l’ADN.De plus, nos recherches in vitro ont révélé que NV669 inhibe l’activité phosphatase de PTP1B et l’expression de FAK. NV669 affecte l’expression des molécules d’adhérence CDH-1, -2 et -3 dans les lignées BxPC3 et Huh7 qui forment des monocouches cellulaires. Cela suggère qu’en inhibant PTP1B, NV669 induirait l’apoptose.Par la suite, nos résultats in vivo ont montré que NV669 inhibe la croissance des xénogreffes pancréatiques et hépatiques tumorales avec une diminution significative de la prolifération cellulaire et une augmentation de l'apoptose des cellules tumorales. Par conséquent, nos recherches suggèrent que l’analogue de la squalamine, NV669, pourrait être un agent anti cancéreux, utilisé seul ou en association avec d’autres médicaments, dans le traitement de l’adénocarcinome pancréatique et du carcinome hépatocellulaire. / NV669 is an aminosterol derived from squalamine found to possess strong antiangiogenic and anticancer effects. The aim of this study was to investigate NV669’s beneficial effects on human pancreatic and hepatic cancer models and to understand the cellular and molecular mechanisms involved in tumor growth decrease upon treatment with NV669.Pancreatic (BxPC3, MiaPaCa-2) and hepatic (HepG2, Huh7) cancer cells were treated with NV669, and the effects on proliferation, on cell cycle and death were determined. The results showed that NV669 inhibited the viability of cancer cells, induced cell cycle arrest through the regulation of G2/M phase via a decrease in the expression of cyclin B1 and phosphorylated Cdk1 and the induction of apoptosis via cleaved caspase-8 and PARP-1 and fragmented DNA. Moreover, in vitro NV669 inhibits PTP1B activity and FAK expression. NV669 impacts on the expression of adhesion molecules CDH-1, -2 and -3 in BxPC3 and Huh7 lines that form cell monolayers. This suggests that NV669 by inhibiting PTP1B would induce apoptosis. Subsequently, our in vivo results showed that NV669 inhibited the growth of pancreatic and hepatic tumor xenografts with a significant decrease in proliferation cell and an increase of tumor cell apoptosis. Therefore, NV669 may serve as an alternative anticancer agent, used alone or in association with other medications, for the treatment of pancreatic adenocarcinoma and hepatocellular carcinoma.

Justificativa: A recidiva do Carcinoma Hepatocelular (CHC) é um fator limitante da sobrevida nos indivíduos submetidos a transplante hepático (TxH). Além do número e tamanho do tumor, invasão vascular e grau de diferenciação, compreender a biologia tumoral e os biomarcadores relacionados é um passo importante para prever a recidiva e estabelecer estratégias terapêuticas. Objetivos: Analisar a expressão e valor prognóstico do receptor do Fator de Crescimento Epidérmico (EGFr) e sua relação com outros fatores implicados na recidiva no CHC. Pacientes e métodos: Foram analisados retrospectivamente os dados de 59 pacientes cirróticos submetidos a TxH no nosso centro com diagnóstico histopatológico confirmado de CHC no período de 2001 a 2006. Resultados: Foi registrada uma sobrevida global e em 1, 3 e 5 anos de 89,3%, 74,9% e 72,4% respectivamente e uma sobrevida livre de doença em 1, 3 e 5 anos de 84,8%, 69,6% e 69,6% respectivamente. A taxa de recidiva foi de 22,03%. O EGFr foi expresso em 93,2% dos tumores, mas não apresentou relação estatisticamente significatica com risco de recidiva (p=0,147) ou outros fatores de risco. Somente a invasão vascular (p=0,046) e o tamanho > 5 cm (p=0,046) foram capazes de predizer o risco de recidiva. Conclusão: O tamanho tumoral e a invasão vascular foram os principais fatores de risco relacionados à recidiva tumoral, enquanto a expressão do EGFr, embora bastante alta, não tem relação com este risco. / Background: Recurrent Hepatocellular Carcinoma (HCC) is a significant cause of death after liver transplantation (LT). Understanding tumor biology and correlate biomarkers is an important step to predict recurrence and establish new therapies. Purpose: Analyze the expression and prognostic significance of Epidermal Growth Factor receptor (EGFr) on HCC and its relationship with other prognostic factors. Patients and methods: Data from 59 cirrhotic patients who underwent LT between 2001 and 2006 with pathological confirmation of HCC were analyzed retrospectively. Results: The overall survival at 1, 3 e 5 years was 89.3%, 74.9%, 72.4% while recurrence-free was 84.8%, 69.6, 69.6% respectively with a recurrence rate of 22%. EGFr was expressed in 93.2% of tumors, but no statistical relationship with HCC recurrence (p=0.147) were found. Only vascular invasion (p=0.046) and tumor size > 5 cm (p=0.046) predicted tumor recurrence. Conclusion: EGFr was widely expressed on our series of tumors, and, as so, has a poor prognostic value, with no significance among groups (negative, weak and strong expression) considering HCC recurrence. Only tumor size and vascular invasion were significant predictors of tumor recurrence.

Justificativa: A recidiva do Carcinoma Hepatocelular (CHC) é um fator limitante da sobrevida nos indivíduos submetidos a transplante hepático (TxH). Além do número e tamanho do tumor, invasão vascular e grau de diferenciação, compreender a biologia tumoral e os biomarcadores relacionados é um passo importante para prever a recidiva e estabelecer estratégias terapêuticas. Objetivos: Analisar a expressão e valor prognóstico do receptor do Fator de Crescimento Epidérmico (EGFr) e sua relação com outros fatores implicados na recidiva no CHC. Pacientes e métodos: Foram analisados retrospectivamente os dados de 59 pacientes cirróticos submetidos a TxH no nosso centro com diagnóstico histopatológico confirmado de CHC no período de 2001 a 2006. Resultados: Foi registrada uma sobrevida global e em 1, 3 e 5 anos de 89,3%, 74,9% e 72,4% respectivamente e uma sobrevida livre de doença em 1, 3 e 5 anos de 84,8%, 69,6% e 69,6% respectivamente. A taxa de recidiva foi de 22,03%. O EGFr foi expresso em 93,2% dos tumores, mas não apresentou relação estatisticamente significatica com risco de recidiva (p=0,147) ou outros fatores de risco. Somente a invasão vascular (p=0,046) e o tamanho > 5 cm (p=0,046) foram capazes de predizer o risco de recidiva. Conclusão: O tamanho tumoral e a invasão vascular foram os principais fatores de risco relacionados à recidiva tumoral, enquanto a expressão do EGFr, embora bastante alta, não tem relação com este risco. / Background: Recurrent Hepatocellular Carcinoma (HCC) is a significant cause of death after liver transplantation (LT). Understanding tumor biology and correlate biomarkers is an important step to predict recurrence and establish new therapies. Purpose: Analyze the expression and prognostic significance of Epidermal Growth Factor receptor (EGFr) on HCC and its relationship with other prognostic factors. Patients and methods: Data from 59 cirrhotic patients who underwent LT between 2001 and 2006 with pathological confirmation of HCC were analyzed retrospectively. Results: The overall survival at 1, 3 e 5 years was 89.3%, 74.9%, 72.4% while recurrence-free was 84.8%, 69.6, 69.6% respectively with a recurrence rate of 22%. EGFr was expressed in 93.2% of tumors, but no statistical relationship with HCC recurrence (p=0.147) were found. Only vascular invasion (p=0.046) and tumor size > 5 cm (p=0.046) predicted tumor recurrence. Conclusion: EGFr was widely expressed on our series of tumors, and, as so, has a poor prognostic value, with no significance among groups (negative, weak and strong expression) considering HCC recurrence. Only tumor size and vascular invasion were significant predictors of tumor recurrence.

La modification post-traductionnelle des chimiokines par la dipeptidyl peptidase-4 (DPP4 ou CD26) régule négativement le trafic des lymphocytes, et son inhibition améliore la migration des lymphocytes T et l'immunité anti-tumorale en préservant la forme fonctionnelle de CXCL10. En étendant ces résultats initiaux aux humains et à un modèle préclinique de carcinome hépatocellulaire, nous avons découvert un nouveau mécanisme par lequel l'inhibition de DPP4 améliore les réponses anti-tumorales par le recrutement des éosinophiles. Plus précisément, l'administration d'inhibiteurs de DPP4 (DPP4i) conduit à des concentrations tumorales plus élevées de CCL11 (ou eotaxine) et à une augmentation de la migration des éosinophiles exprimant CCR3 dans les tumeurs. Un meilleur contrôle de la croissance tumorale a été observé lors du traitement par DPP4i, un effet conservé chez les souris Rag2–/– mais abrogé uniquement lors de la déplétion des éosinophiles ou de l'inhibition de leur dégranulation. Nous avons également démontré que l'expression tumorale d’IL-33 était nécessaire et suffisante pour une réponse anti-tumorale médiée par les éosinophiles et que ce mécanisme contribuait à l'efficacité des inhibiteurs de points de contrôle immunitaires. Ces résultats révèlent un nouveau mécanisme par lequel le contrôle tumoral est médiée par IL-33 et les éosinophiles, mécanisme ici révélé lorsque les mécanismes endogènes de régulation immunitaire par DPP4 sont inhibés. / Dipeptidyl peptidase-4 (DPP4 or CD26)–mediated post-translational modification of chemokines has been shown to negatively regulate lymphocyte trafficking, and its inhibition enhances T cell migration and tumor immunity by preserving functional CXCL10. In extending these initial findings to humans and pre-clinical hepatocellular carcinoma models, we discovered a new mechanism whereby DPP4 inhibition improves anti-tumor responses by eosinophil recruitment. Specifically, administration of DPP4 inhibitors (DPP4i) resulted in higher concentrations of CCL11 (or eotaxin) and increased CCR3-mediated eosinophil migration into mouse tumors. Enhanced tumor control was observed upon treatment with DPP4i, an effect strikingly preserved in Rag2–/– mice, and abrogated only upon depletion of eosinophils or inhibition of their degranulation. We further demonstrated that tumor expression of IL-33 was necessary and sufficient for eosinophil-mediated anti-tumor responses, and that this mechanism contributed to checkpoint inhibitor efficacy. These findings provide new insight into IL-33- and eosinophil-mediated tumor control, revealed when endogenous mechanisms of DPP4 immune regulation are inhibited.

Justificativa: A recidiva do Carcinoma Hepatocelular (CHC) é um fator limitante da sobrevida nos indivíduos submetidos a transplante hepático (TxH). Além do número e tamanho do tumor, invasão vascular e grau de diferenciação, compreender a biologia tumoral e os biomarcadores relacionados é um passo importante para prever a recidiva e estabelecer estratégias terapêuticas. Objetivos: Analisar a expressão e valor prognóstico do receptor do Fator de Crescimento Epidérmico (EGFr) e sua relação com outros fatores implicados na recidiva no CHC. Pacientes e métodos: Foram analisados retrospectivamente os dados de 59 pacientes cirróticos submetidos a TxH no nosso centro com diagnóstico histopatológico confirmado de CHC no período de 2001 a 2006. Resultados: Foi registrada uma sobrevida global e em 1, 3 e 5 anos de 89,3%, 74,9% e 72,4% respectivamente e uma sobrevida livre de doença em 1, 3 e 5 anos de 84,8%, 69,6% e 69,6% respectivamente. A taxa de recidiva foi de 22,03%. O EGFr foi expresso em 93,2% dos tumores, mas não apresentou relação estatisticamente significatica com risco de recidiva (p=0,147) ou outros fatores de risco. Somente a invasão vascular (p=0,046) e o tamanho > 5 cm (p=0,046) foram capazes de predizer o risco de recidiva. Conclusão: O tamanho tumoral e a invasão vascular foram os principais fatores de risco relacionados à recidiva tumoral, enquanto a expressão do EGFr, embora bastante alta, não tem relação com este risco. / Background: Recurrent Hepatocellular Carcinoma (HCC) is a significant cause of death after liver transplantation (LT). Understanding tumor biology and correlate biomarkers is an important step to predict recurrence and establish new therapies. Purpose: Analyze the expression and prognostic significance of Epidermal Growth Factor receptor (EGFr) on HCC and its relationship with other prognostic factors. Patients and methods: Data from 59 cirrhotic patients who underwent LT between 2001 and 2006 with pathological confirmation of HCC were analyzed retrospectively. Results: The overall survival at 1, 3 e 5 years was 89.3%, 74.9%, 72.4% while recurrence-free was 84.8%, 69.6, 69.6% respectively with a recurrence rate of 22%. EGFr was expressed in 93.2% of tumors, but no statistical relationship with HCC recurrence (p=0.147) were found. Only vascular invasion (p=0.046) and tumor size > 5 cm (p=0.046) predicted tumor recurrence. Conclusion: EGFr was widely expressed on our series of tumors, and, as so, has a poor prognostic value, with no significance among groups (negative, weak and strong expression) considering HCC recurrence. Only tumor size and vascular invasion were significant predictors of tumor recurrence.

Le carcinome hépatocellulaire (CHC) présente des mutations génétiques qui altèrent les principales voies de signalisation, notamment la voie Wnt/β-caténine. En absence de mutation génétique, certains CHC peuvent montrer une activité Wnt exacerbée suite à une inactivation épigénétique d’inhibiteurs ou à une surexpression de ligands Wnts ou de ses récepteurs. De plus, le remodelage de la matrice extracellulaire favorise la progression du CHC. Nous avons montré une association entre l’activation du signal Wnt et le remodelage de la matrice extracellulaire (MEC) dans les cirrhoses et le CHC. Puis modélisé in vitro, les effets des stimuli Wnt extracellulaires sur le phénotype de cellules hépatiques, en absence de mutation de la β-caténine. En effet, les cellules HepaRG ne présentent pas de mutations de la β-caténine, de l’axine et de p53. Ainsi, la stimulation Wnt3a des cellules HepaRG induisait la formation de palissades de cellules fusiformes. De plus, les cellules traitées exprimaient des taux élevés de αSMA, COLIV, c-MYC, CK19 et LGR5 suggérant un phénotype myofibroblastique, en accord avec l’expression des marqueurs de transition épithélio-mésenchymateuse (TEM), SNAIL et TWIST. Ces données sont en faveur du rôle déterminant du microenvironnement Wnt activé dans la progression du CHC entrainant les cellules vers un phénotype progéniteur plus agressif via une TEM. En outre, l'analyse in silico de la signature transcriptomique de l'activation non mutationnelle de la voie Wnt a révélé un réseau de gènes impliqués dans le remodelage de la MEC, la TEM et la différenciation cellulaire. Les résultats suggèrent le rôle de HAPLN1 qui affecterait la migration cellulaire et l'expression des gènes de la MEC. De plus, LGR5 semble favoriser la dédifférenciation des hépatocytes. Au total, 8 gènes marqueurs obtenus in vitro ont été validés in vivo dans une série de 81 CHC humains, par qPCR et immunohistochimie en utilisant des tissus micro-array (78 CHC et 5 foies contrôles). Au total, l'ensemble des données suggère que HAPLN1 a une valeur pronostique sur la récidive et la survie globale du CHC. HAPLN1semble être indépendant du statut mutationnel la β-caténine et des variables cliniques. De plus, sa valeur pronostique est additive avec celle de CK19 + EpCAM et il semble agir en synergie avec NOG. / Hepatocellular carcinoma (HCC) displays signaling pathway disorders, including Wnt/β-catenin. Up-regulation of extracellular Wnt pathway agonists and down-regulation of extracellular Wnt pathway inhibitors result in non-mutational activation of Wnt signaling. In addition, increased extracellular matrix remodeling fosters HCC progression. Thus, we showed that enhanced Wnt signaling is associated with extracellular matrix remodeling in human cirrhosis and cancer. To further investigate non-mutational Wnt pathway activation, we established a model of Wnt activation in HepaRG human HCC progenitor cells carrying wild-type β-catenin, axin and p53. HepaRG progenitor cells treated with Wnt3a became fusiform and grew in palisades with enhanced expression of αSMA, COLIV, CK19, c-MYC, LGR5, SNAIL and TWIST, suggesting that enhanced extracellular Wnt signaling may drive HCC cells toward a more aggressive progenitor and epithelial mesenchymal transition (EMT) phenotype. Moreover, in silico analysis of the transcriptomic signature of non-mutational Wnt activation revealed a gene network involved in ECM remodeling, EMT and cell fate. Results suggest a role of HAPLN1, affecting extracellular matrix gene expression and cell migration and of LGR5 in hepatocyte dedifferentiation. Eight genes among the HepaRG gene expression dataset were validated in vivo in a collection of 81 human HCC samples and controls by qPCR and immunohistochemistry using tissue micro-arrays (78 HCC samples and 5 normal livers) in the light of β-catenin activation and mutational status. In conclusion, data suggest that HAPLN1 has a prognostic value on overall survival and recurrence of HCC. HAPLN1 appears to be independent of clinical features and β-catenin mutationnal status. Moreover, HAPLN1 appears to have an additive prognostic value with CK19 + EpCAM and act synergistically with NOG.

Philosophiae Doctor - PhD / To ameliorate Hepatitis C Virus (HCV) therapeutic and diagnostic challenges requires robust intervention strategies, including approaches that leverage the plethora of rich data published in biomedical literature to gain greater understanding of HCV pathobiological mechanisms. The multitudes of metadata originating from HCV clinical trials as well as low and high-throughput experiments embedded in text corpora can be mined as data sources for the implementation of HCV-specific resources. HCV-customized resources may support the generation of worthy and testable hypothesis and reveal potential research clues to augment the pursuit of efficient diagnostic biomarkers and therapeutic targets. This research thesis report the development of two freely available HCV-specific web-based resources: (i) Dragon Exploratory System on Hepatitis C Virus (DESHCV) accessible via http://apps.sanbi.ac.za/DESHCV/ or http://cbrc.kaust.edu.sa/deshcv/ and(ii) Hepatitis C Virus Protein Interaction Database (HCVpro) accessible via http://apps.sanbi.ac.za/hcvpro/ or http://cbrc.kaust.edu.sa/hcvpro/.DESHCV is a text mining system implemented using named concept recognition and cooccurrence based approaches to computationally analyze about 32, 000 HCV related abstracts obtained from PubMed. As part of DESHCV development, the pre-constructed dictionaries of the Dragon Exploratory System (DES) were enriched with HCV biomedical concepts, including HCV proteins, name variants and symbols to enable HCV knowledge specific exploration. The DESHCV query inputs consist of user-defined keywords, phrases and concepts. DESHCV is therefore an information extraction tool that enables users to computationally generate association between concepts and support the prediction of potential hypothesis with diagnostic and therapeutic relevance.Additionally, users can retrieve a list of abstracts containing tagged concepts that can be used to overcome the herculean task of manual biocuration. DESHCV has been used to simulate previously reported thalidomide-chronic hepatitis C hypothesis and also to model a potentially novel thalidomide-amantadine hypothesis.HCVpro is a relational knowledgebase dedicated to housing experimentally detected HCV-HCV and HCV-human protein interaction information obtained from other databases and curated from biomedical journal articles. Additionally, the database contains consolidated biological information consisting of hepatocellular carcinoma(HCC) related genes, comprehensive reviews on HCV biology and drug development,functional genomics and molecular biology data, and cross-referenced links to canonical pathways and other essential biomedical databases. Users can retrieve enriched information including interaction metadata from HCVpro by using protein identifiers,gene chromosomal locations, experiment types used in detecting the interactions, PubMed IDs of journal articles reporting the interactions, annotated protein interaction IDs from external databases, and via “string searches”. The utility of HCVpro has been demonstrated by harnessing integrated data to suggest putative baseline clues that seem to support current diagnostic exploratory efforts directed towards vimentin. Furthermore,eight genes comprising of ACLY, AZGP1, DDX3X, FGG, H19, SIAH1, SERPING1 and THBS1 have been recommended for possible investigation to evaluate their diagnostic potential. The data archived in HCVpro can be utilized to support protein-protein interaction network-based candidate HCC gene prioritization for possible validation by experimental biologists.

There are currently two types of drug formulation in clinical use in the locoregional treatment of intermediate hepatocellular carcinoma (HCC). In the emulsion LIPDOX, the cytostatic agent doxorubicin (DOX) is dissolved in the aqueous phase, which is emulsified with the oily contrast agent Lipiodol® (LIP). In the microparticular system DEBDOX, DOX is loaded into the drug-eluting entity DC Bead™. The overall aim of the thesis was to improve pharmaceutical understanding of the LIPDOX and DEBDOX formulations, in order to facilitate the future development of novel drug delivery systems. In vivo release of DOX from the formulations and the disposition of DOX and its active metabolite doxorubicinol (DOXol) were assessed in an advanced multisampling-site acute healthy pig model and in patients with HCC. The release of DOX and disposition of DOX and DOXol where further analysed using physiologically based pharmacokinetic (PBPK) and biopharmaceutical (PBBP) modelling. The combination of in vivo investigations and in silico modelling could provide unique insight into the mechanisms behind drug release and disposition. The in vivo release of DOX from LIPDOX is not extended and controlled, as it is from DEBDOX. With both formulations, DOX is released as a burst during the early phase of administration. The in vivo release of DOX from LIPDOX was faster than from DEBDOX in both pigs and patients. The release from DEBDOX was slow and possibly incomplete. The in vivo release of DOX from LIPDOX and DEBDOX could be described by using the PBBP model in combination with in vitro release profiles. The disposition of DOX and DOXol was modelled using a semi-PBPK model containing intracellular binding sites. The contrast agent Lipiodol® did not affect the hepatobiliary disposition of DOX in the pig model. The control substance used in this study, cyclosporine A, inhibited the biliary excretion of DOX and DOXol but did not alter metabolism in healthy pigs. The disposition of DOX is similar in healthy pigs and humans, which was shown by the ease of translation of the semi-PBPK pig model to the human PBBP model.