The symmetry of stem cell division is one of the most fundamental questions in stem cell biology, and a leading goal of our research is identification of the key metabolic pathways that regulate hematopoietic stem cell (HSC) fate. We hypothesize that lipid metabolism contributes to HSC maintenance through precise control of division patterns. Single-cell approaches have identified the enhanced clearance of damaged mitochondria by fatty acid oxidation as an important mechanism of the self-renewing expansion of HSCs. However, our understanding of the relationship between HSC self-renewal and lipid metabolism is limited, as analyses of individual HSC division patterns have been hindered by both the heterogeneity of available HSC-enriched fractions and the technical challenges of imaging HSC fate in vivo. In addition, the number of cells required for full metabolomics analysis of rare populations of HSCs has proven prohibitive. To examine the activity upstream of fatty acid oxidation in HSCs, we have generated hematopoietic-specific conditional knockout mice for key genes impacting fatty acid oxidation pathway and/or fatty acid flow. A new biosensor for assessment of fatty acid oxidation activity in live cells has likewise been established to determine the metabolic modes which are most relevant to the controlled equilibrium of HSCs, and the gene-expression oriented bioinformatics tool, graphite, has been adapted to identify specific metabolite-dependent pathways. In order to illuminate the behavior of individual HSCs in vivo, we have established new technical regimens which include prospective isolation of HSCs with high purity based on Tie2 positivity, a local transplantation technique which delivers a single HSC under multiphoton microscopy guidance into the bone marrow of a live mouse, and micropipette aspiration to extract single cells after division directly from the bone marrow for functional or transcriptomic assay. Our project will utilize these advances to test our hypothesis regarding the roles of lipid metabolism in HSC fate choice. This in turn will facilitate novel therapeutic strategies for shifting the division balance of HSCs toward self-renewal through metabolic manipulation, and possibly contribute to improved clinical outcomes after HSC transplantation for non-malignant blood diseases. Thus, the goals of this proposal are three-fold: (1) In Aim 1, we will investigate the function of mitochondrial fatty acid oxidation in HSC division symmetry and explore a potential source of fatty acids to fulfill the requirements of HSCs; (2) In Aim 2, we will use the biosensor to identify key downstream metabolic targets of fatty acid metabolism for HSC fate and explore the measurement of the cellular metabolome in HSCs; and (3) finally, we propose in Aim 3 to directly examine in vivo HSC division symmetry, and the resulting division balance of fatty acid oxidation-defective HSCs will show definitively the in vivo relevance of fatty acid metabolism to HSC fate. If successful, the proposed research will positively impact the HSC field by providing a deeper understanding of the metabolic cues governing HSC fate decisions.

Public Health Relevance

Hematopoietic stem cell (HSC) division results in either self-renewal or differentiation, with the balance between the two options directly impacting hematopoietic homeostasis, but precisely how the specific modes of metabolism controls HSC fates remains unclear. This project utilizes various genetic models and metabolic assessments as well as single-cell approaches to understand the metabolic cues governing HSC fate decisions, with a focus on lipid metabolism. The resulting insights into the metabolic requirements of cell division will have a major impact on HSC research as well as current clinical practice for hematological diseases, as they will identify new strategies for shifting the division balance of HSCs toward self-renewal through metabolic manipulation.

National Institute of Health (NIH)
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Research Project (R01)
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Special Emphasis Panel (ZRG1)
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Bishop, Terry Rogers
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Albert Einstein College of Medicine
United States
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Ito, Kyoko; Ito, Keisuke (2018) Hematopoietic stem cell fate through metabolic control. Exp Hematol 64:1-11
Bonora, Massimo; Ito, Kyoko; Morganti, Claudia et al. (2018) Membrane-potential compensation reveals mitochondrial volume expansion during HSC commitment. Exp Hematol 68:30-37.e1
Guarnerio, Jlenia; Mendez, Lourdes Maria; Asada, Noboru et al. (2018) A non-cell-autonomous role for Pml in the maintenance of leukemia from the niche. Nat Commun 9:66
Weiss, Cary N; Ito, Keisuke (2017) A Macro View of MicroRNAs: The Discovery of MicroRNAs and Their Role in Hematopoiesis and Hematologic Disease. Int Rev Cell Mol Biol 334:99-175
Turcotte, Raphaël; Alt, Clemens; Runnels, Judith M et al. (2017) Image-guided transplantation of single cells in the bone marrow of live animals. Sci Rep 7:3875
Ito, Kyoko; Ito, Keisuke (2016) Metabolism and the Control of Cell Fate Decisions and Stem Cell Renewal. Annu Rev Cell Dev Biol 32:399-409
Ito, Kyoko; Turcotte, Raphaël; Cui, Jinhua et al. (2016) Self-renewal of a purified Tie2+ hematopoietic stem cell population relies on mitochondrial clearance. Science 354:1156-1160
Sato, Hanae; Wheat, Justin C; Steidl, Ulrich et al. (2016) DNMT3A and TET2 in the Pre-Leukemic Phase of Hematopoietic Disorders. Front Oncol 6:187
Ito, Keisuke; Frenette, Paul S (2016) HSC Contribution in Making Steady-State Blood. Immunity 45:464-466
Weiss, Cary N; Ito, Keisuke (2015) DNA damage: a sensible mediator of the differentiation decision in hematopoietic stem cells and in leukemia. Int J Mol Sci 16:6183-201

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