Chronic Liver Disease (CLD) and Acute Liver Failure (ALF) are serious medical syndromes. Current therapeutic options consist of managing complications, and liver transplant. Even if liver transplantation is thought to be suitable for CLD or ALF patients, there are not enough organs available and thus increasingly more deaths occur on the transplant waiting list. Therefore, there is a pressing need to develop additional therapies. This thesis firstly systematically reviews trials in autologous cell therapies as possible treatments for patients with cirrhosis. The published literature is imperfect and the difference in trial design means it has not been possible to conduct a meta-analysis. Regardless of these shortcomings, cell therapy is a potentially positive prospect. In ALF and CLD monocyte-macrophages have diverse roles within the liver. Monocyte and immune cell changes in ALF are investigated and it is demonstrated for the first time that patients with paracetamol induced ALF have a significantly altered blood compartment and that these changes correlate with patient outcome. It is possible that these results may help stratify which patients may spontaneously survive and which patients may require an emergency liver transplant. Furthermore, modulation of these changes may improve outcomes for patients. The thesis also examines monocyte-macrophages in cirrhotic patients and demonstrates the feasibility of differentiating cirrhotic patients’ monocytes into functional macrophages, comparable to healthy volunteers in a Good Manufacturing Practice (GMP) environment. A first in-man trial using macrophages infused to patients with cirrhosis as a potential new treatment is also detailed. Finally, this thesis outlines developmental work for cell therapy in patients with cirrhosis in the multi-centre REALISTIC trial. Patients were randomly assigned to receive; standard medical care, Granulocyte Stimulating Factor (GCSF) injections alone or GCSF combined with repeated stem cell infusion.

L’ADN méthyltransférase 3a (DNMT3A) relie environnement et phénotype par la méthylation des dinucléotides CpG, qu’on les trouve en particulier dans les régions promotrices des gènes. Hypométhylation de ces CpG est associée à l’activation de la transcription, qui permet le contrôle de l'expression génique dans des états physiologiques et pathologiques. La plupart de nos connaissances sur l’implication de Dnmt3a en pathologie concernent le cancer, quelques données montrent sa contribution à d’autres pathologies. L’athérosclérose est la maladie cardiovasculaire la plus fréquente. Plusieurs facteurs de risque contribuant à son apparition, sont liés à L’environnement. En particulier, les dyslipidémies, largement influencées par le régime alimentaire. Par ailleurs, d’abondantes données décrivent la contribution des cellules inflammatoires à la physiopathologie de cette maladie. Jusqu'à présent, un nombre croissant de données suggère un rôle de la méthylation de l’ADN dans l'athérosclérose, mais à ce jour, le rôle de Dnmt3a dans la régulation du cholestérol et le développement initial des plaques n'a pas été étudié.Nos résultats suggèrent que l’inactivation de Dnmt3a dans les monocytes/macrophages ne modifie pas le développement initial des plaques d’athérome et n’a pas d’influence sur la polarisation des macrophages in vitro. En parallèle, nous avons démontré que l’inactivation de Dnmt3a dans les hépatocytes conduit à une différence significative de cholestérolémie plasmatique qui n’est pas liée à une dérégulation des gènes majeurs impliqués dans le métabolisme du cholestérol. En revanche, nous avons mis en évidence une activation des réponses inflammatoires. / DNA methyltransferase 3a (DNMT3A) links environment to phenotypes via catalysis of CpG dinucleotides, notably found in genes promoter regions, methylation and whose hypomethylation is associated with gene transcriptional activation thus enabling the control of gene expression in physiologic and pathologic states. Most of our knowledge about its’ role in disease occurrence are based on articles demonstrating its’ implication in human cancers. Limited data from mouse studies illustrates its’ contribution to certain pathologies. Atherosclerosis constitutes the single most important contributor to the growing burden of cardiovascular disease. Risk factors contribute to disease occurrence, where most are related to environmental influences, notably Dyslipidaemia, a key initiator of atherosclerosis. Abundant data link hypercholesterolemia to atherogenesis, on the other hand, contribution of inflammatory mechanisms that couple dyslipidaemia to atheroma formation has been also appreciated. So far, a growing number of data suggests a role of Dnmt3a in atherosclerosis but to date, its role in cholesterol regulation and early plaque formation has not been clearly elucidated. Our results suggested that deletion of Dnmt3a in monocyte/macrophages does not affect the formation of early atherosclerostic plaque nor does it impact the polarization of macrophages in vitro. In parallel, we have also demonstrated that the deletion of Dnmt3a in hepatocytes leads to significant elevation in TC levels. We were not able to relate this elevation to dysregulation of major genes involved in Cholesterol regulation. On the other hand, we noticed activation of hepatic inflammatory responses.

Hepatitis C virus (HCV) induces a high rate of chronic infection via dysregulation of host immunity. We have previously shown that T-cell immunoglobulin and mucin domain protein-3 (Tim-3) is up-regulated on monocyte/macrophages (M/Mφ) during chronic HCV infection; little is known, however, about the transcription factor that controls its expression in these cells. In this study, we investigated the role of transcription factor, T-box expressed in T cells (T-bet), in Tim-3 expression in M/Mφ in the setting of HCV infection. We demonstrate that T-bet is constitutively expressed in resting CD14+ M/Mφ in the peripheral blood. M/Mφ from chronically HCV-infected individuals exhibit a significant increase in T-bet expression that positively correlates with an increased level of Tim-3 expression. Up-regulation of T-bet is also observed in CD14+ M/Mφ incubated with HCV+ Huh7.5 cells, as well as in primary M/Mφ or monocytic THP-1 cells exposed to HCV core protein in vitro, which is reversible by blocking HCV core/gC1qR interactions. Moreover, the HCV core-induced up-regulation of T-bet and Tim-3 expression in M/Mφ can be abrogated by incubating the cells with SP600125 – an inhibitor for the c-Jun N-terminal kinase (JNK) signalling pathway. Importantly, silencing T-bet gene expression decreases Tim-3 expression and enhances interleukin-12 secretion as well as signal transducer and activator of transcription 1 phosphorylation. These data suggest that T-bet, induced by the HCV core/gC1qR interaction, enhances Tim-3 expression via the JNK pathway, leading to dampened M/Mφ function during HCV infection. These findings reveal a novel mechanism for Tim-3 regulation via T-bet during HCV infection, providing new targets to combat this global epidemic viral disease.