Despite frequent initial response to chemotherapy, overall cure rates have remained below 20% in patients with AML for the past 45 years, and relapse continues to be the most common cause of death. Recent evidence has shown that the accumulation of stepwise genetic and epigenetic changes in HSC lead to the formation of pre- leukemic stem cells (pre-LSC) that play a pivotal role not only in disease origination but also in leukemia relapse. While the existence and essentiality of such pre-cancerous cell states has been demonstrated in mice and humans, very little is known about the molecular mechanisms driving pre-LSC formation and progression. The transcription factor PU.1 is frequently heterozygously mutated or otherwise impaired in patients with AML. We have recently reported a mouse model of preleukemic-to-AML progression molecularly driven by heterozygous PU.1 enhancer deletion. This novel model is characterized by definable, functionally altered pre- leukemic stem cells and closely resembles human disease in key molecular, cell biological, and phenotypic features, including disease heterogeneity. This model now permits the identification and functional study of mechanisms driving the formation and progression of pre-leukemic stem cells. Furthermore, we have obtained proof-of-concept that PU.1-low AML cells show an increased vulnerability to further PU.1 inhibition (as complete loss of PU.1 leads to stem cell and hematopoietic failure), and we have developed first-in-class small- molecule pharmacologic inhibitors of PU.1, which directly interfere with PU.1-chromatin binding. Strikingly, we found that PU.1 inhibition by shRNA or small molecules has significant inhibitory effects on AML cells, including at the level of leukemia-initiating cells, while only minimally affecting normal HSC. PU.1 inhibition thus represents a new potential strategy to target AML. Based on our recent findings we propose to: 1. Identify and study pathways that are functionally critical for pre- LSC formation and maintenance; 2. Identify pathways that trigger the preleukemic-to-leukemic ?switch?, and progression of pre-LSC to different AML subtypes (mature/immature/bi-lineage), thus causing disease heterogeneity; 3. Study the mechanisms underlying the anti-leukemic effects of PU.1 inhibition in AML cells. We will employ our novel murine AML preLSC-to-LSC transition model as well as primary human MDS/AML samples. We will longitudinally study molecular changes at the stem cell level and functionally test dysregulated candidate pathways in vitro and in vivo. Furthermore, we will use shRNA and our novel drugs to identify PU.1 targets that mediate the anti-leukemic effects of PU.1 inhibition in AML cells. In summary, our study will improve our molecular understanding of pre-cancerous/leukemic cell states and their progression to fully transformed AML. Furthermore, our study will extend our proof-of-concept and understanding of PU.1 inhibition as a novel therapeutic strategy for the treatment of AML and at the pre-LSC level, a completely new approach in AML with considerable translational potential.

Public Health Relevance

Relapse continues to be the most common cause of death in acute myeloid leukemia (AML) and cure rates have not improved in 45 years and remain below 20%. Recent evidence has shown the existence and critical role of so-called pre-cancerous/pre-leukemic stem cells (pre-LSC) in disease initiation and relapse, however, very little is known about the molecular mechanisms driving pre-LSC formation and progression and how they could potentially be targeted therapeutically. To address this critical knowledge gap, the proposed research will molecularly study pre-LSC through the use of a newly developed mouse preLSC-to-AML progression model as well as primary human patients' samples.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Duglas Tabor, Yvonne
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Albert Einstein College of Medicine
United States
Zip Code
Escudero, Silvia; Zaganjor, Elma; Lee, Susan et al. (2018) Dynamic Regulation of Long-Chain Fatty Acid Oxidation by a Noncanonical Interaction between the MCL-1 BH3 Helix and VLCAD. Mol Cell 69:729-743.e7
Carvajal, Luis A; Neriah, Daniela Ben; Senecal, Adrien et al. (2018) Dual inhibition of MDMX and MDM2 as a therapeutic strategy in leukemia. Sci Transl Med 10:
Antony-Debré, Iléana; Paul, Ananya; Leite, Joana et al. (2017) Pharmacological inhibition of the transcription factor PU.1 in leukemia. J Clin Invest 127:4297-4313
Reyna, Denis E; Garner, Thomas P; Lopez, Andrea et al. (2017) Direct Activation of BAX by BTSA1 Overcomes Apoptosis Resistance in Acute Myeloid Leukemia. Cancer Cell 32:490-505.e10