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Proteomic cornerstones of hematopoietic stem cell differentiation : distinct signatures of multipotent progenitors and myeloid committed cells

Klimmeck, Daniel; Hansson, Jenny LU ; Raffel, Simon; Vakhrushev, Sergey Y; Trumpp, Andreas and Krijgsveld, Jeroen (2012) In Molecular & Cellular Proteomics 11(8). p.286-302
Abstract

Regenerative tissues such as the skin epidermis, the intestinal mucosa or the hematopoietic system are organized in a hierarchical manner with stem cells building the top of this hierarchy. Somatic stem cells harbor the highest self-renewal activity and generate a series of multipotent progenitors which differentiate into lineage committed progenitors and subsequently mature cells. In this report, we applied an in-depth quantitative proteomic approach to analyze and compare the full proteomes of ex vivo isolated and FACS-sorted populations highly enriched for either multipotent hematopoietic stem/progenitor cells (HSPCs, Lin(neg)Sca-1(+)c-Kit(+)) or myeloid committed precursors (Lin(neg)Sca-1(-)c-Kit(+)). By employing stable isotope... (More)

Regenerative tissues such as the skin epidermis, the intestinal mucosa or the hematopoietic system are organized in a hierarchical manner with stem cells building the top of this hierarchy. Somatic stem cells harbor the highest self-renewal activity and generate a series of multipotent progenitors which differentiate into lineage committed progenitors and subsequently mature cells. In this report, we applied an in-depth quantitative proteomic approach to analyze and compare the full proteomes of ex vivo isolated and FACS-sorted populations highly enriched for either multipotent hematopoietic stem/progenitor cells (HSPCs, Lin(neg)Sca-1(+)c-Kit(+)) or myeloid committed precursors (Lin(neg)Sca-1(-)c-Kit(+)). By employing stable isotope dimethyl labeling and high-resolution mass spectrometry, more than 5000 proteins were quantified. From biological triplicate experiments subjected to rigorous statistical evaluation, 893 proteins were found differentially expressed between multipotent and myeloid committed cells. The differential protein content in these cell populations points to a distinct structural organization of the cytoskeleton including remodeling activity. In addition, we found a marked difference in the expression of metabolic enzymes, including a clear shift of specific protein isoforms of the glycolytic pathway. Proteins involved in translation showed a collective higher expression in myeloid progenitors, indicating an increased translational activity. Strikingly, the data uncover a unique signature related to immune defense mechanisms, centering on the RIG-I and type-1 interferon response systems, which are installed in multipotent progenitors but not evident in myeloid committed cells. This suggests that specific, and so far unrecognized, mechanisms protect these immature cells before they mature. In conclusion, this study indicates that the transition of hematopoietic stem/progenitors toward myeloid commitment is accompanied by a profound change in processing of cellular resources, adding novel insights into the molecular mechanisms at the interface between multipotency and lineage commitment.

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Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
keywords
Animals, Cell Differentiation, Female, Flow Cytometry, Glycolysis, Hematopoiesis, Hematopoietic Stem Cells, Immunohistochemistry, Mice, Mice, Inbred C57BL, Multipotent Stem Cells, Myeloid Cells, Proteome, Proteomics, Spectrometry, Mass, Electrospray Ionization, Tandem Mass Spectrometry
in
Molecular & Cellular Proteomics
volume
11
issue
8
pages
17 pages
publisher
American Society for Biochemistry and Molecular Biology
external identifiers
  • Scopus:84864805939
ISSN
1535-9484
DOI
10.1074/mcp.M111.016790
language
English
LU publication?
no
id
f9411057-33b7-4d8b-8046-bbc7f62256b5
date added to LUP
2016-08-25 09:51:59
date last changed
2016-11-06 04:39:41
@misc{f9411057-33b7-4d8b-8046-bbc7f62256b5,
  abstract     = {<p>Regenerative tissues such as the skin epidermis, the intestinal mucosa or the hematopoietic system are organized in a hierarchical manner with stem cells building the top of this hierarchy. Somatic stem cells harbor the highest self-renewal activity and generate a series of multipotent progenitors which differentiate into lineage committed progenitors and subsequently mature cells. In this report, we applied an in-depth quantitative proteomic approach to analyze and compare the full proteomes of ex vivo isolated and FACS-sorted populations highly enriched for either multipotent hematopoietic stem/progenitor cells (HSPCs, Lin(neg)Sca-1(+)c-Kit(+)) or myeloid committed precursors (Lin(neg)Sca-1(-)c-Kit(+)). By employing stable isotope dimethyl labeling and high-resolution mass spectrometry, more than 5000 proteins were quantified. From biological triplicate experiments subjected to rigorous statistical evaluation, 893 proteins were found differentially expressed between multipotent and myeloid committed cells. The differential protein content in these cell populations points to a distinct structural organization of the cytoskeleton including remodeling activity. In addition, we found a marked difference in the expression of metabolic enzymes, including a clear shift of specific protein isoforms of the glycolytic pathway. Proteins involved in translation showed a collective higher expression in myeloid progenitors, indicating an increased translational activity. Strikingly, the data uncover a unique signature related to immune defense mechanisms, centering on the RIG-I and type-1 interferon response systems, which are installed in multipotent progenitors but not evident in myeloid committed cells. This suggests that specific, and so far unrecognized, mechanisms protect these immature cells before they mature. In conclusion, this study indicates that the transition of hematopoietic stem/progenitors toward myeloid commitment is accompanied by a profound change in processing of cellular resources, adding novel insights into the molecular mechanisms at the interface between multipotency and lineage commitment.</p>},
  author       = {Klimmeck, Daniel and Hansson, Jenny and Raffel, Simon and Vakhrushev, Sergey Y and Trumpp, Andreas and Krijgsveld, Jeroen},
  issn         = {1535-9484},
  keyword      = {Animals,Cell Differentiation,Female,Flow Cytometry,Glycolysis,Hematopoiesis,Hematopoietic Stem Cells,Immunohistochemistry,Mice,Mice, Inbred C57BL,Multipotent Stem Cells,Myeloid Cells,Proteome,Proteomics,Spectrometry, Mass, Electrospray Ionization,Tandem Mass Spectrometry},
  language     = {eng},
  number       = {8},
  pages        = {286--302},
  publisher    = {ARRAY(0x86a7d28)},
  series       = {Molecular & Cellular Proteomics},
  title        = {Proteomic cornerstones of hematopoietic stem cell differentiation : distinct signatures of multipotent progenitors and myeloid committed cells},
  url          = {http://dx.doi.org/10.1074/mcp.M111.016790},
  volume       = {11},
  year         = {2012},
}