Context- and Cell-Dependent Effects of Delta-Like 4 Targeting in the Bone Marrow Microenvironment

Leonor Remédio, Tânia Carvalho, Francisco Caiado, Ana Bastos-Carvalho, Diana Martins, António Duarte, Hideo Yagita, Sergio Dias

Research output: Contribution to journalArticle

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Abstract

Delta-like 4 (Dll4) is a ligand of the Notch pathway family which has been widely studied in the context of tumor angiogenesis, its blockade shown to result in non-productive angiogenesis and halted tumor growth. As Dll4 inhibitors enter the clinic, there is an emerging need to understand their side effects, namely the systemic consequences of Dll4:Notch blockade in tissues other than tumors. The present study focused on the effects of systemic anti-Dll4 targeting in the bone marrow (BM) microenvironment. Here we show that Dll4 blockade with monoclonal antibodies perturbs the BM vascular niche of sub-lethally irradiated mice, resulting in increased CD31+, VE-Cadherin+ and c-kit+ vessel density, and also increased megakaryocytes, whereas CD105+, VEGFR3+, SMA+ and lectin+ vessel density remained unaltered. We investigated also the expression of angiocrine genes upon Dll4 treatment in vivo, and demonstrate that IGFbp2, IGFbp3, Angpt2, Dll4, DHH and VEGF-A are upregulated, while FGF1 and CSF2 are reduced. In vitro treatment of endothelial cells with anti-Dll4 reduced Akt phosphorylation while maintaining similar levels of Erk 1/2 phosphorylation. Besides its effects in the BM vascular niche, anti-Dll4 treatment perturbed hematopoiesis, as evidenced by increased myeloid (CD11b+), decreased B (B220+) and T (CD3+) lymphoid BM content of treated mice, with a corresponding increase in myeloid circulating cells. Moreover, anti-Dll4 treatment also increased the number of CFU-M and -G colonies in methylcellulose assays, independently of Notch1. Finally, anti-Dll4 treatment of donor BM improved the hematopoietic recovery of lethally irradiated recipients in a transplant setting. Together, our data reveals the hematopoietic (BM) effects of systemic anti-Dll4 treatment result from qualitative vascular changes and also direct hematopoietic cell modulation, which may be favorable in a transplant setting.

Original languageEnglish
Article numbere52450
Pages (from-to)Online
JournalPlosOne
Volume7
Issue number12
DOIs
Publication statusPublished - 20 Dec 2012

Fingerprint

bone marrow
Bone
Bone Marrow
blood vessels
Blood Vessels
Tumors
Transplants
Phosphorylation
cells
angiogenesis
neoplasms
phosphorylation
niches
vascular endothelial growth factor A
megakaryocytes
Fibroblast Growth Factor 1
Neoplasms
methylcellulose
Methylcellulose
hematopoiesis

Keywords

  • HEMATOPOIETIC STEM-CELL
  • ; SINUSOIDAL ENDOTHELIAL-CELLS
  • INHIBITS TUMOR-GROWTH
  • NOTCH LIGAND
  • IN-VIVO
  • PROGENITOR CELLS
  • VASCULAR NICHE
  • SELF-RENEWAL
  • INDUCED MOBILIZATION
  • LINEAGE COMMITMENT

Cite this

Remédio, Leonor ; Carvalho, Tânia ; Caiado, Francisco ; Bastos-Carvalho, Ana ; Martins, Diana ; Duarte, António ; Yagita, Hideo ; Dias, Sergio. / Context- and Cell-Dependent Effects of Delta-Like 4 Targeting in the Bone Marrow Microenvironment. In: PlosOne. 2012 ; Vol. 7, No. 12. pp. Online.
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abstract = "Delta-like 4 (Dll4) is a ligand of the Notch pathway family which has been widely studied in the context of tumor angiogenesis, its blockade shown to result in non-productive angiogenesis and halted tumor growth. As Dll4 inhibitors enter the clinic, there is an emerging need to understand their side effects, namely the systemic consequences of Dll4:Notch blockade in tissues other than tumors. The present study focused on the effects of systemic anti-Dll4 targeting in the bone marrow (BM) microenvironment. Here we show that Dll4 blockade with monoclonal antibodies perturbs the BM vascular niche of sub-lethally irradiated mice, resulting in increased CD31+, VE-Cadherin+ and c-kit+ vessel density, and also increased megakaryocytes, whereas CD105+, VEGFR3+, SMA+ and lectin+ vessel density remained unaltered. We investigated also the expression of angiocrine genes upon Dll4 treatment in vivo, and demonstrate that IGFbp2, IGFbp3, Angpt2, Dll4, DHH and VEGF-A are upregulated, while FGF1 and CSF2 are reduced. In vitro treatment of endothelial cells with anti-Dll4 reduced Akt phosphorylation while maintaining similar levels of Erk 1/2 phosphorylation. Besides its effects in the BM vascular niche, anti-Dll4 treatment perturbed hematopoiesis, as evidenced by increased myeloid (CD11b+), decreased B (B220+) and T (CD3+) lymphoid BM content of treated mice, with a corresponding increase in myeloid circulating cells. Moreover, anti-Dll4 treatment also increased the number of CFU-M and -G colonies in methylcellulose assays, independently of Notch1. Finally, anti-Dll4 treatment of donor BM improved the hematopoietic recovery of lethally irradiated recipients in a transplant setting. Together, our data reveals the hematopoietic (BM) effects of systemic anti-Dll4 treatment result from qualitative vascular changes and also direct hematopoietic cell modulation, which may be favorable in a transplant setting.",
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Context- and Cell-Dependent Effects of Delta-Like 4 Targeting in the Bone Marrow Microenvironment. / Remédio, Leonor; Carvalho, Tânia ; Caiado, Francisco; Bastos-Carvalho, Ana; Martins, Diana; Duarte, António; Yagita, Hideo; Dias, Sergio.

In: PlosOne, Vol. 7, No. 12, e52450, 20.12.2012, p. Online.

Research output: Contribution to journalArticle

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AU - Remédio, Leonor

AU - Carvalho, Tânia

AU - Caiado, Francisco

AU - Bastos-Carvalho, Ana

AU - Martins, Diana

AU - Duarte, António

AU - Yagita, Hideo

AU - Dias, Sergio

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Y1 - 2012/12/20

N2 - Delta-like 4 (Dll4) is a ligand of the Notch pathway family which has been widely studied in the context of tumor angiogenesis, its blockade shown to result in non-productive angiogenesis and halted tumor growth. As Dll4 inhibitors enter the clinic, there is an emerging need to understand their side effects, namely the systemic consequences of Dll4:Notch blockade in tissues other than tumors. The present study focused on the effects of systemic anti-Dll4 targeting in the bone marrow (BM) microenvironment. Here we show that Dll4 blockade with monoclonal antibodies perturbs the BM vascular niche of sub-lethally irradiated mice, resulting in increased CD31+, VE-Cadherin+ and c-kit+ vessel density, and also increased megakaryocytes, whereas CD105+, VEGFR3+, SMA+ and lectin+ vessel density remained unaltered. We investigated also the expression of angiocrine genes upon Dll4 treatment in vivo, and demonstrate that IGFbp2, IGFbp3, Angpt2, Dll4, DHH and VEGF-A are upregulated, while FGF1 and CSF2 are reduced. In vitro treatment of endothelial cells with anti-Dll4 reduced Akt phosphorylation while maintaining similar levels of Erk 1/2 phosphorylation. Besides its effects in the BM vascular niche, anti-Dll4 treatment perturbed hematopoiesis, as evidenced by increased myeloid (CD11b+), decreased B (B220+) and T (CD3+) lymphoid BM content of treated mice, with a corresponding increase in myeloid circulating cells. Moreover, anti-Dll4 treatment also increased the number of CFU-M and -G colonies in methylcellulose assays, independently of Notch1. Finally, anti-Dll4 treatment of donor BM improved the hematopoietic recovery of lethally irradiated recipients in a transplant setting. Together, our data reveals the hematopoietic (BM) effects of systemic anti-Dll4 treatment result from qualitative vascular changes and also direct hematopoietic cell modulation, which may be favorable in a transplant setting.

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KW - PROGENITOR CELLS

KW - VASCULAR NICHE

KW - SELF-RENEWAL

KW - INDUCED MOBILIZATION

KW - LINEAGE COMMITMENT

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