Abstract
Heparanase plays important roles in tumor angiogenesis. Our previous study demonstrated that hypoxic preconditioning (HPC) enhanced the angiogenic and therapeutic effects of mesenchymal stem cells (MSCs), effects that were paralleled by enhanced heparanase expression. This study was designed to elucidate the role of heparanase in the improved therapeutic properties of HPC-MSCs and to explore underlying mechanisms using an ischemic rat hind limb model. MSCs transfected with heparanase (MSC<sup>hpa</sup>) or empty vector (MSC<sup>null</sup>) were delivered by intramuscular injections to ischemic hind limbs. Hind limbs that received MSC<sup>hpa</sup> recovered blood flow more rapidly at 7 days and acquired higher capillary density at 14 days compared with MSC<sup>null</sup>. Conditioned medium from MSC<sup>hpa</sup> increased endothelial cell migration and promoted greater tube formation relative to that from the MSC<sup>null</sup> groups. Vascular endothelial growth factor receptor 2 (VEGFR2, Flk-1) and its downstream signaling pathway (p38MAPK/HSP27) were significantly increased in human umbilical vein endothelial cells (HUVECs) after treatment with MSC<sup>hpa</sup> conditioned medium. Each of these responses was decreased by cocultured with MSC<sup>hpa-KD</sup> conditioned medium. MSC<sup>hpa</sup> conditioned medium activated hypoxia-inducible factor-2α (HIF-2α) and increased in parallel the transcript level of Flk-1 as determined by chromatin immunoprecipitation-PCR and luciferase assays. Analyses of integrin expression revealed an important role for integrin β1 in the regulation of HIF-2α. All angiogenic effects of MSC<sup>hpa</sup> conditioned medium were abolished by knockdown of integrin β1, HIF-2α, and Flk-1 in HUVECs with selective shRNAs. These findings identify heparanse as a key regulator of angiogenesis by MSCs. We propose a novel pathway wherein heparanse sequentially activates integrin β1, HIF-2α, Flk-1, and p38MAPK/HSP27 with corresponding enhancement of angiogenesis.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1850-1862 |
| Number of pages | 13 |
| Journal | Stem Cells |
| Volume | 33 |
| Issue number | 6 |
| DOIs | |
| State | Published - Jun 1 2015 |
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Keywords
- Angiogenesis
- Heparanase
- Mesenchymal stem cell
ASJC Scopus subject areas
- Cell Biology
- Developmental Biology
- Molecular Medicine
Cite this
Heparanase released from mesenchymal stem cells activates integrin beta1/HIF-2alpha/Flk-1 signaling and promotes endothelial cell migration and angiogenesis. / Hu, Xinyang; Zhang, Ling; Jin, Jing; Zhu, Wei; Xu, Yinchuan; Wu, Yan; Wang, Yingchao; Chen, Han; Webster, Keith A; Chen, Huiqiang; Yu, Hong; Wang, Jian'an.
In: Stem Cells, Vol. 33, No. 6, 01.06.2015, p. 1850-1862.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Heparanase released from mesenchymal stem cells activates integrin beta1/HIF-2alpha/Flk-1 signaling and promotes endothelial cell migration and angiogenesis
AU - Hu, Xinyang
AU - Zhang, Ling
AU - Jin, Jing
AU - Zhu, Wei
AU - Xu, Yinchuan
AU - Wu, Yan
AU - Wang, Yingchao
AU - Chen, Han
AU - Webster, Keith A
AU - Chen, Huiqiang
AU - Yu, Hong
AU - Wang, Jian'an
PY - 2015/6/1
Y1 - 2015/6/1
N2 - Heparanase plays important roles in tumor angiogenesis. Our previous study demonstrated that hypoxic preconditioning (HPC) enhanced the angiogenic and therapeutic effects of mesenchymal stem cells (MSCs), effects that were paralleled by enhanced heparanase expression. This study was designed to elucidate the role of heparanase in the improved therapeutic properties of HPC-MSCs and to explore underlying mechanisms using an ischemic rat hind limb model. MSCs transfected with heparanase (MSChpa) or empty vector (MSCnull) were delivered by intramuscular injections to ischemic hind limbs. Hind limbs that received MSChpa recovered blood flow more rapidly at 7 days and acquired higher capillary density at 14 days compared with MSCnull. Conditioned medium from MSChpa increased endothelial cell migration and promoted greater tube formation relative to that from the MSCnull groups. Vascular endothelial growth factor receptor 2 (VEGFR2, Flk-1) and its downstream signaling pathway (p38MAPK/HSP27) were significantly increased in human umbilical vein endothelial cells (HUVECs) after treatment with MSChpa conditioned medium. Each of these responses was decreased by cocultured with MSChpa-KD conditioned medium. MSChpa conditioned medium activated hypoxia-inducible factor-2α (HIF-2α) and increased in parallel the transcript level of Flk-1 as determined by chromatin immunoprecipitation-PCR and luciferase assays. Analyses of integrin expression revealed an important role for integrin β1 in the regulation of HIF-2α. All angiogenic effects of MSChpa conditioned medium were abolished by knockdown of integrin β1, HIF-2α, and Flk-1 in HUVECs with selective shRNAs. These findings identify heparanse as a key regulator of angiogenesis by MSCs. We propose a novel pathway wherein heparanse sequentially activates integrin β1, HIF-2α, Flk-1, and p38MAPK/HSP27 with corresponding enhancement of angiogenesis.
AB - Heparanase plays important roles in tumor angiogenesis. Our previous study demonstrated that hypoxic preconditioning (HPC) enhanced the angiogenic and therapeutic effects of mesenchymal stem cells (MSCs), effects that were paralleled by enhanced heparanase expression. This study was designed to elucidate the role of heparanase in the improved therapeutic properties of HPC-MSCs and to explore underlying mechanisms using an ischemic rat hind limb model. MSCs transfected with heparanase (MSChpa) or empty vector (MSCnull) were delivered by intramuscular injections to ischemic hind limbs. Hind limbs that received MSChpa recovered blood flow more rapidly at 7 days and acquired higher capillary density at 14 days compared with MSCnull. Conditioned medium from MSChpa increased endothelial cell migration and promoted greater tube formation relative to that from the MSCnull groups. Vascular endothelial growth factor receptor 2 (VEGFR2, Flk-1) and its downstream signaling pathway (p38MAPK/HSP27) were significantly increased in human umbilical vein endothelial cells (HUVECs) after treatment with MSChpa conditioned medium. Each of these responses was decreased by cocultured with MSChpa-KD conditioned medium. MSChpa conditioned medium activated hypoxia-inducible factor-2α (HIF-2α) and increased in parallel the transcript level of Flk-1 as determined by chromatin immunoprecipitation-PCR and luciferase assays. Analyses of integrin expression revealed an important role for integrin β1 in the regulation of HIF-2α. All angiogenic effects of MSChpa conditioned medium were abolished by knockdown of integrin β1, HIF-2α, and Flk-1 in HUVECs with selective shRNAs. These findings identify heparanse as a key regulator of angiogenesis by MSCs. We propose a novel pathway wherein heparanse sequentially activates integrin β1, HIF-2α, Flk-1, and p38MAPK/HSP27 with corresponding enhancement of angiogenesis.
KW - Angiogenesis
KW - Heparanase
KW - Mesenchymal stem cell
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U2 - 10.1002/stem.1995
DO - 10.1002/stem.1995
M3 - Article
VL - 33
SP - 1850
EP - 1862
JO - Stem Cells
JF - Stem Cells
SN - 1066-5099
IS - 6
ER -