The overall goal of the K08 award is to permit Laura J. Rush, D.V.M., to devote full-time effort for research training leading to a Ph.D. degree in molecular biology, and for development into an independent scientist for a career in academic biomedical research. Dr. Rush has completed a residency in veterinary and comparative pathology and 3 years of research in the laboratories of Michael A. Caligiuri, M.D., and Christoph Plass, Ph.D., co-sponsors of this award. Acute myeloid leukemia (AML) is a heterogeneous disease and the pathogenesis of most types of AML is unknown. The overall hypothesis of this research project is that aberrant DNA methylation in AML is associated with inactivation of genes that are necessary for normal growth, differentiation and/or death of hematopoietic cells, and t h at these epigenetic alterations play a key role in the molecular pathogenesis of AML. The proposed project will focus on fifteen candidate genes that have been identified by genome scanning experiments performed by Dr. Rush using primary AML samples. The goals of the project are to use these 15 genes to: 1) determine the consequences of restoration of gene expression on morphology, differentiation, and growth characteristics in vitro using AML cell lines; 2) investigate the methylation status of the 5' CpG islands by molecular techniques such as bisulfite treatment and methylation-specific in situ hybridization; and 3) correlate aberrant promoter methylation with transcriptional inactivation in AML diagnostic samples. Genes will be prioritized based on expression in normal hematopoietic tissues. The genes will then be transfected into non-expressing AML cell lines to elucidate what effects restoration of expression may have on growth rate, differentiation, b l a s t - colony forming ability, and morphology (Aim 1). Extensive characterization of the methylated 5' regulatory regions of these genes will be performed and results correlated with the presence or absence of the protein product (Aim 2). Dr. Rush has already developed extensive preliminary data for this project. The cooperative efforts of Dr. Rush, the co-sponsors, and co-investigators in the Division of Human Cancer Genetics, Comprehensive Cancer Center, and the Cancer and Leukemia Group B Tissue Bank will provide a productive environment to complete this significant investigation. The combination of results from studies of primary tumors, in vitro cultures, and mechanistic molecular experiments will contribute to the understanding of the role of aberrant DNA methylation in leukemogenesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08CA089317-03
Application #
6772640
Study Section
Subcommittee G - Education (NCI)
Program Officer
Eckstein, David J
Project Start
2002-07-01
Project End
2007-06-30
Budget Start
2004-07-01
Budget End
2005-06-30
Support Year
3
Fiscal Year
2004
Total Cost
$115,111
Indirect Cost
Name
Ohio State University
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
832127323
City
Columbus
State
OH
Country
United States
Zip Code
43210
Whitman, Susan P; Hackanson, Bjorn; Liyanarachchi, Sandya et al. (2008) DNA hypermethylation and epigenetic silencing of the tumor suppressor gene, SLC5A8, in acute myeloid leukemia with the MLL partial tandem duplication. Blood 112:2013-6
Smiraglia, Dominic J; Kazhiyur-Mannar, Ramakrishnan; Oakes, Christopher C et al. (2007) Restriction landmark genomic scanning (RLGS) spot identification by second generation virtual RLGS in multiple genomes with multiple enzyme combinations. BMC Genomics 8:446
Dorrance, Adrienne M; Liu, Shujun; Yuan, Weifeng et al. (2006) Mll partial tandem duplication induces aberrant Hox expression in vivo via specific epigenetic alterations. J Clin Invest 116:2707-16
Whitman, Susan P; Liu, Shujun; Vukosavljevic, Tamara et al. (2005) The MLL partial tandem duplication: evidence for recessive gain-of-function in acute myeloid leukemia identifies a novel patient subgroup for molecular-targeted therapy. Blood 106:345-52
Rush, Laura J; Raval, Aparna; Funchain, Pauline et al. (2004) Epigenetic profiling in chronic lymphocytic leukemia reveals novel methylation targets. Cancer Res 64:2424-33
Dai, Zunyan; Zhu, Wei-Guo; Morrison, Carl D et al. (2003) A comprehensive search for DNA amplification in lung cancer identifies inhibitors of apoptosis cIAP1 and cIAP2 as candidate oncogenes. Hum Mol Genet 12:791-801
Dai, Zunyan; Weichenhan, Dieter; Wu, Yue-Zhong et al. (2002) An AscI boundary library for the studies of genetic and epigenetic alterations in CpG islands. Genome Res 12:1591-8
Rush, Laura J; Plass, Christoph (2002) Alterations of DNA methylation in hematologic malignancies. Cancer Lett 185:1-12
Rush, Laura J; Plass, Christoph (2002) Restriction landmark genomic scanning for DNA methylation in cancer: past, present, and future applications. Anal Biochem 307:191-201