EXCEED THE SPACE PROVIDED. R21Phase Our preliminary data indicate that freely circulating methylated DNA can be found in the plasma of patients with esophageal cancer. Specifically, these data show that methylated alleles of the APC, HPP1 and pi6 genes can be detected in the bloodstream of esophageal cancer patients. Furthermore, our preliminary data show decreased survival in patients with high plasma levels of methylated APC DNA, which also parallel relapse of malignancy. Thus, these circulating methylated nucleic acids show promise as biomarkers for the prognostication and monitoring of esophageal cancer patients. The overall scope of this proposed project will be to move these circulating biomarkers from the laboratory into the clinic. In the initial R21 phase, precise assays of gene-specific methylated plasma DNA will be developed and refined using quantitativereal-time methylation-specific PCR, with the established methylation targets pi6 and APC serving as templates. In addition, during this exploratory R21 phase, additional novel methylation targets will be identified in neoplastic esophageal tissues and plasma. In the second, R33 phase, novel methylation targets identified in the R21 phase will be clinically validated on a larger, independent cohort by performing clinical correlations with tissue and plasma methylation levels.
Aim 1. To develop and validate accurate, robust, standardizable, and scalable quantitative real-time plasma DNA methylation assays using the established tissue and plasma methylation targets p!6 and APC. Assays will be analytically validated with known standards of plasma from normal subjects spiked with known levels of genomic DNA, as well as known but blinded positive and negative patient blood samples.
Aim 2. To identify and measure 20 novel methylation events in 50 primary esophageal carcinomas and 50 normal esophageal tissues. A panel of candidate genes with known cancer and outcome relevance and reported frequent methylation in gastrointestinal and other human tumors will be evaluated using real-time quantitativemethylation- specific PCR (MSP). Genes methylated in at least 20% of tumors, but in less than 5% of normal specimens, will be further pursued in Aim 3.
Aim 3. To study the same pilot group of 50 patients forplasma methylation of target genes identified in tissues during Aim 2. Genes methylated in the plasma of greater than 10% of these patients will constitute plasma targets for clinical validation during the R33 phase of the current proposal. PERFORMANCE SITE ========================================Section End===========================================

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
7R21CA106763-02
Application #
7318779
Study Section
Special Emphasis Panel (ZRG1-CBSS (01))
Program Officer
Thurin, Magdalena
Project Start
2005-04-01
Project End
2009-03-31
Budget Start
2006-11-15
Budget End
2009-03-31
Support Year
2
Fiscal Year
2005
Total Cost
$35,169
Indirect Cost
Name
Johns Hopkins University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Ito, Tetsuo; Sato, Fumiaki; Kan, Takatsugu et al. (2011) Polo-like kinase 1 regulates cell proliferation and is targeted by miR-593* in esophageal cancer. Int J Cancer 129:2134-46
Jin, Zhe; Cheng, Yulan; Gu, Wen et al. (2009) A multicenter, double-blinded validation study of methylation biomarkers for progression prediction in Barrett's esophagus. Cancer Res 69:4112-5
Kan, Takatsugu; Sato, Fumiaki; Ito, Tetsuo et al. (2009) The miR-106b-25 polycistron, activated by genomic amplification, functions as an oncogene by suppressing p21 and Bim. Gastroenterology 136:1689-700
Selaru, Florin M; Olaru, Alexandru V; Kan, Takatsugu et al. (2009) MicroRNA-21 is overexpressed in human cholangiocarcinoma and regulates programmed cell death 4 and tissue inhibitor of metalloproteinase 3. Hepatology 49:1595-601
Akagi, Tadayuki; Ito, Tetsuo; Kato, Motohiro et al. (2009) Chromosomal abnormalities and novel disease-related regions in progression from Barrett's esophagus to esophageal adenocarcinoma. Int J Cancer 125:2349-59
Abraham, John M; Cheng, Yulan; Hamilton, James P et al. (2008) Generation of small 32P-labeled peptides as a potential approach to colorectal cancer therapy. PLoS One 3:e2508
Sato, Fumiaki; Jin, Zhe; Schulmann, Karsten et al. (2008) Three-tiered risk stratification model to predict progression in Barrett's esophagus using epigenetic and clinical features. PLoS One 3:e1890
Ito, Tetsuo; Shimada, Yutaka; Kan, Takatsugu et al. (2008) Pituitary tumor-transforming 1 increases cell motility and promotes lymph node metastasis in esophageal squamous cell carcinoma. Cancer Res 68:3214-24
Jin, Zhe; Cheng, Yulan; Olaru, Alexandru et al. (2008) Promoter hypermethylation of CDH13 is a common, early event in human esophageal adenocarcinogenesis and correlates with clinical risk factors. Int J Cancer 123:2331-6
Jin, Zhe; Hamilton, James P; Yang, Jian et al. (2008) Hypermethylation of the AKAP12 promoter is a biomarker of Barrett's-associated esophageal neoplastic progression. Cancer Epidemiol Biomarkers Prev 17:111-7

Showing the most recent 10 out of 17 publications