The long-term objective of this research is to understand the influence of hematopoietic-specific developmental programs on the repair of one type of DNA damage--a double strand break (DSB)--and the initial molecular events that lead to translocations, which are a hallmark of leukemia, lymphoma, and soft-tissue sarcomas. Chromosomal DSBs are formed during normal metabolic processes including DNA replication and antigen receptor rearrangement in developing lymphoid cells, as well as following exposure to DNA damaging agents. There is an association between certain chemotherapeutic agents and secondary leukemia, e.g. treatment with topoisomerase II inhibitors and rearrangements involving 1lq23, and sequence analysis of breakpoints suggests that DNA damage is involved in the translocation process. However, the mechanisms by which specific translocations occur within developing hematopoietic subpopulations in the initial formation of rearrangements remain unclear. These questions can be addressed by adaptation of a genetic system developed in murine ES cells based on the rare-cutting yeast endonuclease I-Sce I to introduce DSBs at defined genomic loci and analyze recombinant repair events at the molecular level. This system also scores for translocations, duplications, and deletions that may result. This proposal will adapt the I-Sce I system to examine DSB repair and recombination in hematopoietic early progenitor and myeloid cell lineages and the potential for this type of damage to promote illegitimate recombination. Approaches will: (1) determine the potential for repair of DSBs and interchromosomal recombination to promote genome rearrangements within specific lineages; (2) determine the potential for repair of DSBs within the breakpoint cluster regions of the MLL and AF-4 genes to result in t(4;11) translocations similar to those observed in the clinical setting; and (3) use a genomics microarray-based approach to characterize the influence of the stage of hematopoietic development or the damaging agent on the specificity of a cell's DNA damage response. These studies will provide important insight into the biology of DSB rejoining in hematopoietic cell subpopulations, and the factors responsible for the normal suppression of genome rearrangements and, ultimately, tumorigenesis. Unraveling the etiology and consequences of translocations may lead to new approaches to therapy and prevention.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA100159-02
Application #
6749584
Study Section
Special Emphasis Panel (ZRG1-CAMP (01))
Program Officer
Pelroy, Richard
Project Start
2003-05-19
Project End
2007-04-30
Budget Start
2004-05-01
Budget End
2005-04-30
Support Year
2
Fiscal Year
2004
Total Cost
$286,862
Indirect Cost
Name
Columbia University (N.Y.)
Department
Miscellaneous
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
Bariar, Bhawana; Vestal, C Greer; Deem, Bradley et al. (2018) Bioflavonoids promote stable translocations between MLL-AF9 breakpoint cluster regions independent of normal chromosomal context: Model system to screen environmental risks. Environ Mol Mutagen :
Cupello, Steven; Richardson, Christine; Yan, Shan (2016) Cell-free Xenopus egg extracts for studying DNA damage response pathways. Int J Dev Biol 60:229-236
Richardson, Christine; Yan, Shan; Vestal, C Greer (2015) Oxidative stress, bone marrow failure, and genome instability in hematopoietic stem cells. Int J Mol Sci 16:2366-85
Bariar, Bhawana; Vestal, C Greer; Richardson, Christine (2013) Long-term effects of chromatin remodeling and DNA damage in stem cells induced by environmental and dietary agents. J Environ Pathol Toxicol Oncol 32:307-27
White, Ryan R; Sung, Patricia; Vestal, C Greer et al. (2013) Double-strand break repair by interchromosomal recombination: an in vivo repair mechanism utilized by multiple somatic tissues in mammals. PLoS One 8:e84379
Mouzannar, R; McCafferty, J; Benedetto, G et al. (2011) TRANSCRIPTIONAL AND PHOSPHO-PROTEOMIC SCREENS REVEAL STEM CELL ACTIVATION OF INSULIN-RESISTANCE AND TRANSFORMATION PATHWAYS FOLLOWING A SINGLE MINIMALLY TOXIC EPISODE OF ROS. Int J Genomics Proteomics 2:34-49
Pandita, Tej K; Richardson, Christine (2009) Chromatin remodeling finds its place in the DNA double-strand break response. Nucleic Acids Res 37:1363-77
Libura, Jolanta; Ward, Maureen; Solecka, Joanna et al. (2008) Etoposide-initiated MLL rearrangements detected at high frequency in human primitive hematopoietic stem cells with in vitro and in vivo long-term repopulating potential. Eur J Haematol 81:185-95
Koptyra, M; Cramer, K; Slupianek, A et al. (2008) BCR/ABL promotes accumulation of chromosomal aberrations induced by oxidative and genotoxic stress. Leukemia 22:1969-72
Francis, Richard; Richardson, Christine (2007) Multipotent hematopoietic cells susceptible to alternative double-strand break repair pathways that promote genome rearrangements. Genes Dev 21:1064-74

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