p53 immunocytochemistry and TP53 gene mutations in patients with chronic hepatitis C virus (HCV) infection.

Chronic infection with hepatitis C virus (HCV) is regarded as a risk factor for hepatocellular carcinoma (HCC), mostly in patients with liver cirrhosis. Present study aimed at evaluation of cellular expression of p53 protein, genetic TP53 changes in liver samples and anti-p53 in serum of patients with chronic hepatitis C virus infection. The expression of p53 protein were analysed by immunocytochemistry in liver biopsies from adult patients with chronic, long-lasting hepatitis C. In order to detect TP53 mutations, PCR/SSCP and sequencing were performed. Antibodies against p53 in serum were determined using enzyme immunoassay (ELISA).In two out of 14 examined patients TP53 point mutations were detected in the liver samples. In the first patient, a substitution of C to T was demonstrated in position 1 of the codon 250, resulting in substitution of proline by serine. The other patient carried a substitution of C to G in position 13274 of the intron 6. The patient carrying mutation in the codon 250 demonstrated morphological traits of liver cirrhosis and had high number of p53-immunoreactive cell nuclei in tissue. None of the patients manifested elevated titres of serum anti-p53. In the liver, significant positive correlations were disclosed between expression of p53 on one hand and grading and staging on the other. A negative correlation was disclosed between cellular expression of p53 and duration time of infection. In conclusions, genetic changes in TP53 can be detected also in non-neoplastic lesions linked to chronic HCV infection

Transcriptional regulatory networks in Hepatitis C virus_induced hepatocellular carcinoma

HCV is an epidemic affecting an estimated 160 million individuals worldwide or approximately 2.35% of the world’s population.(1) This is partly because HCV exhibits high genetic variation which thereby characterizes each region with its own genetic prevalence. Therefore, understanding the transcriptional regulatory elements that influence the progression of liver disease in the presence of HCV infection is thereby crucial for diagnostic and therapeutic purposes. Systems biology provides a road map by which these elements may be easily identified. In this study 124 microarray samples were assessed in order to determine differentially expressed genes for 4 tissue types/conditions (normal, cirrhosis, cirrhosis HCC, and HCC). Differentially expressed genes were assessed for their functional clustering and those genes were annotated with their potential transcription factors and miRNAs. Transcriptional regulatory networks were constructed to visualize each pairwise comparison between the 4 tissue types/conditions. In this study that 12 transcription factors were found to have high expression patterns amongst all 6 pairwise comparisons and these transcription factors also provide insight the conditions of the liver as it progresses through hepatic cirrhosis, hepatic steatosis, and the induction of cancer. With the plethora of miRNAs that are found in the liver, each liver condition was found to have its own signature miRNA expression pattern. In the 6 pairwise comparisons 14 miRNAs were found to have high expression patterns in all 6 pairwise comparisons and their regulation in HCC was determined as well as their impact on cellular homeostasis. Based on the findings of this study and a systematic analysis of many studies it can be concluded that as the liver progresses from cirrhosis to steatosis and eventually becoming carcinomic there are specific transcription factors regulating this transition through each stage. Whereas the condition of the liver digresses, the down-regulation of miRNAs’ expression makes the transition of the liver through each pathological stage more apparent. Therefore, an understanding of the transcriptional regulatory attributes acts as a road map to provide interference strategies in order to target the stages in the progression of HCV induced HCC

Multiple Roles of LOXL2 in the Progression of Hepatocellular Carcinoma and Its Potential for Therapeutic Targeting

LOXL2, a copper-dependent amine oxidase, has emerged as a promising therapeutic target in hepatocellular carcinoma (HCC). Increased LOXL2 expression in HCC has been linked with an aggressive phenotype and represents a poor prognostic factor. Here, we focus on the mechanisms through which LOXL2 orchestrates multiple oncogenic functions in HCC development. We performed a review of the current knowledge on the roles LOXL2 performs in the modulation of the HCC tumor microenvironment, formation of premetastatic niches, and epithelial–mesenchymal transition. We also highlighted the complex interplay between LOXL2 and hypoxia, angiogenesis, and vasculogenic mimicry in HCC. At the end of the review, we summarize the current LOXL2 inhibitors and discuss their potential in HCC precision treatment

Personalized medicine and nutrition in hepatology for preventing chronic liver disease in Mexico

Chronic liver disease is a global health issue. Patients with chronic liver disease require a fresh approach that focuses on the genetic and environmental factors that contribute to disease initiation and progression. Emerging knowledge in the fields of Genomic Medicine and Genomic Nutrition demonstrates differences between countries in terms of genetics and lifestyle risk factors such as diet, physical activity, and mental health in chronic liver disease, which serves as the foundation for the implementation of Personalized Medicine and Nutrition (PerMed-Nut) strategies. Most of the world’s populations have descended from various ethnic groupings. Mexico’s population has a tripartite ancestral background, consisting of Amerindian, European, and African lineages, which is common across Latin America’s regional countries. The purpose of this review is to discuss the genetic and environmental components that could be incorporated into a PerMed-Nut model for metabolic-associated liver disease, viral hepatitis B and C, and hepatocellular carcinoma in Mexico. Additionally, the implementation of the PerMed-Nut approach will require updated medicine and nutrition education curricula. Training and equipping future health professionals and researchers with new clinical and investigative abilities focused on preventing liver illnesses in the field of genomic hepatology globally is a vision that clinicians and nutritionists should be concerned about

Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) represents one of the most significant global health issues, given its high prevalence and the challenging nature and physiology of the liver and hepatic surgery, in its many forms. This means that the most appropriate management for HCC should incorporate a multidisciplinary approach, combining the expertise from several different specialties. This book showcases the various steps in the development, diagnosis, staging, and management of HCC and provides views and thoughts from true experts in the field. As such, it is a useful resource for any physician or surgeon, whether training or practicing, who is interested in caring for patients with HCC

Dynamics of Liver Disease in Egypt: Shifting Paradigms of a Complex Etiology.

The burden of liver disease in Egypt is exceptionally high, maintaining the highest prevalence of hepatitis C virus (HCV) worldwide, as well as rising rates of hepatocellular carcinoma (HCC). The foundation of the HCV epidemic in Egypt is generally attributed to a mass public health campaign to eliminate schistosomiasis during the 1960’s-1980’s. Questions remain regarding the precise incidence of HCV during this campaign, the future burden of chronic disease those affected will experience, and the future direction of HCV and liver disease now that this campaign has ended. This dissertation offers a series of studies designed to precisely define the nature of HCV infections and HCC in Egypt, spatially and temporally, as well as predict the future burden and impact on the Egyptian population. Specific methods included analyses of HCC case data collected from the Gharbiah Population-based Cancer Registry (GPCR) to define demographic and spatial trends in the occurrence of HCC in Egypt, in addition to a meta-analysis and the construction of two mathematical models designed to calculate historic incidence of HCV and project future HCV-related health complications. Results identified significant heterogeneity in HCC occurrence with respect to sex and district of residence. More in-depth investigation identified significant spatial clustering of HCC associated with clusters of squamous cell carcinoma of the bladder (a proxy measure for schistosomiasis burden). Meta-analysis revealed the HCV epidemic is marked by a three-way interaction between time, geographic region, and whether individuals reside in urban or rural environments. Modeling techniques confirmed the presence of a cohort effect among those affected by the public health campaign, identified by a spike in incidence among those presently aged 30-50 years. The natural history model predicted Egypt will experience significant morbidity and mortality over the next 20 years due to the HCV epidemic. Our findings highlight the significance of developing an integrated strategy for the prevention of HCV infection. Unquestionably, additional factors contributing to liver disease burden remain to be elucidated. This information is crucial and should help define the complex etiology of liver disease in Egypt, enabling policy makers to create targeted, more efficient prevention and control programs.Ph.D.Epidemiological ScienceUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/60651/1/lehmane_1.pd

Association of hepatitis C virus genotype 2 spread with historic slave trade and commerce routes in Western Africa

The hepatitis C virus genotype 2 (HCV2) is endemic in Western and Central Africa. The HCV2 evolutionary origins remain uncertain due to the paucity of available genomes from African settings. In this study, we investigated the molecular epidemiology of HCV infections in rural Guinea, Western Africa, during 2004 and 2014. Broadly reactive nested reverse transcription polymerase chain reaction (RT-PCR)-based screening of sera from 1,571 asymptomatic adults resulted in the detection of 25 (1.5 per cent; 95 per cent confidence interval 0.9-2.3) positive samples, with a median viral load of 2.54E + 05 IU/ml (interquartile range 6.72E + 05). HCV-infected persons had a median age of 47 years, and 62.5 per cent were male and 37.5 per cent were female. The full polyprotein-encoding genes were retrieved by a combination of high throughput and Sanger sequencing from 17 samples showing sufficiently high viral loads. Phylogenetic analysis and sequence distances >= 13 per cent averaged over the polyprotein genes compared to other HCV2 subtypes revealed nine previously unknown HCV2 subtypes. The time to the most recent common ancestor of the Guinean HCV2 strains inferred in a Bayesian framework was 493 years (95 per cent Highest posterior density (HPD) 453-532). Most of the Guinean strains clustered poorly by location on both the level of sampling sites within Guinea and the level of countries in the phylogenetic reconstructions. Ancestral state reconstruction provided decisive support (Bayes factor > 100) for an origin of HCV2 in Western Africa. Phylogeographic reconstructions in a Bayesian framework pointed to a radial diffusion of HCV2 from Western African regions encompassing today's countries like Ghana, Guinea Bissau, or Burkina Faso, to Central and Northern African regions that took place from the 16th century onwards. The spread of HCV2 coincided in time and space with the main historic slave trade and commerce routes, supported by Bayesian tip-association significance testing (P = 0.01). Our study confirms the evolutionary origins of HCV2 in Western Africa and provides a potential link between historic human movements and HCV2 dispersion

DEVELOPING METHODS TO DETECT DNA REPAIR DEFECTS IN A DIVERSE POPULATION

Primary liver cancer, including hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA), is one of the fastest growing cancer types. Although HCCs and CCAs are anatomically co-localized, they have distinct etiologic and genomic characteristics that varies vastly in clinical outcome and response to therapy. The Cancer Genome Atlas (TCGA) identified a subset of HCC tumors with CCA genomic features, suggesting these tumors may be a separate HCC class based on their relatedness to CCA. While multiple groups have performed molecular characterization of liver tumors in an effort to identify subtypes, few have investigated beyond gene expression and/or mutations, in an integrated HCC and CCA analysis, or in association with outcomes and liver specific processes (e.g., liver regeneration). This is particularly important because DNA repair dysfunction and liver regeneration are tightly coupled processes implicated in impaired genomic integrity and hepatocarcinogenesis. Particularly, dysregulation of these pathways may be linked to chemoresistance. Given the lack of targeted therapeutic modalities for HCC and ongoing efforts to reduce recurrence, further characterizing and subdividing HCC based on multiple pathway interactions and identification of biomarkers that are associated with repair-mediated survival represents an unmet clinical need. To address this knowledge gap, the current body of work leveraged two TCGA studies, HCC and CCA, both detailed in molecular, histological, and clinical data across multiple platforms. Through a multi-omic approach, chapter 2 of this study characterized distinct HCC subclasses utilizing an integrated TCGA HCC and CCA dataset to gain insights into biology. Chapter 3 leveraged RNA expression profiling to investigate DNA repair in association with mitotic and regenerative signatures, and clinicopathologic variables in TCGA HCC study. This work identified three molecularly distinct HCC subclasses associated with viral infection and progression-free survival. In addition, RNA-based classification of DNA repair identified heterogeneity of repair pathways in HCC tumors, and a subset of tumors with substantial disrupted liver biology and poor outcomes. Collectively, this work contributes novel findings about HCC features and repair dysfunction that dictate prognosis, and highlights the importance of developing class specific biomarkers and targeted therapies.Doctor of Philosoph