Abstract
Angiopoietin-like proteins (angptls) are capable of ex vivo expansion of mouse and human hematopoietic stem and progenitor cells (HSPCs). Despite this intriguing ability, their mechanism is unknown. In this study, we show that angptl2 overexpression is sufficient to expand definitive HSPCs in zebrafish embryos. Angptl1/2 are required for definitive hematopoiesis and vascular specification of the hemogenic endothelium. The loss-of-function phenotype is reminiscent of the notch mutant mindbomb (mib), and a strong genetic interaction occurs between angptls and notch. Overexpressing angptl2 rescues mib while overexpressing notch rescues angptl1/2 morphants. Gene expression studies in ANGPTL2-stimulated CD34(+) cells showed a strong MYC activation signature and myc overexpression in angptl1/2 morphants or mib restored HSPCs formation. ANGPTL2 can increase NOTCH activation in cultured cells and ANGPTL receptor interacted with NOTCH to regulate NOTCH cleavage. Together our data provide insight to the angptl-mediated notch activation through receptor interaction and subsequent activation of myc targets.
Keywords:
LILRB2; angiopoietin-like proteins; developmental biology; hematopoietic stem and progenitor cells; myc; notch; stem cells; zebrafish.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Angiopoietin-Like Protein 1
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Angiopoietin-Like Protein 2
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Angiopoietin-like Proteins
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Angiopoietins / genetics*
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Angiopoietins / metabolism
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Animals
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Animals, Genetically Modified
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Blotting, Western
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Cells, Cultured
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Gene Expression Profiling
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Gene Expression Regulation, Developmental
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Gene Knockdown Techniques
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HEK293 Cells
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Hematopoiesis / genetics
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Hematopoietic Stem Cells / metabolism*
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Humans
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K562 Cells
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / metabolism
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Mice
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Microscopy, Confocal
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Protein Binding
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RNA Interference
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Receptors, Immunologic / genetics
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Receptors, Immunologic / metabolism
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Receptors, Notch / genetics*
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Receptors, Notch / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Signal Transduction / genetics*
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Time-Lapse Imaging
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Zebrafish / embryology
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Zebrafish / genetics
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Zebrafish / metabolism
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Zebrafish Proteins / genetics*
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Zebrafish Proteins / metabolism
Substances
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ANGPTL2 protein, human
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Angiopoietin-Like Protein 1
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Angiopoietin-Like Protein 2
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Angiopoietin-like Proteins
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Angiopoietins
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Angptl1 protein, zebrafish
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LILRB2 protein, human
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Membrane Glycoproteins
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Receptors, Immunologic
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Receptors, Notch
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Zebrafish Proteins
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angptl2b protein, zebrafish
Associated data
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GEO/GSE51652
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GEO/GSE63010