Clinical-translational strategies for the elevation of Nm23-H1 metastasis suppressor gene expression

Marshall, Jean‐Claude; Lee, Jong Heun; Steeg, Patricia S.

doi:10.1007/s11010-009-0116-3

Cited by 17 publications

(12 citation statements)

References 33 publications

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“…The presence of high concentrations NM23-H1 has been shown to be an indicator of prognosis in many cancers17, 20–27, with its highest concentration in the non-tumorigenic state and a significantly reduced level in the metastatic state17. Over-expression of NM23-H1 can eradicate tumor cell motility and invasion28 while promoting cellular differentiation, and shuts down anchorage-independent growth29.…”

Section: Resultsmentioning

confidence: 99%

Mass Spectrometry Detection of Histidine Phosphorylation on NM23-H1

2010

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Phosphorylation is a ubiquitous protein post-translational modification that is intimately involved in most aspects of cellular regulation. Currently, most proteomic analyses are performed with phosphorylation searches for serine, threonine and tyrosine modifications, as the phosphorylated residues of histidine and aspartic acid are acid labile, and thus undetectable with most proteomic methodologies. Here, we present a novel buffer system to show histidine phosphorylation of NM23-H1, the product of the first identified putative human metastasis suppressor gene (NME1), which catalyzes the transfer of the γ-phosphate from nucleoside triphosphates to nucleoside diphosphates. Based on a pH titration of LC elution buffers and MS/MS identification, recombinant NM23-H1 subjected to auto-phosphorylation was shown to contain phosphorylated histidine at residue 118 at pH 5 and pH 6, with each level giving over 75% peptide coverage for identification. The solvent system presented permits the detection of all five possible phosphorylation moieties. Application of histidine and aspartic acid phosphorylation modifications to proteomic analyses will significantly advance the understanding of phosphorylation relay signaling in cellular regulation, including elucidation of the role of NM23-H1 in metastasis.

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Section: Resultsmentioning

confidence: 99%

Mass Spectrometry Detection of Histidine Phosphorylation on NM23-H1

2010

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Section: Discussionmentioning

confidence: 97%

“…While this compound appears to impact invasiveness for some cancer cells by altering the DNA methylation status of a CpG island in the Nm23-H1 promoter, this is not the case in other cancer cells (23), suggesting that the effect of the compound may be cell type specific. Several other compounds in clinical use also upregulate Nm23-H1 expression in vitro and in vivo (23). Therefore, our data provide a rationale for pursuing additional studies to determine whether pharmacologic interventions resulting in cytoplasmic overexpression of Nm23-H1, or which target Nm23-H1-associated signal transduction, interfere with KSHV-associated invasiveness.…”

Section: Discussionmentioning

confidence: 97%

Regulation of Nm23-H1 and Cell Invasiveness by Kaposi's Sarcoma-Associated Herpesvirus

Qin

Dai

Toole

et al. 2011

J Virol

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The Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of Kaposi's sarcoma (KS), and the induction of an invasive cellular phenotype by KSHV following de novo infection is an important pathogenic component mediating tumor progression. The metastasis suppressor gene known as Nm23-H1 regulates tumor cell invasiveness, but whether KSHV itself regulates Nm23-H1 expression or subcellular localization, and whether this impacts cell invasiveness, has not been established. We found that KSHV increases expression and nuclear translocation of Nm23-H1 and that nuclear translocation of Nm23-H1 is regulated by the KSHV-encoded latency-associated nuclear antigen (LANA). Moreover, activation of the Ras-BRaf-MAPK (mitogen-activated protein kinase) signal transduction pathway, secretion of promigratory factors associated with this pathway, and cell invasiveness are dependent on KSHV regulation of Nm23-H1. Finally, induction of cytoplasmic overexpression of Nm23-H1 using a pharmacologic inhibitor of DNA methylation reduced KSHVassociated Ras-BRaf-MAPK pathway activation and suppressed KSHV-induced invasiveness. These data provide the first evidence for KSHV regulation of Nm23-H1 as a mechanism for KSHV induction of an invasive cellular phenotype and support the potential utility of targeting Nm23-H1 as a therapeutic approach for the treatment of KS.The Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi's sarcoma (KS) (8). Despite the reduced incidence of HIV-associated KS in the era of highly active antiretroviral therapy (HAART) (49), KS still represents one of the most common HIV-associated tumors in the modern era (6, 11) and an important cause of morbidity and mortality in patients receiving solid organ transplants (21). Acquisition of a migratory or invasive phenotype represents one hallmark of KSHV-infected cells and an important pathogenic mechanism for KS formation (29). A better understanding of KSHV-host cell interactions regulating cell invasiveness may help to identify mechanisms for KS pathogenesis and novel therapeutic targets.Acquisition of an invasive phenotype is negatively regulated by metastasis suppressor genes (MSGs), and over 20 MSGs have been identified to date (17, 34). The first well-characterized MSG, a histidine kinase known as Nm23-H1 (41), inhibits metastasis through cooperative mechanisms, including the phosphorylation of key intracellular proteins regulating signal transduction and the alteration of gene expression (42). Published data suggest that altered Nm23-H1 expression correlates with tumor progression and outcomes (7,13,16,22,26,35,48). However, these data are conflicting, with some studies suggesting that reduced Nm23-H1 expression leads to tumor progression and reduced patient survival (7,16,22,35), while others suggest that increased Nm23-H1 expression portends reduced survival (13,26,48). Therefore, Nm23-H1 regulation of pathogenesis may be tumor specific, and identification of regulatory mechanisms for Nm23-H1 expression and cellular locali...

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“…By contrast, histidine kinases are well known in bacteria and lower eukaryotes, in which they form a major signal transduction pathway to extracellular events (two-component-signaltransduction systems) [82,83]. In model studies with human breast carcinoma cells, it could be shown that the metastasis potential was decreased upon treatment with compounds that were capable to increase NDKA expression [84]. Recently, pharmaceutical companies have identified histidine kinase inhibitors as a next generation approach to antibioticresistant bacterial infection [85], and these technologies and agents may be readily transferable to the cancer field, once anti-metastatic activity is proven.…”

Section: Discussionmentioning

confidence: 99%

Mass Spectrometric Characterization of Protein Structure Details Refines the Proteome Signature for Invasive Ductal Breast Carcinoma

Röwer

Koy

Hecker

et al. 2011

J. Am. Soc. Mass Spectrom.

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Early diagnosis as well as individualized therapies are necessary to reduce the mortality of breast cancer, and personalized patient care strategies rely on novel prognostic or predictive factors. In this study, with six breast cancer patients, 2D gel analysis was applied for studying protein expression differences in order to distinguish invasive ductal breast carcinoma, the most frequent breast tumor subtype, from control samples. In total, 1203 protein spots were assembled in a 2D reference gel. Differentially abundant spots were subjected to peptide mass fingerprinting for protein identification. Twenty proteins with their corresponding 38 differentially expressed 2D gel spots were contained in our previously reported proteome signature, suggesting that distinct protein forms were contributing. In-depth MS/MS measurements enabled analyses of protein structure details of selected proteins. In protein spots that significantly contributed to our signature, we found that glyceraldehyde-3-phosphate dehydrogenase was N-terminally truncated, pyruvate kinase M2 and nucleoside diphosphate kinase A but not other isoforms of these proteins were of importance, and nucleophosmin phosphorylation at serine residues 106 and 125 were clearly identified. Principle component analysis and hierarchical clustering with normalized quantitative data from the 38 spots resulted in accurate separation of tumor from control samples. Thus, separation of tissue samples as in our initial proteome signature could be confirmed even with a different proteome analysis platform. In addition, detailed protein structure investigations enabled refining our proteome signature for invasive ductal breast carcinoma, opening the way to structure-/function studies with respect to disease processes and/or therapeutic intervention.

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Clinical-translational strategies for the elevation of Nm23-H1 metastasis suppressor gene expression

Cited by 17 publications

References 33 publications

Mass Spectrometry Detection of Histidine Phosphorylation on NM23-H1

Mass Spectrometry Detection of Histidine Phosphorylation on NM23-H1

Regulation of Nm23-H1 and Cell Invasiveness by Kaposi's Sarcoma-Associated Herpesvirus

Mass Spectrometric Characterization of Protein Structure Details Refines the Proteome Signature for Invasive Ductal Breast Carcinoma

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