Project 4: MicroRNA targeting of normal and leukemia stem-progenitor cells (Civin) MicroRNAs bind to 3'untranslated region sites in target mRNAs to down-regulate translation to protein. Since microRNAs do not base-pair exactly with their target mRNAs, they can block protein translation of many mRNAs and serve as powerful switches to regulate cell functions. To extend our long-term studies on regulation of hematopoiesis and stem cell biology, we profiled microRNA expression in hematopoietic stemprogenitor cells (HSPCs). We combined this data with human HSPC mRNA expression results and microRNA-mRNA target predictions into a novel database which predicted that certain of the HSPCexpressed microRNAs (HE-microRNAs) targeted several mRNAs critical to hematopoiesis. On this informatic basis, we formulated a model for microRNA control of hematopoiesis in which many genes specifying hematopoietic differentiation are expressed by early HSPCs, but held in check by HE-microRNAs. For several target mRNAs, we then demonstrated experimentally that translation is actually decreased by microRNAs. Mir-155 potently reduced myeloid and erythroid colony formation of normal human HSPCs, and mir-16 selectively inhibited erythropoiesis. Since the cells that we studied include rare stem cells and various stages of progenitors, we propose in Aim 1, to expand our microRNA profiles of HSPCs to more highly purified subsets of primary human and mouse HSPCs and primary human acute myeloid leukemia (AMI) cells.
In Aim 2, we will determine if selected individual microRNAs experimentally inhibit development of primary HSPCs, as our model predicts, and if these microRNAs affect AMI stem cells and leukemogenesis. We will determine the proteins whose synthesis is inhibited by each functionally-active microRNA and thereby the molecular mechanisms of the hematopoietic effects. At this point in our studies, at least 2 of these microRNAs, mir-16 and mir-155 appear to be new regulators of normal hematopoietic and cancer stem cells. Relevance: We suggest that a major barrier impeding cure of the many cancer patients whom we cannot cure today is our failure to effectively attack and eliminate cancer stem cells. Specifically, we believe that development of more effective therapies in leukemia hinges on fuller understanding of the regulation of rare normal and leukemic hematopoietic stem cells. The studies in this project investigate the effects of new regulatory molecules called microRNAs, which appear to play previously unexpected, potent roles in control of blood formation and leukemia. Understanding the actions of these hematopoietic-regulatory microRNAs may provide new targets, for both expansion of normal stem cells and leukemia therapy.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA070970-14
Application #
8374743
Study Section
Special Emphasis Panel (ZCA1-RPRB-J)
Project Start
Project End
2014-01-31
Budget Start
2012-02-01
Budget End
2013-01-31
Support Year
14
Fiscal Year
2012
Total Cost
$312,332
Indirect Cost
$101,267
Name
Johns Hopkins University
Department
Type
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Gamper, Christopher J; Takemoto, Clifford M; Chen, Allen R et al. (2016) High-dose Cyclophosphamide is Effective Therapy for Pediatric Severe Aplastic Anemia. J Pediatr Hematol Oncol 38:627-635
Fox, Jennifer M; Moynihan, James R; Mott, Bryan T et al. (2016) Artemisinin-derived dimer ART-838 potently inhibited human acute leukemias, persisted in vivo, and synergized with antileukemic drugs. Oncotarget 7:7268-79
Kim, MinJung; Tan, Yee Sun; Cheng, Wen-Chih et al. (2015) MIR144 and MIR451 regulate human erythropoiesis via RAB14. Br J Haematol 168:583-97
Candia, Julián; Cherukuri, Srujana; Guo, Yin et al. (2015) Uncovering low-dimensional, miR-based signatures of acute myeloid and lymphoblastic leukemias with a machine-learning-driven network approach. Converg Sci Phys Oncol 1:
Rau, Rachel; Magoon, Daniel; Greenblatt, Sarah et al. (2014) NPMc+ cooperates with Flt3/ITD mutations to cause acute leukemia recapitulating human disease. Exp Hematol 42:101-13.e5
Brodsky, Robert A (2014) Paroxysmal nocturnal hemoglobinuria. Blood 124:2804-11
Tan, Yee Sun; Kim, MinJung; Kingsbury, Tami J et al. (2014) Regulation of RAB5C is important for the growth inhibitory effects of MiR-509 in human precursor-B acute lymphoblastic leukemia. PLoS One 9:e111777
Belet, Stefanie; Fieremans, Nathalie; Yuan, Xuan et al. (2014) Early frameshift mutation in PIGA identified in a large XLID family without neonatal lethality. Hum Mutat 35:350-5
Ma, Hayley S; Nguyen, Bao; Duffield, Amy S et al. (2014) FLT3 kinase inhibitor TTT-3002 overcomes both activating and drug resistance mutations in FLT3 in acute myeloid leukemia. Cancer Res 74:5206-17
Yuan, Xuan; Braunstein, Evan M; Ye, Zhaohui et al. (2013) Generation of glycosylphosphatidylinositol anchor protein-deficient blood cells from human induced pluripotent stem cells. Stem Cells Transl Med 2:819-29

Showing the most recent 10 out of 52 publications