Multiplex Analysis of Intracellular Signaling Pathways in Lymphoid Cells by Microbead Suspension Arrays

Khan, Imran; Mendoza, Sara; Rhyne, Paul; Ziman, Melanie; Tuscano, Joseph M.; Eisinger, Dominic; Kung, Hsing Jien; Luciw, Paul A.

doi:10.1074/mcp.t500032-mcp200

Cited by 27 publications

(25 citation statements)

References 58 publications

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“…Phosphoprotein Stability: the Balance between Kinases and Phosphatases: Implications for Preservation ChemistryPhosphorylation and dephosphorylation of structural and regulatory proteins are major intracellular control mechanisms (20). Protein-tyrosine phosphatases remove phosphate groups from phosphorylated tyrosine residues of proteins.…”

Section: Discussionmentioning

confidence: 99%

A Portrait of Tissue Phosphoprotein Stability in the Clinical Tissue Procurement Process

Espina

Edmiston

Heiby

et al. 2008

Molecular & Cellular Proteomics

191

194

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Section: Discussionmentioning

confidence: 99%

A Portrait of Tissue Phosphoprotein Stability in the Clinical Tissue Procurement Process

Espina

Edmiston

Heiby

et al. 2008

Molecular & Cellular Proteomics

191

194

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“…[6][7][8][9][10][11] Protein phosphorylation is a dynamic balance between two opposite events: phosphorylation by highly specific kinases and dephosphorylation by less-specific phosphatases. [12][13][14] To utilize phosphoproteins as markers of cellular signaling activity and tumor proliferation, 6,8,9 it is crucial to assess their expression levels in tissue specimens that reflect the in vivo status as accurately as possible. Elevated expression levels of phospho-AKT, phospho-ERK1/ 2, phospho-mTor, and phospho-p20S6K, for example, have been described to be associated with tamoxifen resistance in postmenopausal breast cancer patients.…”

mentioning

confidence: 99%

Preanalytical variables and phosphoepitope expression in FFPE tissue: quantitative epitope assessment after variable cold ischemic time

Vassilakopoulou

Parisi

Siddiqui

et al. 2015

Laboratory Investigation

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Individualized targeted therapies for cancer patients require accurate and reproducible assessment of biomarkers to be able to plan treatment accordingly. Recent studies have shown highly variable effects of preanalytical variables on gene expression profiling and protein levels of different tissue types. Several publications have described protein degradation of tissue samples as a direct result of delay of formalin fixation of the tissue. Phosphorylated proteins are more labile and epitope degradation can happen within 30 min of cold ischemic time. To address this issue, we evaluated the change in antigenicity of a series of phosphoproteins in paraffin-embedded samples from breast tumors as a function of time to formalin fixation. A tissue microarray consisting of 93 breast cancer specimens with documented time-to-fixation was used to evaluate changes in antigenicity of 12 phosphoepitopes frequently used in research settings as a function of cold ischemic time. Analysis was performed in a quantitative manner using the AQUA technology for quantitative immunofluorescence. For each marker, least squares univariate linear regression was performed and confidence intervals were computed using bootstrapping. The majority of the epitopes tested revealed changes in expression levels with increasing time to formalin fixation. Some phosphorylated proteins, such as phospho-HSP27 and phospho-S6 RP, involved in posttranslational modification and stress response pathways increased in expression or phosphorylation levels. Others (like phospho-AKT, phosphor-ERK1/2, phospho-Tyrosine, phospho-MET, and others) are quite labile and loss of antigenicity can be reported within 1-2 h of cold ischemic time. Therefore specimen collection should be closely monitored and subjected to quality control measures to ensure accurate measurement of these epitopes. However, a few phosphoepitopes (like phospho-JAK2 and phospho-ER) are sufficiently robust for routine usage in companion diagnostic testing.

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“…Many research groups have reported that the ZAP-70 family of tyrosine kinases are selectively retained and expressed in T cell proliferative diseases (49). A subgroup of CLL was reported to overexpress ZAP-70 thus enabling T cells to develop and differentiate through CD3-TCRmediated signaling pathways (16). Therefore, ZAP-70 has been suggested to be an excellent biomarker for prognosis in CLL (9 -11) and might be a logical target for the development of potent immunosuppressive agents for T cell-mediated diseases (5,50).…”

Section: Discussionmentioning

confidence: 99%

“…The ZAP-70 tyrosine kinase is reported to play a critical role in T cell activation and the immune response, and therefore might be a logical target for immunomodulatory therapies (5). Crespo et al (14) observed that among B cell and T cell lymphoproliferative disorders, a high level of ZAP-70 expression is found in T cell proliferative diseases, acute lymphoblastic leukemia, and a subgroup of chronic lymphocytic leukemia (CLL) (15,16). These * This work was supported, in whole or in part, by National Institutes of Health Grants CA81064, CA77646, CA111536, and CA120388.…”

mentioning

confidence: 94%

(-)-Epigallocatechin Gallate Regulates CD3-mediated T Cell Receptor Signaling in Leukemia through the Inhibition of ZAP-70 Kinase

Shim

Choi

Pugliese

et al. 2008

Journal of Biological Chemistry

105

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For thousands of years, tea has been the most widely consumed beverage in the world after water. Historically, tea has been credited with various beneficial health effects, including medicinal efficacy in the prevention and treatment of numerous diseases. Thus, longevity and good health have often been associated with the habit of drinking tea (1). Four major polyphenolic catechins are found in green tea and include (Ϫ)-epicatechin (EC), 3 (Ϫ)-epicatechin 3-gallate (ECG), (Ϫ)-epigallocatechin (EGC), and (Ϫ)-epigallocatechin 3-gallate (EGCG). A cup of green tea may contain 100 -200 mg of EGCG (2). Several investigators have reported that green tea exerts cancer preventive activity at a variety of organ sites, including skin, lung, oral cavity, esophagus, stomach, small intestine, colon, pancreas, and mammary gland (1, 3, 4). However, the mechanisms explaining the cancer preventive activity of tea and tea polyphenols are still not clearly understood.The -associated 70-kDa protein (ZAP-70) is a Syk (spleen tyrosine kinase) family tyrosine kinase, which is associated with the subunit of the T cell receptor (TCR). The ZAP-70 protein is primarily expressed in T cells and natural killer cells and plays an essential role in signaling through the T cell antigen receptor (5). The TCRs are associated with tyrosine phosphorylation of multiple proteins resulting in activation of various signaling pathways causing alterations in gene expression, increased T cell proliferation, and secretion of cytokines (6). CD3 (cluster of differentiation 3) stimulation of the T cell antigen receptor plays a role in tyrosine phosphorylation of a number of cellular substrates. An important substrate of ZAP-70 is the TCR chain, which can mediate the transduction of extracellular stimuli into cellular effector functions (7,8). ZAP-70 plays a critical role in cell surface expression of T cell antigen receptor-CD3 complex signaling during the early stages of T cell development and differentiation (9 -13). The ZAP-70 tyrosine kinase is reported to play a critical role in T cell activation and the immune response, and therefore might be a logical target for immunomodulatory therapies (5). Crespo et al. (14) observed that among B cell and T cell lymphoproliferative disorders, a high level of ZAP-70 expression is found in T cell proliferative diseases, acute lymphoblastic leukemia, and a subgroup of chronic lymphocytic leukemia (CLL) (15,16). These * This work was supported, in whole or in part, by National Institutes of Health Grants CA81064, CA77646, CA111536, and CA120388. This work was also supported by The Hormel Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

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Multiplex Analysis of Intracellular Signaling Pathways in Lymphoid Cells by Microbead Suspension Arrays

Cited by 27 publications

References 58 publications

A Portrait of Tissue Phosphoprotein Stability in the Clinical Tissue Procurement Process

A Portrait of Tissue Phosphoprotein Stability in the Clinical Tissue Procurement Process

Preanalytical variables and phosphoepitope expression in FFPE tissue: quantitative epitope assessment after variable cold ischemic time

(-)-Epigallocatechin Gallate Regulates CD3-mediated T Cell Receptor Signaling in Leukemia through the Inhibition of ZAP-70 Kinase

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