Hematopoietic stem cells (HSCs) are the functional units of bone marrow transplantation, which is used in the treatment of variety of blood cell diseases including leukemia and autoimmune disorders. Clinical use of HSCs is limited by the fact that they are rare cells, occurring at a frequency of only <1/20,000 bone marrow cells. Efforts to expand HSCs prior to transplant by ex vivo culturing have proven challenging and thus far such efforts have not translated to the clinic. Thus there remains a clinical need to find alternative strategies for either expanding the numbers of HSCs, or generating HSCs de novo. Numerous studies have shown that it is possible to experimentally reprogram the cellular identity of one cell type to another by enforced expression of transcription factors involved in the specification of the target cell type. The long- term objective of this project is to use this experimental paradigm and express transcription factors involved in specification of HSC fate and function in committed blood cells to reprogram them back to an induced stem cell fate. In order to achieve this we must first identify the factors involved in specifying the fate and function of HSCs. This will be achieved using a large-scale expression profiling strategy designed to allow us to identify transcription factors uniquely expressed in HSCs compared to the other cells of the blood system. Our preliminary studies have identified 28 such genes. Once identified we will then clone these factors into an inducible lentiviral delivery system to allow us to combinatorially express these genes in committed blood cells. We will then test whether or not committed blood cells receiving such factors are able of function like HSCs using a wide variety of stem cell assays. Once reprogramming has been achieved, it will then be important to determine the minimal combination of factors capable of mediating reprogramming. We will further examine the fidelity of reprogramming by functional, molecular and epigenetic analyses. Successful identification of factors capable of reprogramming committed blood cells to an induced HSC fate has the potential to advance our basic understanding of hematopoietic stem cell biology, and contribute to their clinical utility in a number of ways. For example, identification of factors capable of reprogramming committed blood cells to HSCs could provide important insights the molecular mechanisms involved in specifying the fundamental stem cell properties of self-renewal and multi-potency. In terms of clinical application, if successful, our study opens the possibility of eventually generating patient- specific induced HSCs for use in bone marrow transplantation, or for the study of blood disorders.

Public Health Relevance

Although cellular reprogramming strategies have been used to experimentally generate a number of cell types, the generation of hematopoietic stem cells (HSCs) by reprogramming has not yet been described. The goal of our proposal is to identify transcriptional factors with enriched expression in HSCs, and then express these factors in committed blood cells combinatorially to identify factors capable of imparting stem cell properties onto these otherwise committed cells. If successful, our proposal has the potential to provide important mechanistic insight into the central stem cell properties of self-renewal and multi-potency. It would also provide a novel experimental paradigm for generating and studying HSCs.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL107630-01A1
Application #
8196485
Study Section
Molecular and Cellular Hematology (MCH)
Program Officer
Thomas, John
Project Start
2011-07-01
Project End
2016-03-31
Budget Start
2011-07-01
Budget End
2012-03-31
Support Year
1
Fiscal Year
2011
Total Cost
$528,750
Indirect Cost
Name
Immune Disease Institute, Inc.
Department
Type
DUNS #
059709394
City
Boston
State
MA
Country
United States
Zip Code
02115
Garrison, Brian S; Rybak, Adrian P; Beerman, Isabel et al. (2017) ZFP521 regulates murine hematopoietic stem cell function and facilitates MLL-AF9 leukemogenesis in mouse and human cells. Blood 130:619-624
Lee, Jungmin; Dykstra, Brad; Spencer, Joel A et al. (2017) mRNA-mediated glycoengineering ameliorates deficient homing of human stem cell-derived hematopoietic progenitors. J Clin Invest 127:2433-2437
Gutierrez-Martinez, Paula; Rossi, Derrick J; Beerman, Isabel (2016) DNA Damage and Aging Around the Clock. Trends Mol Med 22:635-7
Dykstra, Brad; Lee, Jungmin; Mortensen, Luke J et al. (2016) Glycoengineering of E-Selectin Ligands by Intracellular versus Extracellular Fucosylation Differentially Affects Osteotropism of Human Mesenchymal Stem Cells. Stem Cells 34:2501-2511
Palchaudhuri, Rahul; Saez, Borja; Hoggatt, Jonathan et al. (2016) Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol 34:738-45
Lee, Jungmin; Dykstra, Brad; Sackstein, Robert et al. (2015) Progress and obstacles towards generating hematopoietic stem cells from pluripotent stem cells. Curr Opin Hematol 22:317-23
Beerman, Isabel; Rossi, Derrick J (2015) Epigenetic Control of Stem Cell Potential during Homeostasis, Aging, and Disease. Cell Stem Cell 16:613-25
Beerman, Isabel; Rossi, Derrick J (2014) Epigenetic regulation of hematopoietic stem cell aging. Exp Cell Res 329:192-9
Gazit, Roi; Mandal, Pankaj K; Ebina, Wataru et al. (2014) Fgd5 identifies hematopoietic stem cells in the murine bone marrow. J Exp Med 211:1315-31
Riddell, Jonah; Gazit, Roi; Garrison, Brian S et al. (2014) Reprogramming committed murine blood cells to induced hematopoietic stem cells with defined factors. Cell 157:549-64

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