Abstract

In the initial funding period, we made the following observations: 1), the leukemogenic activity of intracellular Notch11 (ICN1) correlates with its ability to activate the transcription factor CSL; to recruit co-activators such as mastermind-like1 (MAML1); drive T cell development; and to inhibit B cell development; 2) blockade of Notch signaling at the level of the plasma membrane or the ICN1/MAML/CSL nuclear complex suppresses the growth of Notch1 leukemia cell lines, suggesting a role for Notch1 in leukemia cell growth and survival; and 3), constitutively active Notch2 and Notch3 also cause T-ALL in our murine model, whereas Notch4 does not. We now propose to build on these findings to clarify ICN structure/leukemia relationships, identify target genes downstream of ICN that are important in Notch leukemia induction and maintenance, and establish the role of co-activator recruitment in leukemogenesis and lymphoid development. Based on our success in suppressing the growth of ICN1-induced T-ALLs with MAML1 peptides, we will conduct a high-throughput screen for small molecule inhibitors of the ICN1/CSL/MAML1 complex. Together, these studies will provide a deeper understanding of Notch-induced leukemia and move us closer to novel therapeutic interventions in this disease.
The specific aims of the proposal are: 1. To determine the structural requirements for transformation of T cell progenitors by ICNs 1.1 To Identify the ICN2 and ICN3 domains required for induction of T-ALL 1.2 To reconstitute the ICN4-transforming activity with domain swaps 2. To determine how ICN signals maintain Notch leukemia cell growth 2.1. To determine the domains of ICNs that are required for Notch leukemia cell growth 2.2. To determine the Notch-induced gene expression program that maintains Notch leukemia cell growth 2.3. To identify proteins/signals that are necessary/sufficient for Notch leukemia cell growth 3. To determine the importance of co-activator recruitment in T cell development and transformation 3.1. To identify domains of MAML1 that are essential for function in normal and transformed lymphoid cells 3.2. To identify and characterize small molecule inhibitors of CSL/ICN/MAML1 function

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA082308-06
Application #
6768686
Study Section
Special Emphasis Panel (ZRG1-CAMP (01))
Program Officer
Mufson, R Allan
Project Start
1999-09-17
Project End
2008-06-30
Budget Start
2004-07-01
Budget End
2005-06-30
Support Year
6
Fiscal Year
2004
Total Cost
$418,530
Indirect Cost

  Institution

Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115

  Publications

Rodig, Scott J; Payne, Ethan G; Degar, Barbara A et al. (2008) Aggressive Langerhans cell histiocytosis following T-ALL: clonally related neoplasms with persistent expression of constitutively active NOTCH1. Am J Hematol 83:116-21
Fung, Erik; Tang, Sai-Man Timothy; Canner, James P et al. (2007) Delta-like 4 induces notch signaling in macrophages: implications for inflammation. Circulation 115:2948-56
Chiang, Mark Y; Xu, Mina L; Histen, Gavin et al. (2006) Identification of a conserved negative regulatory sequence that influences the leukemogenic activity of NOTCH1. Mol Cell Biol 26:6261-71
Malecki, Michael J; Sanchez-Irizarry, Cheryll; Mitchell, Jennifer L et al. (2006) Leukemia-associated mutations within the NOTCH1 heterodimerization domain fall into at least two distinct mechanistic classes. Mol Cell Biol 26:4642-51
Weng, Andrew P; Ferrando, Adolfo A; Lee, Woojoong et al. (2004) Activating mutations of NOTCH1 in human T cell acute lymphoblastic leukemia. Science 306:269-71
Sanchez-Irizarry, Cheryll; Carpenter, Andrea C; Weng, Andrew P et al. (2004) Notch subunit heterodimerization and prevention of ligand-independent proteolytic activation depend, respectively, on a novel domain and the LNR repeats. Mol Cell Biol 24:9265-73
Nam, Yunsun; Weng, Andrew P; Aster, Jon C et al. (2003) Structural requirements for assembly of the CSL.intracellular Notch1.Mastermind-like 1 transcriptional activation complex. J Biol Chem 278:21232-9
Vardar, Didem; North, Christopher L; Sanchez-Irizarry, Cheryll et al. (2003) Nuclear magnetic resonance structure of a prototype Lin12-Notch repeat module from human Notch1. Biochemistry 42:7061-7
Robertson, N G; Hamaker, S A; Patriub, V et al. (2003) Subcellular localisation, secretion, and post-translational processing of normal cochlin, and of mutants causing the sensorineural deafness and vestibular disorder, DFNA9. J Med Genet 40:479-86
Kutok, Jeffery L; Aster, Jon C (2002) Molecular biology of anaplastic lymphoma kinase-positive anaplastic large-cell lymphoma. J Clin Oncol 20:3691-702

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