DOI
https://doi.org/10.25772/W57Q-9F90
Author ORCID Identifier
0000-0001-6357-8449
Defense Date
2020
Document Type
Dissertation
Degree Name
Doctor of Philosophy
Department
Human and Molecular Genetics
First Advisor
Gordon D. Ginder
Second Advisor
Joyce Lloyd
Third Advisor
Joseph Landry
Fourth Advisor
Senthil Radhakrishnan
Fifth Advisor
Tomasz Kordula
Abstract
Depletion of the ATPase component of the Nucleosome Remodeling and Deacetylase (NuRD) complex, CHD4, reduces acute myeloid leukemia (AML) cell survival. This study identified other NuRD components, as potential therapeutic targets for disrupting protein-protein interactions within NuRD. In addition to AML, we established that T-cell Acute Lymphoblastic Leukemia (T-ALL) cell lines responded similarly to CHD4 depletion.
Greater than 90% depletion of either MBD2 or MBD3 (the mutually exclusive two DNA binding NuRD paralogues) – was unremarkable, but complete depletion of MBD3 increased apoptosis and genotoxic sensitivity. Combined depletion of MBD-NuRD proteins augmented apoptosis observed with complete MBD3 depletion - indicating redundancy of MBD2 and MBD3. This effect was ‘rescued’ by add-back of wild-type, but not mutant MBD2.
The mutually exclusive GATAD2A and GATAD2B NuRD paralogues, that connect CHD4 to MBD2/3, through the CR1 domain of GATAD2 and the coiled CC domain of MBD2/3, were next interrogated. GATAD2A depletion was unremarkable, but GATAD2B depletion reduced cell survival (increased apoptosis, reduced cell proliferation and colony forming ability) in cell lines and primary AML samples, but not in CD34+ hematopoietic stem cells. Additionally, the relative transcript levels of GATAD2B and c-Myc were significantly correlated.
Next, we showed that a CR1 penetrating peptide dislodges GATAD2B and CHD4 from NuRD in cell lysates. Cell lines and primary samples treated with the peptide showed significantly increased apoptosis, but normal CD34+ progenitors. We conclude that targeting the coiled-coil interaction between MBD proteins and the GATAD2B-CHD4 chromatin remodeling subcomplex of NuRD may be a promising therapeutic strategy in acute leukemia.
Rights
© The Author
Is Part Of
VCU University Archives
Is Part Of
VCU Theses and Dissertations
Date of Submission
4-13-2020
Included in
Hematology Commons, Laboratory and Basic Science Research Commons, Molecular Genetics Commons, Neoplasms Commons