The long-term goal of this proposal is to provide the training and support for the applicant to become an independent physician-scientist with a career focused on studying the genomics and epigenomics of acute leukemia. The goals for the applicant include: obtaining practical and didactic training in scientific methodology, refining technical skills, and publishing the outlined research in top-tier scientific journals. The long-tem goal of the research is to define the role of KDM6A in normal and leukemic hematopoiesis. KDM6A encodes a H3K27 histone demethylase on the X chromosome that is mutated in a wide range of human cancers, including acute myeloid leukemia. Inactivating mutations of Kdm6a are the most common acquired progression events in our mouse model of acute promyelocytic leukemia (APL). To define the functional consequences of Kdm6a inactivation, we propose the following aims:
Specific Aim 1 : We will analyze a Kdm6a conditional knockout mouse to define the role of Kdm6a in normal and leukemic hematopoiesis. We will cross Kdm6a conditional knockout mice (with LoxP sites flanking the 3rd exon) with Vav-Cre transgenic mice to selectively delete Kdm6a in the hematopoietic lineage. We will characterize normal hematopoiesis using competitive repopulation assays, flow-based analysis of hematopoietic progenitors, and serial replating experiments. For these experiments, we will analyze 6-8 week old male Vav-Cre mice hemizygous for the floxed Kdm6a allele, along with the appropriate control mice. We will establish a tumor watch to determine if Kdm6a inactivation is associated with spontaneous leukemia development or bone marrow failure. We will intercross Kdm6a conditional knockout mice (with the Vav-Cre allele) with Ctsg-PML-RARA mice to define changes in cell growth, differentiation, and self renewal, and we will determine whether Kdm6a deficiency alters APL latency, penetrance, or phenotype in a tumor watch study.
Specific Aim 2 : We will define the epigenetic consequences of Kdm6a inactivation using RNA-Seq and ChIP-Seq. We will perform RNA-Seq on mouse APL tumors with and without spontaneous Kdm6a deletions or mutations, and on RNA derived from bone marrow cells from young, male Vav-Cre mice hemizygous for the Kdm6a conditional allele, along with the appropriate control animals. We will perform ChIP-Seq on mouse APL tumors with and without spontaneous Kdm6a deletions or mutations. We will use an anti-H3(tri-methyl K27) antibody to determine if changes in H3K27 methylation occur, and also with an anti-H3(tri-methyl K4) antibody, since Kdm6a inactivation may alter H3K4 methylation. Finally, we will perform RNA-Seq and ChIP-Seq on selected cell populations and leukemias (if they develop) in the male Vav-Cre mice hemizygous for the Kdm6a conditional allele, and APL tumors that arise in Ctsg-PML-RARA mice with and without the engineered deletion of Kdm6a. We will therefore comprehensively identify and analyze the epigenetic and transcriptional alterations associated with Kdm6a inactivation in wild type vs. PML-RARA expressing hematopoietic and leukemia cells.

Public Health Relevance

The long-term goal of this proposal is to provide the mentored support necessary to train a physician-scientist to become an independent investigator with a career focused on studying histone methylation and epigenomics of acute leukemia. The proposed research will define the role of the H3K27 demethylase KDM6A in normal and leukemic hematopoiesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08CA166229-03
Application #
8856167
Study Section
Subcommittee I - Transistion to Independence (NCI)
Program Officer
Lim, Susan E
Project Start
2013-07-01
Project End
2016-06-30
Budget Start
2015-07-01
Budget End
2016-06-30
Support Year
3
Fiscal Year
2015
Total Cost
Indirect Cost
Name
Washington University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Duncavage, Eric J; Jacoby, Meagan A; Chang, Gue Su et al. (2018) Mutation Clearance after Transplantation for Myelodysplastic Syndrome. N Engl J Med 379:1028-1041
Wartman, Lukas D (2018) The future of cancer treatment using precision oncogenomics. Cold Spring Harb Mol Case Stud 4:
Campbell, Katie M; Lin, Tianxiang; Zolkind, Paul et al. (2018) Oral Cavity Squamous Cell Carcinoma Xenografts Retain Complex Genotypes and Intertumor Molecular Heterogeneity. Cell Rep 24:2167-2178
Brestoff, Jonathan R; Vessoni, Alexandre T; Brenner, Kirsten A et al. (2018) Acute graft-versus-host disease following lung transplantation in a patient with a novel TERT mutation. Thorax 73:489-492
Christopher, Matthew J; Petti, Allegra A; Rettig, Michael P et al. (2018) Immune Escape of Relapsed AML Cells after Allogeneic Transplantation. N Engl J Med 379:2330-2341
Miller, Christopher A; Tricarico, Christopher; Skidmore, Zachary L et al. (2018) A case of acute myeloid leukemia with promyelocytic features characterized by expression of a novel RARG-CPSF6 fusion. Blood Adv 2:1295-1299
Halstead, Angela M; Kapadia, Chiraag D; Bolzenius, Jennifer et al. (2017) Bladder-cancer-associated mutations in RXRA activate peroxisome proliferator-activated receptors to drive urothelial proliferation. Elife 6:
Spencer, David H; Russler-Germain, David A; Ketkar, Shamika et al. (2017) CpG Island Hypermethylation Mediated by DNMT3A Is a Consequence of AML Progression. Cell 168:801-816.e13
Duncavage, Eric J; Uy, Geoffrey L; Petti, Allegra A et al. (2017) Mutational landscape and response are conserved in peripheral blood of AML and MDS patients during decitabine therapy. Blood 129:1397-1401
Griffith, Malachi; Spies, Nicholas C; Krysiak, Kilannin et al. (2017) CIViC is a community knowledgebase for expert crowdsourcing the clinical interpretation of variants in cancer. Nat Genet 49:170-174

Showing the most recent 10 out of 16 publications