The generation of desired cell types for therapeutic purposes is becoming a reality with the development of methods for deriving such cells from embryonic stem cells (ESC), induced pluripotent stem cells (iPSC), as well as from isolated adult stem/progenitor cells or differentiated cells that are directly """"""""reprogrammed"""""""" into lineage-specific stem/progenitor cells. Realizing this goal, however, will require methods for deriving therapeutically useful numbers of cells that avoid inducing permanent genetic alterations, and ensure the behavioral fidelity of derived lineage-committed stem/progenitor cells. To address this issue, we propose to test our hypotheses that pathways regulating normal development can be manipulated to direct differentiation and expansion of populations of cell types that reflect normal developmental states. As a model for this approach, we focus on the Notch and Wnt pathways and the well-characterized hematopoietic system to generate hematopoietic stem cells (HSC). The feasibility of expanding therapeutically useful stem/progenitor cells is demonstrated by our expansion of cord blood-derived stem/progenitor cells and by our successful application of these cells in a clinical setting. Specifically, we will examine the requirement for and timing of Notch and Wnt signaling in generating the first HSCs in the embryo, to guide our efforts to produce these cells ex vivo. We will generate ES- and iPS-derived HSC by enhancing differentiation towards hemogenic endothelial precursors of definitive hematopoietic stem/progenitor cells, and by promoting selfrenewal of these multipotent stem/progenitor populations (Project 1). To assess the therapeutic usefulness of human ES- and iPS-derived stem/progenitor cells, we will determine their preservation of the transcriptional, chromatin and DNA methylation and functional landscapes (Project 2). These studies will interface with those described in the collaborative linked application on the role of Wnt and Notch in expansion and proper differentiation of cardiac stem cells.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01HL100395-06
Application #
8656742
Study Section
Special Emphasis Panel (ZHL1)
Program Officer
Mondoro, Traci
Project Start
2009-09-30
Project End
2016-04-30
Budget Start
2014-05-01
Budget End
2015-04-30
Support Year
6
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
City
Seattle
State
WA
Country
United States
Zip Code
98109
Xu, Zhuojin; Robitaille, Aaron M; Berndt, Jason D et al. (2016) Wnt/β-catenin signaling promotes self-renewal and inhibits the primed state transition in naïve human embryonic stem cells. Proc Natl Acad Sci U S A 113:E6382-E6390
Kueh, Hao Yuan; Yui, Mary A; Ng, Kenneth K H et al. (2016) Asynchronous combinatorial action of four regulatory factors activates Bcl11b for T cell commitment. Nat Immunol 17:956-65
Li, Li B; Ma, Chao; Awong, Geneve et al. (2016) Silent IL2RG Gene Editing in Human Pluripotent Stem Cells. Mol Ther 24:582-91
van Galen, Peter; Viny, Aaron D; Ram, Oren et al. (2016) A Multiplexed System for Quantitative Comparisons of Chromatin Landscapes. Mol Cell 61:170-80
Rotem, Assaf; Ram, Oren; Shoresh, Noam et al. (2015) High-Throughput Single-Cell Labeling (Hi-SCL) for RNA-Seq Using Drop-Based Microfluidics. PLoS One 10:e0116328
Clarke, Raedun L; Robitaille, Aaron M; Moon, Randall T et al. (2015) A Quantitative Proteomic Analysis of Hemogenic Endothelium Reveals Differential Regulation of Hematopoiesis by SOX17. Stem Cell Reports 5:291-304
Rotem, Assaf; Ram, Oren; Shoresh, Noam et al. (2015) Single-cell ChIP-seq reveals cell subpopulations defined by chromatin state. Nat Biotechnol 33:1165-72
Lin, Michelle I; Price, Emily N; Boatman, Sonja et al. (2015) Angiopoietin-like proteins stimulate HSPC development through interaction with notch receptor signaling. Elife 4:
Ditadi, Andrea; Sturgeon, Christopher M; Tober, Joanna et al. (2015) Human definitive haemogenic endothelium and arterial vascular endothelium represent distinct lineages. Nat Cell Biol 17:580-91
Dahlberg, A; Woo, S; Delaney, C et al. (2015) Notch-mediated expansion of cord blood progenitors: maintenance of transcriptional and epigenetic fidelity. Leukemia 29:1948-51

Showing the most recent 10 out of 36 publications