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153. A New Model of Epstein-Barr Virus Infection Reveals an Important Role for Early Lytic Viral Protein Expression in the Development of Lymphomas

Author

Ma, Shi-Dong, primary, Hegde, Subramanya, additional, Young, Ken H., additional, Sullivan, Ruth, additional, Rajesh, Deepika, additional, Zhou, Ying, additional, Jankowska-Gan, Ewa, additional, Burlingham, William J., additional, Sun, Xiaoping, additional, Gulley, Margaret L., additional, Tang, Weihua, additional, Gumperz, Jenny E., additional, and Kenney, Shannon C., additional

Published
2011
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156. Viral Response to Chemotherapy in Endemic Burkitt Lymphoma

Author

Tang, Weihua, primary, Harmon, Paula, additional, Gulley, Margaret L., additional, Mwansambo, Charles, additional, Kazembe, Peter N., additional, Martinson, Francis, additional, Wokocha, Clifford, additional, Kenney, Shannon C., additional, Hoffman, Irving, additional, Sigel, Carlie, additional, Maygarden, Susan, additional, Hoffman, Mariah, additional, and Shores, Carol, additional

Published
2010
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167. Distribution of Epstein‐Barr viral load in serum of individuals from nasopharyngeal carcinoma high‐risk families in Taiwan

Author

Yang, Xiaohong (Rose), primary, Goldstein, Alisa M., additional, Chen, Chien‐Jen, additional, Rabkin, Charles S., additional, Chen, Jen‐Yang, additional, Cheng, Yu‐Juen, additional, Hsu, Wan‐Lun, additional, Sun, Brenda, additional, Diehl, Scott R., additional, Liu, Mei‐Ying, additional, Walters, Michael, additional, Shao, Wen, additional, Ortiz‐Conde, Betty A., additional, Whitby, Denise, additional, Elmore, Sandra H., additional, Gulley, Margaret L., additional, and Hildesheim, Allan, additional

Published
2005
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171. Epstein–Barr viral load as a marker of lymphoma in AIDS patients

Author

Fan, Hongxin, primary, Kim, Seong Cheol, additional, Chima, Chukwuemeka O., additional, Israel, Bruce F., additional, Lawless, Kathleen M., additional, Eagan, Phyllis A., additional, Elmore, Sandra, additional, Moore, Dominic T., additional, Schichman, Steven A., additional, Swinnen, Lode J., additional, and Gulley, Margaret L., additional

Published
2004
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185. Comparative analysis of the expression of the epstein‐barr virus (EBV) anti‐apoptotic gene BHRF1 in nasopharyngeal carcinoma and EBV‐related lymphoid diseases

Author

Nicholls, John, primary, Kremmer, Elisabeth, additional, Meseda, Clement A., additional, Mackett, Mike, additional, Hahn, Peter, additional, Gulley, Margaret L., additional, Brink, Antoinette, additional, Swinnen, Lode J., additional, Greenspan, John, additional, De Souza, Yvonne, additional, Grässer, Friedrich, additional, Sham, Jonathan, additional, Ng, Mun‐Hon, additional, and Arrand, John R., additional

Published
2001
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186. Hepatosplenic ɑß T-Cell Lymphomas

Author

Macon, William R., primary, Levy, Norman B., additional, Kurtin, Paul J., additional, Salhany, Kevin E., additional, Elkhalifa, Mohamed Y., additional, Casey, Terence T., additional, Craig, Fiona E., additional, Vnencak-Jones, Cindy L., additional, Gulley, Margaret L., additional, Park, Jonathan P., additional, and Cousar, John B., additional

Published
2001
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188. Molecular Methods for Detecting Epstein-Barr Virus (Part III).

Author

Walker, John M., Killeen, Anthony A., Fan, Hongxin, and Gulley, Margaret L.

Abstract

Epstein-Barr virus (EBV) viral load testing is rapidly gaining acceptance in the diagnosis and monitoring of patients with EBV-related neoplasia, including transplant recipients, autoimmune deficiency syndrome (AIDS) patients with brain lymphoma, and patients with nasopharyngeal carcinoma. In transplant recipients, several studies have shown that EBV viral load in blood is a useful marker of EBV-driven posttransplant lymphoproliferative disease (PTLD), both in terms of predicting disease and in monitoring efficacy of therapy (1-4). In AIDS patients, high EBV viral load in cerebrospinal fluid (CSF) is so characteristic of brain lymphoma that the assay has been touted as sufficient for making that diagnosis without the need for brain biopsy, assuming that there is also clinical and radiographic support for the diagnosis (5). In nasopharyngeal carcinoma patients, a recent study showed that plasma EBV viral load is nearly always elevated, and the degree of elevation is higher in those with distant metastases (6). Additional studies are needed to more completely define the utility of EBV viral load measurement in monitoring residual disease following therapy, and to evaluate clinical utility in other EB V-associated diseases. [ABSTRACT FROM AUTHOR]

Published
2001
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189. RNA Extraction from Fresh or Frozen Tissues.

Author

Walker, John M., Killeen, Anthony A., Fan, Hongxin, and Gulley, Margaret L.

Abstract

RNA can be isolated from fresh or frozen tissue, then purified and quantified for subsequent molecular analysis. RNA is quite labile compared to DNA for good reason: RNA is the transient message that transmits information from activated genes. Once gene transcription is turned off, no more RNA is produced, and preexisting RNA is rapidly degraded to prevent continued translation of proteins. Because RNA is so labile, special care is required at all steps of RNA extraction to prevent RNA degradation. [ABSTRACT FROM AUTHOR]

Published
2001
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190. DNA Extraction from Fresh or Frozen Tissues.

Author

Walker, John M., Killeen, Anthony A., Fan, Hongxin, and Gulley, Margaret L.

Abstract

The first step in molecular analysis of patient tissues is preparation of purified, high molecular weight DNA. A number of methods and commercial kits are available for DNA isolation. Traditional organic extraction protocols (1,2) are based on the fact that DNA is soluble in water whereas lipids are soluble in phenol. In these protocols, tissues are disaggregated and then treated with detergent to lyse cell membranes followed by proteinase to digest proteins. Phenol, an organic solvent, is added to help separate the lipids and protein remnants from the DNA. Chloroform is then used to facilitate the removal of phenol. DNA is subsequently concentrated and further purified by precipitation in a cold mixture of salt and ethanol. Finally, DNA is resolubilized in Tris-EDTA buffer. [ABSTRACT FROM AUTHOR]

Published
2001
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191. DNA Extraction from Paraffin-Embedded Tissues.

Author

Walker, John M., Killeen, Anthony A., Fan, Hongxin, and Gulley, Margaret L.

Abstract

In routine histopathology, most tissues are fixed in formalin and embedded in paraffin for long-term preservation. DNA can be extracted from these tissues for subsequent molecular analysis by amplification methods. We describe herein a protocol for DNA preparation from paraffin-embedded tissues based on published procedures (1-3). In brief, tissue sections are placed into microfuge tubes, then deparaffinized with xylene. The xylene is removed with ethanol washes, and the tissue is treated with proteinase K to make DNA available for amplification. [ABSTRACT FROM AUTHOR]

Published
2001
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192. Gastric adenocarcinoma microRNA profiles in fixed tissue and in plasma reveal cancer-associated and Epstein-Barr virus-related expression patterns

Author

Treece, Amanda L, Duncan, Daniel L, Tang, Weihua, Elmore, Sandra, Morgan, Douglas R, Dominguez, Ricardo L, Speck, Olga, Meyers, Michael O, and Gulley, Margaret L

Abstract

MicroRNA expression in formalin-fixed paraffin-embedded tissue (FFPE) or plasma may add value for cancer management. The GastroGenus miR Panel was developed to measure 55 cancer-specific human microRNAs, Epstein-Barr virus (EBV)-encoded microRNAs, and controls. This Q-rtPCR panel was applied to 100 FFPEs enriched for adenocarcinoma or adjacent non-malignant mucosa, and to plasma of 31 patients. In FFPE, microRNAs upregulated in malignant versus adjacent benign gastric mucosa were hsa-miR-21, -155, -196a, -196b, -185, and -let-7i. Hsa-miR-18a, 34a, 187, -200a, -423-3p, -484, and -744 were downregulated. Plasma of cancer versus non-cancer controls had upregulated hsa-miR-23a, -103, and -221 and downregulated hsa-miR-378, -346, -486-5p, -200b, -196a, -141, and -484. EBV-infected versus uninfected cancers expressed multiple EBV-encoded microRNAs, and concomitant dysregulation of four human microRNAs suggests that viral infection may alter cellular biochemical pathways. Human microRNAs were dysregulated between malignant and benign gastric mucosa and between plasma of cancer patients and non-cancer controls. Strong association of EBV microRNA expression with known EBV status underscores the ability of microRNA technology to reflect disease biology. Expression of viral microRNAs in concert with unique human microRNAs provides novel insights into viral oncogenesis and reinforces the potential for microRNA profiles to aid in classifying gastric cancer subtypes. Pilot studies of plasma suggest the potential for a noninvasive addition to cancer diagnostics.

Published
2016
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196. Validation of the VE1 immunostain for the BRAF V600E mutation in melanoma.

Author

Pearlstein, Michelle V., Zedek, Daniel C., Ollila, David W., Treece, Amanda, Gulley, Margaret L., Groben, Pamela A., and Thomas, Nancy E.

Subjects
IMMUNOSTAINING, NEUROENDOCRINE tumors, MELANOMA, GENETIC mutation, GENETICS
Abstract

Background BRAF mutation status, and therefore eligibility for BRAF inhibitors, is currently determined by sequencing methods. We assessed the validity of VE1, a monoclonal antibody against the BRAF V600E mutant protein, in the detection of mutant BRAF V600E melanomas as classified by DNA pyrosequencing. Methods The cases were 76 metastatic melanoma patients with only one known primary melanoma who had had BRAF codon 600 pyrosequencing of either their primary (n = 19), metastatic (n = 57) melanoma, or both (n = 17). All melanomas (n = 93) were immunostained with the BRAF VE1 antibody using a red detection system. The staining intensity of these specimens was scored from 0 to 3+ by a dermatopathologist. Scores of 0 and 1+ were considered as negative staining while scores of 2+ and 3+ were considered positive. Results The VE1 antibody showed a sensitivity of 85% and a specificity of 100% as compared to DNA pyrosequencing results. There was 100% concordance between VE1 immunostaining of primary and metastatic melanomas from the same patient. V600K, V600Q, and V600R BRAF melanomas did not positively stain with VE1. Conclusions This hospital-based study finds high sensitivity and specificity for the BRAF VE1 immunostain in comparison to pyrosequencing in detection of BRAF V600E in melanomas. [ABSTRACT FROM AUTHOR]

Published
2014
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197. Case-case comparison of smoking and alcohol risk associations with Epstein-Barr virus-positive gastric cancer.

Author

Camargo, M. Constanza, Koriyama, Chihaya, Matsuo, Keitaro, Kim, Woo‐Ho, Herrera‐Goepfert, Roberto, Liao, Linda M., Yu, Jun, Carrasquilla, Gabriel, Sung, Joseph J.Y., Alvarado‐Cabrero, Isabel, Lissowska, Jolanta, Meneses‐Gonzalez, Fernando, Yatabe, Yashushi, Ding, Ti, Hu, Nan, Taylor, Philip R., Morgan, Douglas R., Gulley, Margaret L., Torres, Javier, and Akiba, Suminori

Abstract

Helicobacter pylori is the primary cause of gastric cancer. However, monoclonal Epstein-Barr virus (EBV) nucleic acid is also present in up to 10% of these tumors worldwide. EBV prevalence is increased with male sex, nonantral localization and surgically disrupted anatomy. To further examine associations between EBV and gastric cancer, we organized an international consortium of 11 studies with tumor EBV status assessed by in situ hybridization. We pooled individual-level data on 2,648 gastric cancer patients, including 184 (7%) with EBV-positive cancers; all studies had information on cigarette use (64% smokers) and nine had data on alcohol (57% drinkers). We compared patients with EBV-positive and EBV-negative tumors to evaluate smoking and alcohol interactions with EBV status. To account for within-population clustering, multilevel logistic regression models were used to estimate interaction odds ratios (OR) adjusted for distributions of sex (72% male), age (mean 59 years), tumor histology (56% Lauren intestinal-type), anatomic subsite (61% noncardia) and year of diagnosis (1983-2012). In unadjusted analyses, the OR of EBV positivity with smoking was 2.2 [95% confidence interval (CI) 1.6-3.2]. The OR was attenuated to 1.5 (95% CI 1.01-2.3) by adjustment for the possible confounders. There was no significant interaction of EBV status with alcohol drinking (crude OR 1.4; adjusted OR 1.0). Our data indicate the smoking association with gastric cancer is stronger for EBV-positive than EBV-negative tumors. Conversely, the null association with alcohol does not vary by EBV status. Distinct epidemiologic characteristics of EBV-positive cancer further implicate the virus as a cofactor in gastric carcinogenesis. [ABSTRACT FROM AUTHOR]

Published
2014
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