The long term goal of this project is to define the genetic changes associated with AML relapse/resistance to chemotherapy. This work requires paired samples of de novo and relapsed AML cells that are nearly homogeneous in composition, and tissue culture and mouse models to validate the functional consequences of these genetic changes. The Genomics of AML PPG provides the appropriate infrastructure for this project. We will investigate AML relapse/resistance via the following Specific Aims:
Specific Aim 1 : We will define the genetic changes that occur in murine APL cell lines selected for chemotherapeutic resistance in vitro. We have generated 10 "parental" murine APL tumor cell lines. We will generate a total of 10 murine APL tumor cell lines and paired subclones that are resistant to daunorubicin (DNR), and/or Ara-C both in vitro and in vivo. Using these well-defined clonal populations of cells, we will perform gene expression profiling and we will define acquired microdeletions and amplifications using array-based comparative genomic hybridization (CGH) with the NimbleGen 2.1M murine oligomer array. Genes that are consistently dysregulated, deleted, or amplified in DNR or Ara-C resistant subclones will be validated with qPCR approaches. Selected genes identified with these array-based genomic screens will be resequenced to define more subtle genetic changes. Functional validation will be performed using forced overexpression and shRNAi knock-down approaches.
Specific Aim 2 : We will define the genetic changes that contribute to AML relapse by comparing the genomes of AML cells obtained at initial presentation vs. first relapse. Because most relapsed samples are not well matched to the paired de novo samples in terms of cellular composition, we will purify AML blasts by sorting "blast gate" AML cells from the cte novo and relapsed sample pairs for at least 20 AML patients. Using RNA and DMA from these paired, enriched samples, we will perform array based expression profiling and high resolution array based CGH using the 2.1M human oligomer arrays from NimbleGen. The altered genes identified in Aims 1 and 2 will be used to select a subset of target genes for resequencing and biologic validation in the mouse APL model described in Aim 1.
Specific Aim 3 : We will assess the role of the bone marrow microenvironment on AML resistance and relapse. We will use a unique mouse model in which genetically-marked murine APL cells home to and expand in the mouse bone marrow (BM). We will determine whether interruption of the protective AML cell-stromal interaction (using inhibitors of the SDF-1-CXCR4 and the VCAM-1-VLA-4 axes) can sensitize APL cells to chemotherapy in vivo. Finally, we have devised a clinical trial in which we will test the role of a small molecule inhibitor of the CXCR4-SDF-1 axis given immediately prior to salvage chemotherapy in patients with relapsed AML to enhance remission rates and overall survival.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Program Projects (P01)
Project #
Application #
Study Section
Special Emphasis Panel (ZCA1-GRB-S)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Washington University
Saint Louis
United States
Zip Code
Al-Hussaini, Muneera; Rettig, Michael P; Ritchey, Julie K et al. (2016) Targeting CD123 in acute myeloid leukemia using a T-cell-directed dual-affinity retargeting platform. Blood 127:122-31
Welch, John S; Petti, Allegra A; Miller, Christopher A et al. (2016) TP53 and Decitabine in Acute Myeloid Leukemia and Myelodysplastic Syndromes. N Engl J Med 375:2023-2036
Wong, Terrence N; Miller, Christopher A; Klco, Jeffery M et al. (2016) Rapid expansion of preexisting nonleukemic hematopoietic clones frequently follows induction therapy for de novo AML. Blood 127:893-7
Griffith, Malachi; Griffith, Obi L; Krysiak, Kilannin et al. (2016) Comprehensive genomic analysis reveals FLT3 activation and a therapeutic strategy for a patient with relapsed adult B-lymphoblastic leukemia. Exp Hematol 44:603-13
Cole, Christopher B; Verdoni, Angela M; Ketkar, Shamika et al. (2016) PML-RARA requires DNA methyltransferase 3A to initiate acute promyelocytic leukemia. J Clin Invest 126:85-98
Churpek, Jane E; Pyrtel, Khateriaa; Kanchi, Krishna-Latha et al. (2015) Genomic analysis of germ line and somatic variants in familial myelodysplasia/acute myeloid leukemia. Blood 126:2484-90
Wong, Terrence N; Ramsingh, Giridharan; Young, Andrew L et al. (2015) Role of TP53 mutations in the origin and evolution of therapy-related acute myeloid leukaemia. Nature 518:552-5
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
Griffith, Malachi; Miller, Christopher A; Griffith, Obi L et al. (2015) Optimizing cancer genome sequencing and analysis. Cell Syst 1:210-223
Lu, Charles; Xie, Mingchao; Wendl, Michael C et al. (2015) Patterns and functional implications of rare germline variants across 12 cancer types. Nat Commun 6:10086

Showing the most recent 10 out of 98 publications