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Research Description:
My research group is part of--and located within--a larger scientific entity, the laboratory of Dr. Nikolaus Trede at the Huntsman Cancer Institute. The Trede group uses the zebrafish (Danio rerio) as a model organism for studies into several aspects of the vertebrate immune system. Specifically, efforts are focused upon thymic and T lymphocyte development, T cell-derived malignancies, macrophage development, and translational investigations into T cell-specific cytotoxic compounds. Within this context, I am conducting two research projects:
(1) Changes to genome-wide DNA methylation and transcription that accompany lymphocyte commitment to the B and T cell lineages. Cell populations being studied thus far include immature and mature B cells, immature T lymphocytes (thymocytes), and mature T cells. Lymphocyte populations from adult fish are sub-fractionated using fluorescence-activated cell sorting (FACS) methodologies. Nucleic acids (genomic DNA, total RNA) are then purified. To enrich the methylated DNA portion of the genome, immunoprecipitation using an anti-5'-methylcytosine antibody is performed. Finally, immunoprecipitated methyl-DNA and total RNA pools are then hybridized to microarrays containing representations of the entire zebrafish genome. Using this strategy, we are attempting to discover genes with key roles governing the choice in fate between T versus B lineage, as well as immature vs. mature lymphocyte.
(2) Forward genetic identification of mutant zebrafish lines with a predisposition to T cell malignancies. Using a forward-genetic screening approach of chemically-mutagenized zebrafish which harbor fluorescent T lymphocytes, we have discovered several distinct genetic lines with a predilection to T cell leukemias and lymphomas. Each line is being investigated in several ways: (1) phenotypic characterization of their respective malignancies, via immunohistochemical and gene-expression approaches, (2) establishment of conditions for in vitro culture of malignant cells, (3) mapping of the mutated genes underlying these different lines' malignant predispositions, and (4) genome-wide DNA methylation and transcription studies to distinguish the methylation and transcription 'signatures' that define these malignant cells' fates.
Research Keywords:
Zebrafish, T lymphocyte, leukemia, lymphoma, DNA Methylation, T cell development