Proteomics for hepatocellular carcinoma marker discovery
ABSTRACT Refinements of serological markers and screening of patients at high risk for developing hepatocellular carcinoma (HCC) may lead to better HCC detection, earlier intervention, and successful treatment, improving long-term outcomes. Proteomics promises the discovery of biomarkers for early HCC detection and diagnosis. Proteomic-based profiling uniquely allows delineation of global changes in expression patterns resulting from transcriptional and posttranscriptional control, posttranslational modifications, and shifts in proteins between cellular compartments. Approaches to that effect include direct serum protein profiling and comparative analysis of protein expression in normal, precancerous, and early-stage tumor tissues. Identification of panels of tumor antigens that elicit a humoral response also may contribute to the discovery of new markers for HCC screening and diagnosis. Today, 2-dimensional polyacrylamide gel electrophoresis, multidimensional liquid chromatography, mass spectrometry, and protein microarrays are among the proteomic tools available for biomarker and drug target discovery. We review these technologies and their application to the study of HCC. Our objective is to provide a framework for appreciating the promise, while at the same time understanding the challenges behind translating proteomics discovery into novel diagnostic tests.
- SourceAvailable from: Suzan M Semaan
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- "Currently SELDI have been used for evaluating serum fi nding it useful for the discovery and identifi cation of potential biomarkers (Li et al. 2002; Petricoin et al. 2002a; Qu et al. 2002; Petricoin et al. 2002b; Petricoin et al. 2002c; Issaq et al. 2002; Miguet et al. 2006). Although limitations during large profi ling experiments, where some observations noted that spectra can vary based on analytical factors (such as the time of processing, issues in the reproducibility of SELDI (Petricoin et al. 2002a; Deng et al. 2003; Diamandis, 2003), and usually unidentifi ed individual proteins (Chignard and Beretta, 2004)), there are advantages that include prior to analysis removal of components (salts or detergents) that commonly cause problems with other analytical tools, thus leaving only those proteins that are actively interacting with the spot surfaces to be analyzed in the ProteinChip Reader (Romer et al. 2002). "
ABSTRACT: Biomarkers are biomolecules that serve as indicators of biological and pathological processes, or physiological and pharmacological responses to a drug treatment. Because of the high abundance of albumin and heterogeneity of plasma lipoproteins and glycoproteins, biomarkers are difficult to identify in human serum. Due to the clinical significance the identification of disease biomarkers in serum holds great promise for personalized medicine, especially for disease diagnosis and prognosis. This review summarizes some common and emerging proteomics techniques utilized in the separation of serum samples and identification of disease signatures. The practical application of each protein separation or identification technique is analyzed using specific examples. Biomarkers of cancers of prostate, breast, ovary, and lung in human serum have been reviewed, as well as those of heart disease, arthritis, asthma, and cystic fibrosis. Despite the advancement of technology few biomarkers have been approved by the Food and Drug Administration for disease diagnosis and prognosis due to the complexity of structure and function of protein biomarkers and lack of high sensitivity, specificity, and reproducibility for those putative biomarkers. The combination of different types of technologies and statistical analysis may provide more effective methods to identify and validate new disease biomarkers in blood.Biomarker insights 02/2007; 2:21-43.
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ABSTRACT: A recent theory stipulates that during the course of HIV infection, there is a shift in immune response from T-helper 1 to T-helper 2 responses, characterised by elevated secretions of relevant cytokines. Cytokine profiles of 15 asymptomatic (treatment naïve) and 26 symptomatic (undergoing treatment) HIV-1 patients was determined to investigate the validity of this theory. HIV-1 RNA was quantified using the COBAS TaqMan HIV-1 test, CD4 T-cell counts with the FACSCalibur flow cytometer and IL-1, IL-4, IL-6, IL-10 and IFN-gamma cytokine levels by ELISA method. The asymptomatic group had significantly higher RNA levels (p-value; 0.000006) and lower CD4 T-cell counts than the symptomatic group indicating ongoing disease progression in the absence of antiretroviral treatment and a positive response to HIV treatment by the symptomatic group. IL-1, IL-4 and IFN-gamma were undetectable in most study subjects. IL-10 and IL-6 levels was relatively lower in the asymptomatic group (mean value; 206.352 pg/ml, 10.516 pg/ml) than the symptomatic group (mean value; 417.539, 18.387 pg/ml). Lower levels of proinflammatory cytokines (IL-1, IFN-gamma) in both study groups and elevated levels of anti-inflammatory cytokine IL-10, confirms that there is a shift in immune response as HIV infection progress to AIDS. In addition, the presence of a progressive trend of anti-inflammatory cytokine, IL-10 and proinflammatory cytokine, IL-6 in 12 symptomatic patients tested 3 months after antiretroviral therapy indicates an attempt by antiretrovirals to restore immune function.Roumanian archives of microbiology and immunology 01/2010; 69(1):24-34.
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ABSTRACT: There is an emerging trend in immunotoxicological studies to use the multiplex technologies for testing the safety and the efficacy of new pharmaceuticals by using cytokines profiling as biomarker. The Luminex 200 xMAP (multi-analyte profiling) technology provides simultaneous measurement of multiple cytokines in small sample volumes, expressing rapidly the differences between various test compounds. The aim is to develop and validate the Luminex 200 multiplex immunoassays by correlation with ELISA (enzyme-linked immunosorbent assays) for implementation in evaluating cytokine profiling in immunotoxicological studies in vitro. METHODS: Human peripheral whole blood from healthy subject diluted 1+4 with RPMI 1640 was cultured 48 hours in 28 experimental variants: control, in presence of mitogens, bioflavonoid extracts (from Crataegus monogyna and Echinacea purpurea) as cytoprotectors and with a toxic compound [Pb(NO3)2]), separately or variously combined. IL-1beta and IL-2 were comparatively performed by xMAP and ELISA immunoassays from the same sample to initialize validation of multiplex cytokine panel: IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-alpha, IFN-gamma, usually performed by Luminex 200 system in our immunotoxicological studies. The results indicate similarly typed trends of cytokine values obtained by both methods, with comparable relative changes in presence of mitogens, bioflavonoids and toxic, respectively. Although xMAP absolute cytokine values were higher than ELISA values, the correlation between multiplexed assay and ELISA was good for IL-1beta and IL-2 with positive correlation coefficients near to 1. Conclusions. Quantitative differences between absolute values for IL-1beta and IL-2 obtained by xMAP and ELISA assays are found, but the relative values are comparable and the two methods keep similar trends in similar exposure conditions. The performance parameters of the xMAP assay and the good correlation coefficients with the "gold standard" ELISA recommend to validate the multiplex assay for analyzing cytokine profiles in immunotoxicological studies in vitro.Roumanian archives of microbiology and immunology 01/2010; 69(1):13-9.