Chemotherapy resistance is the most important clinical challenge in treating patients with acute myeloid leukemia (AML). The majority of patients with AML initially respond to chemotherapy but over half of responders will relapse and eventually die of disease. Traditionally, chemotherapy resistance is hypothesized to represent selection for a genetically distinct clone that has properties of leukemic stem cells (LSC). Recent data and our preliminary results (below) indicate that neither part of this model is fully correct. Rather, we propose that chemotherapy resistance can and frequently is an acquired epigenetic response to chemotherapeutic challenge that may arise in any leukemic clone. In vitro, we have modeled chemotherapy resistance and demonstrate that both DNA methylation and histone methylation are altered rapidly (within days) after treatment with Ara-C. Specifically, H3K27me3 is increased and H3K4me2 is decreased. In order to initially study the contribution of epigenetic changes in primary cells, we have compared the genetic and DNA methylation signature of 140 pairs of AML cells at diagnosis and relapse. Importantly, preliminary evidence suggests that at least 20% of patients have no new detectable genetic abnormalities at relapse. In contrast, when comparing paired diagnosis vs. relapse AML specimens we observe extensive and partially convergent redistribution of cytosine methylation affecting specific pathways, regardless of genetic background. Taken together these studies suggest an epigenetic mechanism of chemotherapy resistance but do not address the role of leukemic stem cells. To address this, we have studied the effect of cytosine arabinoside (Ara-C) on AML cells xenografted in NSG mice. Importantly, we demonstrate for the first time that limiting dilution analysis of Ara-C treatd AML cells does NOT show an enrichment in SCID-leukemia initiating cells. Collectively these data lead us to hypothesize that chemotherapy resistance in AML is a biologically complex event that includes but is not fully explained by genetic lesions and is composed of both fixed and inducible epigenetic elements. To test and explore these ideas, we propose 3 Specific Aims:
Specific Aim 1) Identify and characterize candidate mechanisms of epigenetic response to Ara-C in AML cells in vitro.
Specific Aim 2) Identify epigenetic and genetic determinants of relapse in primary AML patient samples.
Specific Aim 3) Determine the dynamics and mechanism through which chemoresistant features emerge in leukemic cell populations.

Public Health Relevance

Acute myeloid leukemia is a highly dangerous blood cancer with a high mortality rate. Most patients respond initially to chemotherapy but subsequently relapse with an incurable form of the illness. We propose a novel mechanism underlying chemotherapy resistance and describe approaches to reverse chemoresistance in order to develop better therapies for patients with AML.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA198089-05
Application #
9716381
Study Section
Basic Mechanisms of Cancer Therapeutics Study Section (BMCT)
Program Officer
Klauzinska, Malgorzata
Project Start
2015-07-22
Project End
2020-06-30
Budget Start
2019-07-01
Budget End
2020-06-30
Support Year
5
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Kunimoto, Hiroyoshi; Meydan, Cem; Nazir, Abbas et al. (2018) Cooperative Epigenetic Remodeling by TET2 Loss and NRAS Mutation Drives Myeloid Transformation and MEK Inhibitor Sensitivity. Cancer Cell 33:44-59.e8
Krevvata, Maria; Shan, Xiaochuan; Zhou, Chenghui et al. (2018) Cytokines increase engraftment of human acute myeloid leukemia cells in immunocompromised mice but not engraftment of human myelodysplastic syndrome cells. Haematologica 103:959-971
Bhavanasi, Dheeraj; Wen, Kwun Wah; Liu, Xiaolei et al. (2017) Signaling mechanisms that regulate ex vivo survival of human acute myeloid leukemia initiating cells. Blood Cancer J 7:636
Kunimoto, Hiroyoshi; McKenney, Anna Sophia; Meydan, Cem et al. (2017) Aid is a key regulator of myeloid/erythroid differentiation and DNA methylation in hematopoietic stem/progenitor cells. Blood 129:1779-1790
Nabel, Christopher S; DeNizio, Jamie E; Carroll, Martin et al. (2017) DNA Methyltransferases Demonstrate Reduced Activity against Arabinosylcytosine: Implications for Epigenetic Instability in Acute Myeloid Leukemia. Biochemistry 56:2166-2169
Glass, Jacob L; Hassane, Duane; Wouters, Bas J et al. (2017) Epigenetic Identity in AML Depends on Disruption of Nonpromoter Regulatory Elements and Is Affected by Antagonistic Effects of Mutations in Epigenetic Modifiers. Cancer Discov 7:868-883
Farge, Thomas; Saland, Estelle; de Toni, Fabienne et al. (2017) Chemotherapy-Resistant Human Acute Myeloid Leukemia Cells Are Not Enriched for Leukemic Stem Cells but Require Oxidative Metabolism. Cancer Discov 7:716-735
Li, Sheng; Garrett-Bakelman, Francine E; Chung, Stephen S et al. (2016) Distinct evolution and dynamics of epigenetic and genetic heterogeneity in acute myeloid leukemia. Nat Med 22:792-9
Garrett-Bakelman, Francine E; Melnick, Ari M (2016) Mutant IDH: a targetable driver of leukemic phenotypes linking metabolism, epigenetics and transcriptional regulation. Epigenomics 8:945-57
Li, Sheng; Garrett-Bakelman, Francine; Perl, Alexander E et al. (2014) Dynamic evolution of clonal epialleles revealed by methclone. Genome Biol 15:472