Animal models for the study of hepatitis C virus infection and replication

Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
World Journal of Gastroenterology (Impact Factor: 2.37). 06/2012; 18(23):2909-13. DOI: 10.3748/wjg.v18.i23.2909
Source: PubMed


Hepatitis C virus (HCV) hepatitis, initially termed non-A, non-B hepatitis, has become one of the leading causes of cirrhosis and hepatocellular carcinoma worldwide. With the help of animal models, our understanding of the virus has grown substantially from the time of initial discovery. There is a paucity of available animal models for the study of HCV, mainly because of the selective susceptibility limited to humans and primates. Recent work has focused modification of animals to permit HCV entry, replication and transmission. In this review, we highlight the currently available models for the study of HCV including chimpanzees, tupaia, mouse and rat models. Discussion will include methods of model design as well as the advantages and disadvantages of each model. Particular focus is dedicated to knowledge of pathophysiologic mechanisms of HCV infection that have been elucidated through animal studies. Research within animal models is critically important to establish a complete understanding of HCV infection, which will ultimately form the basis for future treatments and prevention of disease.

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    • "(2) Some of the clinically most relevant viruses infect the liver in a species-specific way, impeding the use of animal models. For example, infections with hepatitis B virus (HBV) and hepatitis C virus (HCV) remain a global burden (Guha et al., 2004; MacArthur et al., 2012). Adenovirus is another pathogen which may infect the liver and cause severe medical problems, particular in immunocompromised individuals (Hough et al., 2005). "
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    ABSTRACT: Reconstituted three-dimensional (3D) liver models obtained by engrafting hepatic cells into an extracellular matrix (ECM) are valuable tools to study tissue regeneration, drug action and toxicology ex vivo. The aim of the present study was to establish a system for the functional investigation of a viral vector in a 3D liver model composed of human HepG2 cells on a rat ECM. An adeno-associated viral (AAV) vector expressing the Emerald green fluorescent protein (EmGFP) and a short hairpin RNA (shRNA) directed against human cyclophilin b (hCycB) was injected into the portal vein of 3D liver models. Application of the vector did not exert toxic effects, as shown by analysis of metabolic parameters. Six days after transduction, fluorescence microscopy analysis of EmGFP production revealed widespread distribution of the AAV vectors. After optimization of the recellularization and transduction conditions, averages of 55 and 90 internalized vector genomes per cell in two replicates of the liver model were achieved, as determined by quantitative PCR analysis. Functionality of the AAV vector was confirmed by efficient shRNA-mediated knockdown of hCycB by 70-90%. Our study provides a proof-of-concept that a recellularized biological ECM provides a valuable model to study viral vectors ex vivo.
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    • "Although we have recently begun adding other human and hepatocyte-derived cell culture systems to the predominant Huh7-derived cell lines, non-human – particularly murine – cell lines remain a critical stepping-stone to animal model development. A series of efforts using knockout mouse cells, selectable virus replicons, and human factor transduction has recently culminated in multiple research groups achieving the complete virus life cycle in mouse cells expressing the required entry and liver factors [4], [15], [30], but animal models of HCV infection are still limited [31]. HCV mouse models relying on SCID-uPA mice supporting xenografted human liver tissue can be infected, and virus-induced pathogenesis or drug response studied, but the mice have limited life spans, lack immune systems, and are expensive to produce [32]. "
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    ABSTRACT: Hepatitis C Virus (HCV) is a serious global health problem, infecting almost 3% of the world's population. The lack of model systems for studying this virus limit research options in vaccine and therapeutic development, as well as for studying the pathogenesis of chronic HCV infection. Herein we make use of the liver-specific microRNA miR-122 to render mouse cell lines permissive to HCV replication in an attempt to develop additional model systems for the identification of new features of the virus and its life cycle. We have determined that some wild-type and knockout mouse cell lines - NCoA6 and PKR knockout embryonic fibroblasts - can be rendered permissive to transient HCV sub-genomic RNA replication upon addition of miR-122, but we did not observe replication of full-length HCV RNA in these cells. However, other wild-type and knockout cell lines cannot be rendered permissive to HCV replication by addition of miR-122, and in fact, different NCoA6 and PKR knockout cell line passages and isolates from the same mice demonstrated varying permissiveness phenotypes and eventually complete loss of permissiveness. When we tested knockdown of NCoA6 and PKR in Huh7.5 cells, we saw no substantial impact in sub-genomic HCV replication, which we would expect if these genes were inhibitory to the virus' life cycle. This leads us to conclude that along with the influence of specific gene knockouts there are additional factors within the cell lines that affect their permissiveness for HCV replication; we suggest that these may be epigenetically regulated, or modulated by cell line immortalization and transformation.
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    • "Progress in mouse genetic manipulation and the valuable information gathered from studies performed in vitro and in the clinic have facilitated the development of chimeric models where immune-deficient mice are grafted with human hepatocytes (reviewed in Chayama et al., 2011; MacArthur et al., 2012). A more recent highlight, stemming from the progress in gene manipulation, is the generation of a humanized mouse model that can be infected by HCV (Dorner et al., 2011). "
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    ABSTRACT: Hepatocellular carcinoma (HCC), the most common form of primary liver cancer is the third leading cause of cancer-related cell death in human and the fifth in women worldwide. The incidence of HCC is increasing despite progress in identifying risk factors, understanding disease etiology and developing anti-viral strategies. Therapeutic options are limited and survival after diagnosis is poor. Therefore, better preventive, diagnostic and therapeutic tools are urgently needed, in particular given the increased contribution from systemic metabolic disease to HCC incidence worldwide. In the last three decades, technological advances have facilitated the generation of genetically engineered mouse models (GEMMs) to mimic the alterations frequently observed in human cancers or to conduct intervention studies and assess the relevance of candidate gene networks in tumor establishment, progression and maintenance. Because these studies allow molecular and cellular manipulations impossible to perform in patients, GEMMs have improved our understanding of this complex disease and represent a source of great potential for mechanism-based therapy development. In this review, we provide an overview of the current state of HCC modeling in the mouse, highlighting successes, current challenges and future opportunities.
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