Abstract
In the developing embryo, instructive guidance from the ventral endoderm secures cardiac program induction within the anterolateral mesoderm. Endoderm-guided cardiogenesis, however, has yet to be resolved at the proteome level. Here, through cardiopoietic priming of the endoderm with the reprogramming cytokine tumor necrosis factor α (TNFα), candidate effectors of embryonic stem cell cardiac differentiation were delineated by comparative proteomics. Differential two-dimensional gel electrophoretic mapping revealed that more than 75% of protein species increased >1.5-fold in the TNFα-primed versus unprimed endodermal secretome. Protein spot identification by linear ion trap quadrupole (LTQ) tandem mass spectrometry (MS/MS) and validation by shotgun LTQ-Fourier transform MS/MS following multidimensional chromatography mapped 99 unique proteins from 153 spot assignments. A definitive set of 48 secretome proteins was deduced by iterative bioinformatic screening using algorithms for detection of canonical and noncanonical indices of secretion. Protein-protein interaction analysis, in conjunction with respective expression level changes, revealed a nonstochastic TNFα-centric secretome network with a scale-free hierarchical architecture. Cardiovascular development was the primary developmental function of the resolved TNFα-anchored network. Functional cooperativity of the derived cardioinductive network was validated through direct application of the TNFα-primed secretome on embryonic stem cells, potentiating cardiac commitment and sarcomerogenesis. Conversely, inhibition of primary network hubs negated the procardiogenic effects of TNFα priming. Thus, proteomic cartography establishes a systems biology framework for the endodermal secretome network guiding stem cell cardiopoiesis.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 387-400 |
| Number of pages | 14 |
| Journal | Stem Cells |
| Volume | 26 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 2008 |
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Keywords
- Cardiopoiesis
- Embryonic stem cell
- Endoderm
- Multidimensional liquid chromatography- tandem mass spectrometry
- Network biology
- Proteomics
- Secretome
- Systems biology
- Two-dimensional gel electrophoresis
ASJC Scopus subject areas
- Cell Biology
Cite this
Cardioinductive network guiding stem cell differentiation revealed by proteomic cartography of tumor necrosis factor α-primed endodermal secretome. / Arrell, D. Kent; Niederländer, Nicolas J.; Faustino, Randolph S.; Behfar, Atta; Terzic, Andre.
In: Stem Cells, Vol. 26, No. 2, 02.2008, p. 387-400.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Cardioinductive network guiding stem cell differentiation revealed by proteomic cartography of tumor necrosis factor α-primed endodermal secretome
AU - Arrell, D. Kent
AU - Niederländer, Nicolas J.
AU - Faustino, Randolph S.
AU - Behfar, Atta
AU - Terzic, Andre
PY - 2008/2
Y1 - 2008/2
N2 - In the developing embryo, instructive guidance from the ventral endoderm secures cardiac program induction within the anterolateral mesoderm. Endoderm-guided cardiogenesis, however, has yet to be resolved at the proteome level. Here, through cardiopoietic priming of the endoderm with the reprogramming cytokine tumor necrosis factor α (TNFα), candidate effectors of embryonic stem cell cardiac differentiation were delineated by comparative proteomics. Differential two-dimensional gel electrophoretic mapping revealed that more than 75% of protein species increased >1.5-fold in the TNFα-primed versus unprimed endodermal secretome. Protein spot identification by linear ion trap quadrupole (LTQ) tandem mass spectrometry (MS/MS) and validation by shotgun LTQ-Fourier transform MS/MS following multidimensional chromatography mapped 99 unique proteins from 153 spot assignments. A definitive set of 48 secretome proteins was deduced by iterative bioinformatic screening using algorithms for detection of canonical and noncanonical indices of secretion. Protein-protein interaction analysis, in conjunction with respective expression level changes, revealed a nonstochastic TNFα-centric secretome network with a scale-free hierarchical architecture. Cardiovascular development was the primary developmental function of the resolved TNFα-anchored network. Functional cooperativity of the derived cardioinductive network was validated through direct application of the TNFα-primed secretome on embryonic stem cells, potentiating cardiac commitment and sarcomerogenesis. Conversely, inhibition of primary network hubs negated the procardiogenic effects of TNFα priming. Thus, proteomic cartography establishes a systems biology framework for the endodermal secretome network guiding stem cell cardiopoiesis.
AB - In the developing embryo, instructive guidance from the ventral endoderm secures cardiac program induction within the anterolateral mesoderm. Endoderm-guided cardiogenesis, however, has yet to be resolved at the proteome level. Here, through cardiopoietic priming of the endoderm with the reprogramming cytokine tumor necrosis factor α (TNFα), candidate effectors of embryonic stem cell cardiac differentiation were delineated by comparative proteomics. Differential two-dimensional gel electrophoretic mapping revealed that more than 75% of protein species increased >1.5-fold in the TNFα-primed versus unprimed endodermal secretome. Protein spot identification by linear ion trap quadrupole (LTQ) tandem mass spectrometry (MS/MS) and validation by shotgun LTQ-Fourier transform MS/MS following multidimensional chromatography mapped 99 unique proteins from 153 spot assignments. A definitive set of 48 secretome proteins was deduced by iterative bioinformatic screening using algorithms for detection of canonical and noncanonical indices of secretion. Protein-protein interaction analysis, in conjunction with respective expression level changes, revealed a nonstochastic TNFα-centric secretome network with a scale-free hierarchical architecture. Cardiovascular development was the primary developmental function of the resolved TNFα-anchored network. Functional cooperativity of the derived cardioinductive network was validated through direct application of the TNFα-primed secretome on embryonic stem cells, potentiating cardiac commitment and sarcomerogenesis. Conversely, inhibition of primary network hubs negated the procardiogenic effects of TNFα priming. Thus, proteomic cartography establishes a systems biology framework for the endodermal secretome network guiding stem cell cardiopoiesis.
KW - Cardiopoiesis
KW - Embryonic stem cell
KW - Endoderm
KW - Multidimensional liquid chromatography- tandem mass spectrometry
KW - Network biology
KW - Proteomics
KW - Secretome
KW - Systems biology
KW - Two-dimensional gel electrophoresis
UR - http://www.scopus.com/inward/record.url?scp=40949164677&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=40949164677&partnerID=8YFLogxK
U2 - 10.1634/stemcells.2007-0599
DO - 10.1634/stemcells.2007-0599
M3 - Article
VL - 26
SP - 387
EP - 400
JO - Stem Cells
JF - Stem Cells
SN - 1066-5099
IS - 2
ER -