Efficient, inducible Cre‐recombinase activation in vascular endothelium

Claxton, Suzanne; Kostourou, Vassiliki; Jadeja, Shalini; Chambon, Pierre; Hodivala‐Dilke, Kairbaan; Fruttiger, Marcus

doi:10.1002/dvg.20367

Cited by 264 publications

(303 citation statements)

References 21 publications

(24 reference statements)

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“…To generate inducible, endothelialspecific mouse mutants for VEGFR2 and VEGFR3, we crossbred PdgfbiCreER T2 (21) and Cdh5-PAC-CreER T2 (22) mice with mice homozygous for the conditional Vegfr2 (23) (Vegfr2 flox/flox ) Significance VEGF/VEGFR and Notch signaling pathways are major molecular regulators of sprouting angiogenesis. Here we investigated the cross-talk between VEGFR and Notch signaling by using state-of-the-art inducible genetic mouse models.…”

Section: Resultsmentioning

confidence: 99%

VEGFR3 does not sustain retinal angiogenesis without VEGFR2

Zarkada

Heinolainen

Mäkinen

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

113

101

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Angiogenesis, the formation of new blood vessels, is regulated by vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs). VEGFR2 is abundant in the tip cells of angiogenic sprouts, where VEGF/VEGFR2 functions upstream of the delta-like ligand 4 (DLL4)/Notch signal transduction pathway. VEGFR3 is expressed in all endothelia and is indispensable for angiogenesis during early embryonic development. In adults, VEGFR3 is expressed in angiogenic blood vessels and some fenestrated endothelia. VEGFR3 is abundant in endothelial tip cells, where it activates Notch signaling, facilitating the conversion of tip cells to stalk cells during the stabilization of vascular branches. Subsequently, Notch activation suppresses VEGFR3 expression in a negative feedback loop. Here we used conditional deletions and a Notch pathway inhibitor to investigate the cross-talk between VEGFR2, VEGFR3, and Notch in vivo. We show that postnatal angiogenesis requires VEGFR2 signaling also in the absence of Notch or VEGFR3, and that even small amounts of VEGFR2 are able to sustain angiogenesis to some extent. We found that VEGFR2 is required independently of VEGFR3 for endothelial DLL4 up-regulation and angiogenic sprouting, and for VEGFR3 functions in angiogenesis. In contrast, VEGFR2 deletion had no effect, whereas VEGFR3 was essential for postnatal lymphangiogenesis, and even for lymphatic vessel maintenance in adult skin. Knowledge of these interactions and the signaling functions of VEGFRs in blood vessels and lymphatic vessels is essential for the therapeutic manipulation of the vascular system, especially when considering multitargeted antiangiogenic treatments.

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Section: Resultsmentioning

confidence: 99%

VEGFR3 does not sustain retinal angiogenesis without VEGFR2

Zarkada

Heinolainen

Mäkinen

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

113

101

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“…Since our study, Dr. Nusse's group identified ECs lining the central vein as the source of Wnt2 and Wnt9b in regulating β‐catenin activation and target gene expression in pericentral hepatocytes 8. We also employed platelet‐derived growth factor i‐Cre‐ERT2 to generate EC‐Wls‐KO; this led to a mosaic recombination insufficient to draw concrete conclusions, likely due to a different transgenic line 26. Eventually, we employed transgenic mice expressing Cre recombinase under Lyve1 promoter to delete Wls from ECs in the liver.…”

Section: Discussionmentioning

confidence: 99%

Endothelial Wnts regulate β‐catenin signaling in murine liver zonation and regeneration: A sequel to the Wnt–Wnt situation

Preziosi

Okabe

Poddar

et al. 2018

Hepatology Communications

103

145

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β‐Catenin in hepatocytes, under the control of Wnts, regulates pericentral gene expression. It also contributes to liver regeneration (LR) after partial hepatectomy (PH) by regulating cyclin‐D1 gene expression as shown in the β‐catenin and Wnt coreceptors low‐density lipoprotein receptor‐related protein 5/6 conditional knockouts (KO). However, conditional deletion of Wntless (Wls), required for Wnt secretion, in hepatocytes, cholangiocytes, or macrophages lacked any impact on zonation, while Wls deletion in macrophages only marginally affected LR. Here, we address the contribution of hepatic endothelial cells (ECs) in zonation and LR by characterizing EC‐Wls‐KO generated by interbreeding Wls‐floxed and lymphatic vessel endothelial hyaluronan receptor (Lyve1)‐cre mice. These mice were also used to study LR after PH. While Lyve1 expression in adult liver is limited to sinusoidal ECs only, Lyve1‐cre mice bred to ROSA26‐Stopflox/flox‐enhanced yellow fluorescent protein (EYFP) mice showed EYFP labeling in sinusoidal and central vein ECs. EC‐Wls‐KO mice showed decreased liver weights; lacked glutamine synthetase, cytochrome P450 2e1, and cytochrome P450 1a2; and were resistant to acetaminophen‐induced liver injury. After PH, EC‐Wls‐KO showed quantitative and qualitative differences in cyclin‐D1 expression at 24‐72 hours, which led to a lower hepatocyte proliferation at 40 hours but a rebound increase by 72 hours. ECs and macrophages isolated from regenerating livers at 12 hours showed significant up‐regulation of Wnt2 and Wnt9b messenger RNA; these are the same two Wnts involved in baseline β‐catenin activity in pericentral hepatocytes. Conclusion: At baseline, ECs secrete Wnt proteins essential for β‐catenin activation in pericentral hepatocytes. During LR, sinusoidal and central vein ECs and secondarily macrophages secrete Wnt2, while predominantly central vein ECs and secondarily macrophages are the likely source of Wnt9b. This process spatiotemporally regulates β‐catenin activation in hepatocytes to induce cell proliferation. (Hepatology Communications 2018;2:845‐860)

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“…More than 60% of these mice developed spontaneous (mostly multicentric) hemangiosarcoma by the average age of 25 wk. To further restrict p53 loss to the vascular endothelial lineages, other endothelialspecific Cre lines that have temporal control of Cre activity, such as VE-cadherin-CreER(T2) 30 or PDGFB-iCreER(T2), 31 could be used to avoid Cre activity in the hemogenic endothelium during development: this would thereby restrict p53 loss to the neonatal or adult endothelium and avoid p53 loss in the hematopoietic system. 32 We expect that such mice will only develop hemangiosarcomas.…”

Section: Discussionmentioning

confidence: 99%

Loss of autocrine endothelial-derived VEGF significantly reduces hemangiosarcoma development in conditional p53-deficient mice

et al. 2014

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Malignant transformation of the endothelium is rare, and hemangiosarcomas comprise only 1% of all sarcomas. For this reason and due to the lack of appropriate mouse models, the genetic mechanisms of malignant endothelial transformation are poorly understood. Here, we describe a hemangiosarcoma mouse model generated by deleting p53 specifically in the endothelial and hematopoietic lineages. This strategy led to a high incidence of hemangiosarcoma, with an average latency of 25 weeks. To study the in vivo roles of autocrine or endothelial cell autonomous VEGF signaling in the initiation and/or progression of hemangiosarcomas, we genetically deleted autocrine endothelial sources of VEGF in this mouse model. We found that loss of even a single conditional VEGF allele results in substantial rescue from endothelial cell transformation. These findings highlight the important role of threshold levels of autocrine VEGF signaling in endothelial malignancies and suggest a new approach for hemangiosarcoma treatment using targeted autocrine VEGF inhibition

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Efficient, inducible Cre‐recombinase activation in vascular endothelium

Cited by 264 publications

References 21 publications

VEGFR3 does not sustain retinal angiogenesis without VEGFR2

VEGFR3 does not sustain retinal angiogenesis without VEGFR2

Endothelial Wnts regulate β‐catenin signaling in murine liver zonation and regeneration: A sequel to the Wnt–Wnt situation

Loss of autocrine endothelial-derived VEGF significantly reduces hemangiosarcoma development in conditional p53-deficient mice

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