Abstract 4782: Epigenetic resistance to Notch inhibition in T cell acute lymphoblastic leukemia
Authors: Knoechel Birgit , Roderick Justine , Williamson Kaylyn , Zhu Jiang , Lohr Jens , Cotton Matthew , Gillespie Shawn , Fernandez Daniel , Ku Manching , Wang Hongfang , Piccioni Federica , Silver Serena , Jain Mohit , Pearson Daniel , Kluk Michael , Ott Christopher , Greiner Dale , Brehm Michael , Shultz Leonard , Gutierrez Alejandro , Stegmaier Kimberly , Harris Marian , Silverman Lewis , Sallan Stephen , Kung Andrew , Root David , Bradner James , Aster Jon , Kelliher Michelle , Bernstein Bradley 
Published: May 2015

Abstract
Resistance to therapy is one of the major challenges in cancer treatment today, equally applicable to conventional chemotherapy as well as targeted therapy. Malignant tumors have widespread epigenetic alterations including aberrant expression of chromatin modifiers in a wide variety of tumors and chromosomal translocations involving chromatin modifiers that can drive development of some cancers. In addition, cancer genome sequencing studies have identified frequent somatic alterations in many chromatin-regulating enzymes. Moreover, epigenetic changes have been implicated in the development of drug resistance.
T cell acute lymphoblastic leukemia (T-ALL) has a high rate of treatment-refractory disease and relapse that significantly lowers survival rates compared to other forms of ALL. The identification of activating somatic NOTCH1 mutations in over 50% of patients with T-ALL led to the development of γ-secretase inhibitors (GSI) that prevent cleavage and activation of NOTCH1. Although effective in vitro, the rapid development of resistance that develops with Notch inhibition in vivo has so far prevented the translation of these inhibitors into the clinical setting.
We have developed a model of therapeutic resistance to inhibition of Notch signaling in T-ALL. In this model, ‘persister’ cells readily expand in the presence of GSI and the absence of Notch signaling. Rare persister cells are pre-existing in naïve T-ALL populations. Intriguingly, in vitro resistance to NOTCH1 inhibitor therapy is reversible, suggesting that it is epigenetically mediated. When compared to GSI-sensitive cells, persisters are characterized by distinct signaling and gene expression programs, and demonstrate global chromatin compaction. Using a short-hairpin knock-down screen of ∼ 300 known chromatin regulators we identified the chromatin reader BRD4 as essential for persister T-ALL cells. BRD4 expression levels are upregulated in persister T-ALL cells. Genome-wide binding studies of BRD4 show that it is found at active regulatory elements in the genome that are associated with genes known to be important for cell proliferation, survival and signaling pathways in T-ALL, e. g. MYC and BCL2. Treatment of persisters with the BRD4 inhibitor JQ1 down-regulates expression of these target genes. Functionally, JQ1 treatment leads to growth arrest and apoptosis in persister T-ALL cells, at doses well tolerated by GSI-sensitive leukemia cells. Furthermore, combination therapy of GSI and JQ1 is significantly more effective over vehicle or single agent therapy for primary human T-ALLs in vitro and in vivo.
These studies demonstrate epigenetic heterogeneity as a basis of drug resistance in leukemia. We suggest that combination therapies that include targeting of chromatin regulators may hold great therapeutic promise for prevention and treatment of resistant disease.
Citation Format: Birgit Knoechel, Justine Roderick, Kaylyn Williamson, Jiang Zhu, Jens Lohr, Matthew Cotton, Shawn Gillespie, Daniel Fernandez, Manching Ku, Hongfang Wang, Federica Piccioni, Serena Silver, Mohit Jain, Daniel Pearson, Michael Kluk, Christopher Ott, Dale Greiner, Michael Brehm, Leonard Shultz, Alejandro Gutierrez, Kimberly Stegmaier, Marian Harris, Lewis Silverman, Stephen Sallan, Andrew Kung, David Root, James Bradner, Jon Aster, Michelle Kelliher, Bradley Bernstein. Epigenetic resistance to Notch inhibition in T cell acute lymphoblastic leukemia. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4782. doi:10.1158/1538-7445.AM2014-4782