The long-term goal of this project is to determine whether the clonal architecture of AML samples is relevant for clinical outcomes, and whether that information can be translated into clinical testing to predict prognosis. Our group has recently discovered that the clonal architecture of AML samples can be deduced by whole genome sequencing (which defines ali mutations in every case), followed by deep digital sequencing of ail mutations to define their variant allele frequencies (VAFs). Groups of mutations with similar VAFs represent subclones, Ali AML samples have a founding clone, and most contain 1-3 additional subclones that are derived from the founding clone. The dominant clone at relapse, however, is often a subclone that has acquired addition mutations. These data suggest that specific subclones contain mutations that are relevant for altered grov/th properties and/or altered drug sensitivity, which may contribute to refractory disease or relapse. In this project, we will study the clonal architecture of AML genomes, and determine their relevance for ciinical outcomes, via the following specific Aims:
Specific Aim 1 : We will use custom capture reagents and deep digital sequencing to determine the rate of clearance of individual AML subclones after induction chemotherapy. We will utilize DNA derived from bone marrow biopsies of 89 AML samples that have already undergone whole genome sequencing. gDNA from bone marrow biopsies obtained 2-4 weeks after initiation of therapy will be subjected to custom capture for all known mutations in each genome, and deep digital sequencing will be performed to assess the rate of clearance of the founding clone and all subclones. Clearance patterns will be correlated with clinical parameters to define impact on outcomes.
Specific Aim 2 : We will use a stromal-based culture system and xenotransplantation to evaluate the clonal architecture of AML samples grown in vitro and in vivo. Our stromal-based culture system allows for the expansion of most primary AML samples for at least 7 days without significantly altered physical properties or clonal drift. We will treat cultured AML cells with a variety of drugs, and responses will be measured using cell cycle assays and deep digital sequencing to define the drug sensitivity of each sample and each subclone. We will also analyze AML cells that expand in immunodeficient mice to determine whether specific subclones have a growth advantage in mice. These data will be used to identify mutations in subclones that may negatively influence prognosis.

Public Health Relevance

A better understanding ofthe clonal heterogeneity of Acute Myeloid Leukemia is required to understand why the disease is sometimes resistant to standard therapies. Genomic studies, coupled with new methods to expand human AML cells in vitro and in vivo, may provide new approaches for identifying high-risk patients at presentation, and may suggest new therapeutic approaches for these patients.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA101937-11
Application #
8696959
Study Section
Special Emphasis Panel (ZCA1-RPRB-J)
Project Start
Project End
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
11
Fiscal Year
2014
Total Cost
$320,105
Indirect Cost
$101,872
Name
Washington University
Department
Type
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Engle, E K; Fisher, D A C; Miller, C A et al. (2015) Clonal evolution revealed by whole genome sequencing in a case of primary myelofibrosis transformed to secondary acute myeloid leukemia. Leukemia 29:869-76
Al-Hussaini, Muneera; DiPersio, John F (2014) Small molecule inhibitors in acute myeloid leukemia: from the bench to the clinic. Expert Rev Hematol 7:439-64
Jacoby, M A; De Jesus Pizarro, R E; Shao, J et al. (2014) The DNA double-strand break response is abnormal in myeloblasts from patients with therapy-related acute myeloid leukemia. Leukemia 28:1242-51
Sarkaria, S M; Christopher, M J; Klco, J M et al. (2014) Primary acute myeloid leukemia cells with IDH1 or IDH2 mutations respond to a DOT1L inhibitor in vitro. Leukemia 28:2403-6
Miller, Christopher A; White, Brian S; Dees, Nathan D et al. (2014) SciClone: inferring clonal architecture and tracking the spatial and temporal patterns of tumor evolution. PLoS Comput Biol 10:e1003665
Klco, Jeffery M; Spencer, David H; Miller, Christopher A et al. (2014) Functional heterogeneity of genetically defined subclones in acute myeloid leukemia. Cancer Cell 25:379-92
Russler-Germain, David A; Spencer, David H; Young, Margaret A et al. (2014) The R882H DNMT3A mutation associated with AML dominantly inhibits wild-type DNMT3A by blocking its ability to form active tetramers. Cancer Cell 25:442-54
Hughes, Andrew E O; Magrini, Vincent; Demeter, Ryan et al. (2014) Clonal architecture of secondary acute myeloid leukemia defined by single-cell sequencing. PLoS Genet 10:e1004462
White, Brian S; DiPersio, John F (2014) Genomic tools in acute myeloid leukemia: From the bench to the bedside. Cancer 120:1134-44
Grieselhuber, N R; Klco, J M; Verdoni, A M et al. (2013) Notch signaling in acute promyelocytic leukemia. Leukemia 27:1548-57

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