2D-DIGE as a Strategy To Identify Serum Markers for the Progression of Prostate Cancer

Byrne, J.; Downes, Michelle; O’Donoghue, Niamh; O’Keane, Conor; O’Neill, Amanda; Fan, Yue; Fitzpatrick, John M.; Dünn, Michael J.; Watson, R. William G.

doi:10.1021/pr800570s

Cited by 106 publications

(114 citation statements)

References 62 publications

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“…The method chosen was 2-dimensional difference gel electrophoresis (2D-DIGE) because of its quantification strengths and proven success in small discovery cohorts. 8,9 Two dimensional DIGE is also capable of indicating posttranslational modifications based on isoelectric point shifting. This screening tool that facilitates the dissection of the serum proteome 10 identified leucine-rich ␣2-glycoprotein (LRG) as being overexpressed in asymptomatic patients with elevated BNP, who are at risk of development of HF.…”

Section: Clinical Perspective On P 197mentioning

confidence: 99%

Proteomic Analysis of Coronary Sinus Serum Reveals Leucine-Rich α2-Glycoprotein as a Novel Biomarker of Ventricular Dysfunction and Heart Failure

Watson

Ledwidge

Phelan

et al. 2011

Circ: Heart Failure

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Background-Heart failure (HF) prevention strategies require biomarkers that identify disease manifestation. Increases in B-type natriuretic peptide (BNP) correlate with increased risk of cardiovascular events and HF development. We hypothesize that coronary sinus serum from a high BNP hypertensive population reflects an active pathological process and can be used for biomarker exploration. Our aim was to discover differentially expressed disease-associated proteins that identify patients with ventricular dysfunction and HF. Methods and Results-Coronary sinus serum from 11 asymptomatic, hypertensive patients underwent quantitative differential protein expression analysis by 2-dimensional difference gel electrophoresis. Proteins were identified using mass spectrometry and then studied by enzyme-linked immunosorbent assay in sera from 40 asymptomatic, hypertensive patients and 105 patients across the spectrum of ventricular dysfunction (32 asymptomatic left ventricular diastolic dysfunction, 26 diastolic HF, and 47 systolic HF patients). Leucine-rich ␣2-glycoprotein (LRG) was consistently overexpressed in high BNP serum. LRG levels correlate significantly with BNP in hypertensive, asymptomatic left ventricular diastolic dysfunction, diastolic HF, and systolic HF patient groups (PՅ0.05). LRG levels were able to identify HF independent of BNP. LRG correlates with coronary sinus serum levels of tumor necrosis factor-␣ (Pϭ0.009) and interleukin-6 (Pϭ0.021). LRG is expressed in myocardial tissue and correlates with transforming growth factor-␤R1 (PϽ0.001) and ␣-smooth muscle actin (Pϭ0.025) expression. Conclusions-LRG was identified as a serum biomarker that accurately identifies patients with HF. Multivariable modeling confirmed that LRG is a stronger identifier of HF than BNP and this is independent of age, sex, creatinine, ischemia, ␤-blocker therapy, and BNP. (Circ Heart Fail. 2011;4:188-197.)Key Words: heart failure Ⅲ hypertension Ⅲ natriuretic peptides Ⅲ leucine-rich ␣2-glycoprotein A lthough therapies for heart failure (HF) have improved, it is clear that effective prevention strategies will have the most important impact on the concerning epidemiology of this syndrome. In this regard, it is essential to complement effective risk factor control with the earliest possible identification of those at risk for progressive ventricular dysfunction and HF. Hypertensive heart disease (HHD) is a welldescribed example of progressive ventricular dysfunction. Currently, it is not known why a certain proportion of patients with hypertension develop diastolic dysfunction (DD) and others do not. Furthermore, the early diagnosis of the transition to HF is critical and can be clinically challenging. A better understanding of the pathophysiological signals at play may allow for more precise diagnostic tests to define the onset of HF. This natural history is not simply explained by the degree of blood pressure control and probably involves complex disease mechanisms still not fully understood. The ability to identify which asymptom...

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Section: Clinical Perspective On P 197mentioning

confidence: 99%

Proteomic Analysis of Coronary Sinus Serum Reveals Leucine-Rich α2-Glycoprotein as a Novel Biomarker of Ventricular Dysfunction and Heart Failure

Watson

Ledwidge

Phelan

et al. 2011

Circ: Heart Failure

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“…Some studies of 2D-DIGE-based proteomics focused to identify novel clinically relevant proteins in PCa (24)(25)(26). In the present study, compared with normal prostate epithelial cells, eleven differentially expressed proteins were identified as potential candidates associated with prostate cancer metastases.…”

Section: Discussionmentioning

confidence: 81%

Quantitative proteomic study of human prostate cancer cells with different metastatic potentials

Wang

et al. 2016

International Journal of Oncology

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Abstract. Metastatic dissemination is a feature of most cancers including prostate cancer (PCa), and is the main cause of treatment failure and mortality. The aim of the study is to explore the mechanisms of PCa metastasis and to search for potential prognostic markers using proteomics. Two-dimensional fluorescent differential gel electrophoresis (2D-DIGE) was used to quantify proteins in normal prostate epithelial cells, bone metastasis-derived PC-3 cells, and visceral metastasis-derived PC-3M cells. Metastatic potential was confirmed by flow cytometry, electron microscopy, proliferating cell nuclear antigen assay, and wound healing assay. Differential protein expression was compared between PCa cells with different metastatic potentials (LNcap, DU145, PC-3 and PC-3M) and normal prostate epithelial cells (RWPE-1). Selected candidate proteins in human prostate tissues were analyzed using GOA, UniProt and GeneCards analyses. Eighty-six proteins were differentially expressed between cell lines (>1.5-fold, P<0.05). Among them, twelve proteins were identified by MALDI-TOF-MS. One protein was upregulated in normal prostate epithelial cells, nine proteins were upregulated in PC-3, and two proteins were upregulated in PC-3M. Proteins were divided into five groups according to their functions. The SETDB1 protein was closely associated with the prognosis of PCa. Bioinformatics suggested that SETDB1 might promote PCa bone metastasis through the WNT pathway. In conclusion, SETDB1 might be associated with the development of bone metastases from PCa. Further study is necessary to assess its exact role in PCa. IntroductionProstate cancer (PCa) is the most common malignancy in males and the second cause of cancer-related death in the USA and Europe (1). The incidence and mortality of PCa are rapidly increasing in China because of changes in diet and aging of the population (2). The proportion of PCa diagnosed at its early stage has increased because of the widespread screening of serum prostate-specific antigen (PSA) in the last two decades (3). However, many patients still present metastases at diagnosis, and metastases are the first factor leading to death from PCa (4). Patients with metastases do not benefit from radical prostatectomy or radiotherapy (5,6). Therefore, novel treatment approaches are needed.Proteomics is a powerful and effective tool to evaluate protein profiles (7). Two-dimensional fluorescent differential

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“…Two-dimensional gel electrophoresis was performed as previously described (22). The stained gels were scanned using a GS-800 Densitometer (Bio-Rad) and images analyzed using Progenesis SameSpots software (Nonlinear Dynamics).…”

Section: Two-dimensional Electrophoresismentioning

confidence: 99%

Bruton's Tyrosine Kinase Is Required for Apoptotic Cell Uptake via Regulating the Phosphorylation and Localization of Calreticulin

Byrne

Gabhann

Stacey

et al. 2013

The Journal of Immunology

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In addition to regulating B cell development and activation, Bruton’s tyrosine kinase (Btk) functions downstream of multiple TLRs, including TLR7, to regulate innate immune responses in myeloid cells. Although critical for defense against RNA viruses such as influenza and Sendai virus, recognition of self-RNA by TLR7 also has been shown to be an important contributor to the pathophysiology of systemic lupus erythematosus. To date, the role of Btk in regulating TLR7-mediated responses is poorly understood. In the current study, we have demonstrated a hitherto undiscovered role for Btk in apoptotic cell uptake, identifying the molecular chaperone calreticulin (CRT) as a novel substrate for Btk in regulating this response. CRT together with the transmembrane receptor CD91 function at the cell membrane and regulate uptake of C1q-opsonised apoptotic cells. Our results show that Btk directly phosphorylates CRT and that in the absence of Btk, CRT fails to localize with CD91 at the cell surface and at the phagocytic cup. Critically, a blocking Ab against CRT in wild-type macrophages mimics the inability of Btk-deficient macrophages to phagocytose apoptotic cells efficiently, indicating the critical importance of Btk in regulating CRT-driven apoptotic cell uptake. Our data have revealed a novel regulatory role for Btk in mediating apoptotic cell clearance, with CRT identified as the critical component of the CRT/CD91/C1q system targeted by Btk. Given the importance of clearing apoptotic cell debris to prevent inappropriate exposure of TLRs to endogenous ligands, our results have important implications regarding the role of Btk in myeloid cell function.

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2D-DIGE as a Strategy To Identify Serum Markers for the Progression of Prostate Cancer

Cited by 106 publications

References 62 publications

Proteomic Analysis of Coronary Sinus Serum Reveals Leucine-Rich α2-Glycoprotein as a Novel Biomarker of Ventricular Dysfunction and Heart Failure

Proteomic Analysis of Coronary Sinus Serum Reveals Leucine-Rich α2-Glycoprotein as a Novel Biomarker of Ventricular Dysfunction and Heart Failure

Quantitative proteomic study of human prostate cancer cells with different metastatic potentials

Bruton's Tyrosine Kinase Is Required for Apoptotic Cell Uptake via Regulating the Phosphorylation and Localization of Calreticulin

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