A recessive screen for genes regulating hematopoietic stem cells

Identification of genes that regulate the development, self-renewal, and differentiation of stem cells is of vital importance for understanding normal organogenesis and cancer; such knowledge also underpins regenerative medicine. Here we demonstrate that chemical mutagenesis of mice combined with ad...

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Bibliographic Details
Published in:Blood 2010-12, Vol.116 (26), p.5849-5858
Main Authors: Papathanasiou, Peter, Tunningley, Robert, Pattabiraman, Diwakar R., Ye, Ping, Gonda, Thomas J., Whittle, Belinda, Hamilton, Adam E., Cridland, Simon O., Lourie, Rohan, Perkins, Andrew C.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Biomarkers - metabolism
Cell Differentiation
Cells, Cultured
Embryo, Mammalian - cytology
Embryo, Mammalian - physiology
Female
Flow Cytometry
Gene Expression Profiling
Genes, Recessive
Hematologic and hematopoietic diseases
Hematopoiesis - physiology
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - physiology
Male
Medical sciences
Mice
Mice, Inbred BALB C
Mice, Inbred C3H
Mice, Inbred C57BL
Mutation - genetics
Phenotype
Proto-Oncogene Proteins c-myb - genetics
Proto-Oncogene Proteins c-myb - metabolism
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Summary:Identification of genes that regulate the development, self-renewal, and differentiation of stem cells is of vital importance for understanding normal organogenesis and cancer; such knowledge also underpins regenerative medicine. Here we demonstrate that chemical mutagenesis of mice combined with advances in hematopoietic stem cell reagents and genome resources can efficiently recover recessive mutations and identify genes essential for generation and proliferation of definitive hematopoietic stem cells and/or their progeny. We used high-throughput fluorescence-activated cell sorter to analyze 9 subsets of blood stem cells, progenitor cells, circulating red cells, and platelets in more than 1300 mouse embryos at embryonic day (E) 14.5. From 45 pedigrees, we recovered 6 strains with defects in definitive hematopoiesis. We demonstrate rapid identification of a novel mutation in the c-Myb transcription factor that results in thrombocythemia and myelofibrosis as proof of principal of the utility of our fluorescence-activated cell sorter–based screen. Such phenotype-driven approaches will provide new knowledge of the genes, protein interactions, and regulatory networks that underpin stem cell biology.
ISSN:0006-4971
1528-0020
1528-0020
DOI:10.1182/blood-2010-04-269951