This proposal merges developmental and chemical biology, computational analysis, and bioengineering to tackle one of the most significant challenges in hematology research-the in vitro derivation of long-term engraftable hematopoietic stem cells from pluripotent stem cells. To date, considerable efforts at directed differentiation o hematopoietic stem cells (HSCs) from embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have proven only partially successful in mice and largely unsuccessful in humans. The failure to generate true ESC/iPSC-derived HSCs remains a barrier for exploiting patient-derived iPSCs for modeling blood diseases, and precludes harnessing this potentially transformative technology for therapy. This renewal application will build upon progress made during the first two years of the prior grant period to address the central question: What are the developmental pathways that drive the formation of HSCs in embryos, and how can we exploit this knowledge to direct the differentiation of pluripotent stem cells into clinically relevant hematopoietic lineages? We will apply computational algorithms to enhanced expression datasets generated by single cell RNA-seq of hematopoietic stem and progenitor populations from fish, mouse, and human. We will probe gene regulatory networks that direct hematopoietic and lymphoid development, and test candidate transcription factors, non-coding RNAs, and morphogens hypothesized to govern these networks, alone and in combination, through gain and loss-of-function strategies. In a second thrust, we will perform screens in zebrafish blastomeres and human pluripotent stem cells to discover novel chemical and biological regulators of HSCs, and will incorporate chemicals into cell differentiation protocols to drive HSC commitment and enhance HSC function. Lastly, using bioengineered platforms which mimic the biomechanical forces that act on the embryonic hemogenic endothelium, we will configure "hemogenic endothelium-on-a-chip" to interrogate the effectors and downstream signaling pathways that promote emergence of HSCs and expansion of hematopoietic stem and progenitor cells. Our efforts at defining genetic, chemical and biomechanical mechanisms in hematopoietic development represent complementary as well as synergistic strategies to solve our key challenge of deriving HSCs in vitro.

Public Health Relevance

This competitive renewal application merges developmental and chemical biology, computational analysis, and bioengineering to tackle one of the most significant challenges in hematology research-the in vitro derivation of long-term engraftable hematopoietic stem cells from pluripotent stem cells. We will address the central question: What are the developmental pathways that drive the formation of HSCs in embryos, and how can we exploit this knowledge to direct the differentiation of pluripotent stem cells into clinically releant hematopoietic lineages? Our efforts at defining genetic, chemical and biomechanical mechanisms in hematopoietic development represent complementary as well as synergistic strategies to solve our key challenge of deriving HSCs in vitro.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Resource-Related Research Projects (R24)
Project #
2R24DK092760-04
Application #
8771044
Study Section
Special Emphasis Panel (ZDK1-GRB-6 (M3))
Program Officer
Bishop, Terry Rogers
Project Start
2011-09-19
Project End
2019-07-31
Budget Start
2014-08-01
Budget End
2015-07-31
Support Year
4
Fiscal Year
2014
Total Cost
$1,449,467
Indirect Cost
$334,673
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
Arora, Natasha; Wenzel, Pamela L; McKinney-Freeman, Shannon L et al. (2014) Effect of developmental stage of HSC and recipient on transplant outcomes. Dev Cell 29:621-8
Morris, Samantha A; Cahan, Patrick; Li, Hu et al. (2014) Dissecting engineered cell types and enhancing cell fate conversion via CellNet. Cell 158:889-902
Hagedorn, Elliott J; Durand, Ellen M; Fast, Eva M et al. (2014) Getting more for your marrow: boosting hematopoietic stem cell numbers with PGE2. Exp Cell Res 329:220-6
Kumar, Roshan M; Cahan, Patrick; Shalek, Alex K et al. (2014) Deconstructing transcriptional heterogeneity in pluripotent stem cells. Nature 516:56-61
Cahan, Patrick; Li, Hu; Morris, Samantha A et al. (2014) CellNet: network biology applied to stem cell engineering. Cell 158:903-15
Doulatov, Sergei; Vo, Linda T; Chou, Stephanie S et al. (2013) Induction of multipotential hematopoietic progenitors from human pluripotent stem cells via respecification of lineage-restricted precursors. Cell Stem Cell 13:459-70
Tulpule, Asmin; Kelley, James M; Lensch, M William et al. (2013) Pluripotent stem cell models of Shwachman-Diamond syndrome reveal a common mechanism for pancreatic and hematopoietic dysfunction. Cell Stem Cell 12:727-36
Cherry, Anne B C; Daley, George Q (2013) Reprogrammed cells for disease modeling and regenerative medicine. Annu Rev Med 64:277-90
Cahan, Patrick; Daley, George Q (2013) Origins and implications of pluripotent stem cell variability and heterogeneity. Nat Rev Mol Cell Biol 14:357-68
Cesana, Marcella; Daley, George Q (2013) Deciphering the rules of ceRNA networks. Proc Natl Acad Sci U S A 110:7112-3

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