Human CD34+ hematopoietic precursor cells can readily be obtained from the peripheral blood of adults. Treatment of these cells with various cytokine cocktails can induce efficient production of cells differentiated along erythroid, megakaryocytic or myeloid lines. We find that different cytokine cocktails induce rapid and extensive patterns of change in gene expression that are different for different cytokine cocktails. The prominence of these changes suggests that they occur in a large fraction of the CD34+ cells and may modulate the patterns of differentiation of individual cells. The recent development of methods that permit global measurements of mRNA expression, and DNA methylation in single cells make it possible to investigate the role of cytokines in lineage promotion at a level of specificity hitherto unavailable. In the present application we propose to use a combination of single cell genomic scale analyses, and cell tagging and labeling approaches to establish if the same cell can respond differently to different cytokine treatments, and to investigate whether these different responses result in differentiation along alternate lineages. The ultimate goal is to determine the nature of early transcriptional responses to lineage specific cytokines and whether these are instructional as well as permissive for lineage specification.

Public Health Relevance

Multiple types of blood cells can develop from a common precursor cell. The quantities of each type of cell are influenced by extra-cellular substances known as cytokines. We propose to use new techniques for analysis of single cells to determine how heterogeneous these precursor cells are and to what extent the cytokines determine the path of differentiation of the precursor cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK100858-02
Application #
8735141
Study Section
Molecular and Cellular Hematology (MCH)
Program Officer
Bishop, Terry Rogers
Project Start
2013-09-15
Project End
2016-06-30
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Yale University
Department
Genetics
Type
Schools of Medicine
DUNS #
City
New Haven
State
CT
Country
United States
Zip Code
06510
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