Molecular Profiling of Tumor Progression in Head and Neck Cancer

Belbin, Thomas J.; Singh, Bhuvanesh; Smith, Richard V.; Socci, Nicholas D.; Wreesmann, Volkert B.; Sänchez‐Carbayo, Marta; Masterson, Jessica; Patel, Snehal G.; Cordon‐Cardo, Carlos; Prystowsky, Michael B.; Childs, Geoffrey

doi:10.1001/archotol.131.1.10

Cited by 82 publications

(73 citation statements)

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“…Linear amplification of total RNA and subsequent fluorescent labeling of corresponding cDNA with Cy5 and Cy3 dyes were carried out using the MessageAmp T7 linear amplification kit (Ambion), and cDNA labeling protocols were developed at the AECOM microarray facility (11). Red (Cy5) and green (Cy3) signal intensities for each element on the array were calculated using GenePix Pro 3.0 software (Axon Instruments) package as described previously (11). cDNA arrays consisted of 27,323 sequence-verified human cDNA clones, representing both known genes and expressed sequence tags.…”

Section: Methodsmentioning

confidence: 99%

“…Gene expression profiles focused on alterations caused by MTDIA treatment in A549 xenograft tumors, detected with cDNA microarrays of 27,323 clones (11). 125 mRNAs with the greatest changes, defined by a Ͼ3.0 or Ͻ0.33 expression ratios relative to untreated tumors (supplemental Table S3A), represented many different functional categories.…”

Section: Gene Expression In A549 Tumorsmentioning

confidence: 99%

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Growth and Metastases of Human Lung Cancer Are Inhibited in Mouse Xenografts by a Transition State Analogue of 5′-Methylthioadenosine Phosphorylase

Basu

Locker

Cassera

et al. 2011

Journal of Biological Chemistry

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110

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The S-adenosylmethionine (AdoMet) salvage enzyme 5 -methylthioadenosine phosphorylase (MTAP) has been implicated as both a cancer target and a tumor suppressor. We tested these hypotheses in mouse xenografts of human lung cancers. AdoMet recycling from 5 -methylthioadenosine (MTA) was blocked by inhibition of MTAP with methylthioDADMe-Immucillin-A (MTDIA), an orally available, nontoxic, picomolar transition state analogue. Blood, urine, and tumor levels of MTA increased in response to MTDIA treatment. Disruption of pathways linked to polyamine synthesis and S-adenosylmethionine (AdoMet) 2 salvage provides metabolic targets in anticancer therapy based on the essential roles of these metabolites in cell growth. AdoMet is the major methyl donor for biosynthetic methylation reactions, a precursor for polyamine synthesis, and the source of methyl groups for DNA methylation. Targeting polyamine metabolism directly at L-ornithine decarboxylase by ␣,␣-difluoromethylornithine has had limited anticancer success (1). Two AdoMet molecules are converted to 5Ј-methylthioadenosine (MTA) in spermine synthesis, and 5Ј-methylthioadenosine phosphorylase (MTAP) recycles MTA by phosphorolysis to permit subsequent resynthesis of AdoMet (Fig. 1). Our working hypothesis was that inhibition of MTAP would affect cellular MTA and AdoMet metabolism with downstream effects on protein, DNA methylation, polyamine synthesis, and polyamine-dependent enzyme reactions. We targeted MTAP by transition state analysis and developed methylthio-DADMe-Immucillin-A (MTDIA), an orally available transition state analogue inhibitor of MTAP (2). Others have proposed that MTAP is a tumor suppressor gene (3), and experiments here explore the effects of MTAP inhibition in human lung cancer xenografts.We previously demonstrated that treatment of human FaDu head and neck tumors in mouse xenografts with MT-DIA prevented tumor growth with no apparent toxicity to the mice (4). Here, we report that both MTAP-positive (H358) and MTAP-deleted (A549) human lung cancer cell lines are also sensitive to MTAP inhibition in mouse xenografts. The mechanism is probed by the metabolic and genetic consequences of MTDIA administration. In culture, MTDIA in combination with MTA slows A549 cell growth but induces apoptosis in H358 cells.Lung cancer is the leading cause of cancer-related deaths worldwide (5). Patients diagnosed at an advanced stage have a median survival of less than 12 months (6 -8). Thus, development of novel therapeutics for lung cancer is a research priority. MTDIA demonstrated significant suppression of tumor growth with human lung cancer A549 and H358 cells in mouse xenografts. Low toxicity, oral availability, and significant tumor suppression by MTDIA all support additional evaluation as an agent for the treatment of lung cancers. EXPERIMENTAL PROCEDURESCell Lines-Human non-small cell lung adenocarcinoma (NSCLC) cell line A549 and prostate carcinoma cell line PC3 were obtained from the American Type Culture Collection (Manassas, VA). Human bronchioloalveolar ...

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Section: Methodsmentioning

confidence: 99%

Section: Gene Expression In A549 Tumorsmentioning

confidence: 99%

Growth and Metastases of Human Lung Cancer Are Inhibited in Mouse Xenografts by a Transition State Analogue of 5′-Methylthioadenosine Phosphorylase

Basu

Locker

Cassera

et al. 2011

Journal of Biological Chemistry

Self Cite

110

View full text Add to dashboard Cite

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“…Linear amplification of primary tumor total RNA and subsequent fluorescent labeling of corresponding cDNA was carried out using the MessageAmp T7 linear amplification kit (Ambion) and cDNA labeling protocols developed at the AECOM Microarray Facility (http://microarray1k.aecom.yu.edu; [11]). In order to optimize differences in gene expression profiles among the individual primary tumors, we first created pools of total RNA for each anatomic site by pooling equal aliquots of total RNA from each of the 12 primary tumors representing that specific site.…”

Section: T7 Linear Amplification Fluorescent Labeling and Hybridizamentioning

confidence: 99%

“…This approach has historically been utilized in the identification of differentially expressed genes associated with carcinogenesis [11,14,15]. Drawing from the same patient population as for previous analyses, we selected 12 patients (6 OP cases, 6 LH cases) and utilized the same cDNA microarray technology to compare primary HNSCC tumor RNA (Cy3-green) to normal adjacent mucosa (Cy5-red) from the same patient.…”

Section: Differential Gene Expression Between Primary Hnscc Tumors Anmentioning

confidence: 99%

Site-Specific Molecular Signatures Predict Aggressive Disease in HNSCC

Belbin

Smith

Adrien

et al. 2008

Head and Neck Pathol

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It is known that head and neck squamous cell carcinomas (HNSCC) originating from different anatomic locations can exhibit varying behavior that is not predictable by histopathology of the primary tumor. Using a microarray containing 27,323 cDNA clones, we generated sets of gene expression profiles for 36 HNSCC primary tumors (12 oral cavity, 12 oropharynx, and 12 larynx/ hypopharynx). From these datasets, we ranked genes according to their ability to differentiate between patients whose disease progressed within a 24 month period (aggressive phenotype) and those that did not (nonaggressive phenotype) based on levels of gene expression. A merging of datasets from the three sites revealed that only a fraction of identified genes were shared between any two sites. This contrasted greatly with the significant overlap (approximately 50%) in down-regulated genes identified in tumor/normal comparisons using cases both from oropharynx and larynx/hypopharynx. From these data, we conclude that HNSCC tumors originating from different anatomic sites share consistent changes in gene expression when comparing primary tumors to normal adjacent mucosa; these common changes most likely reflect alterations required for tumor development. In contrast, once a tumor has developed, tumor-host interactions at the different anatomic sites are likely responsible for the site-specific signatures associated with aggressive versus non-aggressive disease. Predictions of outcome based on gene expression profiling are therefore heavily influenced by the anatomic site of the primary tumor.

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“…Various studies have revealed numerous molecular abnormalities in HNSCC, including activation of oncogenes such as CCND1 (9), EGFR (10), RAS (11) and C-MYC (12); inactivation of tumor suppressor genes such as CDKN2A (p16) (13), TP53 (14), p27 (15) and WAF1/C1P1 (16); expression of angiogenic factors and LOH at numerous chromosomal locations (17,18). For instance, the most frequent cytogenetic alterations in HNSCC are gains of 3q, 8q, 20q, 7q, 11q13 and 5p, and losses in 3p, 9p, 21q, 5q, 13q, 18q and 8p (19). The involved loci correspond to genes that encode specific functional classes of proteins, such as cell cycle regulators, tumor suppressors, cell adhesion and protein kinases (20).…”

Section: Introductionmentioning

confidence: 99%