Project Details
Abstract
Backgrounds: Acute myeloid leukemia (AML) is a molecularly and clinically heterogeneous
myeloid neoplasm. AML with partial tandem duplication of MLL gene (MLL-PTD) is a specific
subtype and confers poor outcome. Our preliminary data showed a high frequency of RUNX1 or
DNMT3A mutations in de novo MLL-PTD AML patients and they were mutually exclusive
with rare exception. The cooperative biological function of MLL-PTD with RUNX1 mutations
or DNMT3A mutations is unclear. We hypothesize that MLL-PTD cooperating with RUNX1
mutations or DNMT3A mutations results in AML transformation and they might contribute to
leukemogenesis through the same or similar signaling pathways.
Aims: Our specific aims are: 1) to determine the cooperative function of MllPTD and RUNX1
mutants in initiating AML in mouse model, validate the expression of target genes in human
AML samples, and examine treatment strategies with specific inhibitors; 2) to determine the
cooperative biological function of MLL/MllPTD and DNMT3A mutants in the pathogenesis of
AML and explore the downstream targets as potential biomarkers and/or therapeutic targets.
Materials and Methods: After retroviral or lentiviral-mediated over-expression of RUNX1 or
DNMT3A mutants in MllPTD mouse bone marrow cells or human MLL-PTD positive AML cell
lines, we will perform colony formation assays, analyze cell growth/proliferation by MTS
assay and cellular differentiation by flow cytometry, and examine the dose responsive curve for
specific inhibitors. Microarray analysis will be performed to correlate gene expression and
DNA methylation status. We will identify differentially expressed genes in the transformed
cells or clinical samples harboring MLL-PTD in mRNA and/or protein levels by RT-qPCR
and/or Western blot. For in vivo studies, we will perform bone marrow transplantation assay for
promising RUNX1 or DNMT3A mutants in irradiated B6 mice to investigate leukemia
transformation and generate inducible double knock-in mice with cooperating MllPTD/WT and
Runx1Mut/WT driven by LSL-tTA/Vav1-Cre to check AML development. In the future, we
might use the double knock-in mouse model for drug treatment strategies.
Significance: After accomplishing this project, we will be able to better understand the
pathogenesis of de novo MLL-PTD AML and its cooperative roles in leukemogenesis with
RUNX1 or DNMT3A mutations, which in turn will provide an opportunity to discover new
biomarkers and identify novel therapeutic targets for the pre-clinical approaches
Project IDs
Project ID:PC10309-0064
External Project ID:MOST103-2321-B182-015
External Project ID:MOST103-2321-B182-015
Status | Finished |
---|---|
Effective start/end date | 01/08/14 → 31/07/15 |
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