Heparanase released from mesenchymal stem cells activates integrin beta1/HIF-2alpha/Flk-1 signaling and promotes endothelial cell migration and angiogenesis

Xinyang Hu, Ling Zhang, Jing Jin, Wei Zhu, Yinchuan Xu, Yan Wu, Yingchao Wang, Han Chen, Keith A Webster, Huiqiang Chen, Hong Yu, Jian'an Wang

Research output: Contribution to journalArticle

13 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)1850-1862
Number of pages13
JournalStem Cells
Volume33
Issue number6
DOIs
StatePublished - Jun 1 2015

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CD29 Antigens
Conditioned Culture Medium
Mesenchymal Stromal Cells
Cell Movement
Endothelial Cells
Integrins
Extremities
Human Umbilical Vein Endothelial Cells
Vascular Endothelial Growth Factor Receptor-2
Chromatin Immunoprecipitation
Intramuscular Injections
Therapeutic Uses
Luciferases
heparanase
Polymerase Chain Reaction
endothelial PAS domain-containing protein 1
Therapeutics

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 journalArticle

Hu, X, Zhang, L, Jin, J, Zhu, W, Xu, Y, Wu, Y, Wang, Y, Chen, H, Webster, KA, Chen, H, Yu, H & Wang, J 2015, 'Heparanase released from mesenchymal stem cells activates integrin beta1/HIF-2alpha/Flk-1 signaling and promotes endothelial cell migration and angiogenesis', Stem Cells, vol. 33, no. 6, pp. 1850-1862. https://doi.org/10.1002/stem.1995
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. / Heparanase released from mesenchymal stem cells activates integrin beta1/HIF-2alpha/Flk-1 signaling and promotes endothelial cell migration and angiogenesis. In: Stem Cells. 2015 ; Vol. 33, No. 6. pp. 1850-1862.
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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 (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.",
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