Utilizing Zebrafish to Identify Anti‐(Lymph)Angiogenic Compounds for Cancer Treatment: Promise and Future Challenges

Okuda, Kazuhide S.; Lee, Hui Mei; Velaithan, Vithya; Ng, Mei Fong; Patel, Vyomesh

doi:10.1111/micc.12289

Cited by 11 publications

(15 citation statements)

References 175 publications

(208 reference statements)

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“…The blood vessels that grow into the xenograft have been shown to form an abnormal network of varying vessel morphology typical of mammalian tumours (Zhao et al, 2011a). The xenograft can be imaged in vivo , allowing observation of the mechanisms driving tumour vascularisation, such as sprouting angiogenesis, vascular co-option and endothelial cell migration (Zhao et al, 2011a,b), and can be used to screen for novel anti-angiogenesis drugs (Okuda et al, 2016). Despite the successes of this model, the precise role of the xenografted tumour cells and tumour-associated immune cells, with respect to the stimulation of angiogenesis, has not yet been determined.…”

Section: Introductionmentioning

confidence: 99%

Macrophages enhance Vegfa-driven angiogenesis in an embryonic zebrafish tumour xenograft model

Britto

Wyroba

Chen

et al. 2018

Disease Models &Amp; Mechanisms

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Tumour angiogenesis has long been a focus of anti-cancer therapy; however, anti-angiogenic cancer treatment strategies have had limited clinical success. Tumour-associated myeloid cells are believed to play a role in the resistance of cancer towards anti-angiogenesis therapy, but the mechanisms by which they do this are unclear. An embryonic zebrafish xenograft model has been developed to investigate the mechanisms of tumour angiogenesis and as an assay to screen anti-angiogenic compounds. In this study, we used cell ablation techniques to remove either macrophages or neutrophils and assessed their contribution towards zebrafish xenograft angiogenesis by quantitating levels of graft vascularisation. The ablation of macrophages, but not neutrophils, caused a strong reduction in tumour xenograft vascularisation and time-lapse imaging demonstrated that tumour xenograft macrophages directly associated with the migrating tip of developing tumour blood vessels. Finally, we found that, although macrophages are required for vascularisation in xenografts that either secrete VEGFA or overexpress zebrafish vegfaa, they are not required for the vascularisation of grafts with low levels of VEGFA, suggesting that zebrafish macrophages can enhance Vegfa-driven tumour angiogenesis. The importance of macrophages to this angiogenic response suggests that this model could be used to further investigate the interplay between myeloid cells and tumour vascularisation.

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Section: Introductionmentioning

confidence: 99%

Macrophages enhance Vegfa-driven angiogenesis in an embryonic zebrafish tumour xenograft model

Britto

Wyroba

Chen

et al. 2018

Disease Models &Amp; Mechanisms

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“…These cells migrated ventrally to the dorsal aorta to form the thoracic duct, which previously had only been hypothesized and indirectly supported but not directly measured[58]. Other transgenic zebrafish lines with fluorescent vasculature are reviewed extensively by Okuda et al[57]. …”

Section: Lymphatic Imaging Using Transgenic Fluorescent Reporter Micementioning

confidence: 99%

“…Taking advantage of the optical clarity of zebrafish embryos, investigators have been able to observe a wide range of vascular phenomena, including sprouting, remodeling, endothelial proliferation, and lumen formation, which are more difficult to study in the opaque embryos of mice. In addition, many new anti-lymphangiogenic compounds have been discovered using this model[57]. Zebrafish are especially useful for live imaging of deep lymphatic structures, such as the thoracic duct.…”

Section: Lymphatic Imaging Using Transgenic Fluorescent Reporter Micementioning

confidence: 99%

“…Identification of zebrafish genes, i.e., kdrl in blood endothelial cells and lyve1b in veins and LECs, and the tagging of these genes with various fluorophores have made for a robust experimental model that can be easily used for in vivo live observations of vascular development. For instance, this model has been used to screen for anti-lymphangiogenic compounds against cancers[57]. …”

Section: Lymphatic Imaging Using Transgenic Fluorescent Reporter Micementioning

confidence: 99%

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Fluorescent reporter transgenic mice for in vivo live imaging of angiogenesis and lymphangiogenesis

et al. 2018

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The study of lymphangiogenesis is an emerging science that has revealed the lymphatic system as a central player in many pathological conditions including cancer metastasis, lymphedema, and organ graft rejection. A thorough understanding of the mechanisms of lymphatic growth will play a key role in the development of therapeutic strategies against these conditions. Despite the known potential of this field, the study of lymphatics has historically lagged behind that of hemangiogenesis. Until recently, significant strides in lymphatic studies were impeded by a lack of lymphatic-specific markers and suitable experimental models compared to those of the more immediately visible blood vasculature. Lymphangiogenesis has also been shown to be a key phenomenon in developmental biological processes, such as cell proliferation, guided migration, differentiation, and cell-to-cell communication, making lymphatic-specific visualization techniques highly desirable and desperately needed. Imaging modalities including immunohistochemistry and in situ hybridization are limited by the need to sacrifice animal models for tissue harvesting at every experimental time point. Moreover, the processes of mounting and staining harvested tissues may introduce artifacts that can confound results. These traditional methods for investigating lymphatic and blood vasculature are associated with several problems including animal variability (e.g., between mice) when replicating lymphatic growth environments and the cost concerns of prolonged, labor-intensive studies, all of which complicate the study of dynamic lymphatic processes. With the discovery of lymphatic-specific markers, researchers have been able to develop several lymphatic and blood vessel-specific, promoter-driven, fluorescent-reporter transgenic mice for visualization of lymphatics in vivo and in vitro. For instance, GFP, mOrange, tdTomato, and other fluorescent proteins can be expressed under control of a lymphatic-specific marker like Prospero-related homeobox 1 (Prox1), which is a highly conserved transcription factor for determining embryonic organogenesis in vertebrates that is implicated in lymphangiogenesis as well as several human cancers. Importantly, Prox1-null mouse embryos develop without lymphatic vessels. In human adults, Prox1 maintains lymphatic endothelial cells and upregulates proteins associated with lymphangiogenesis (e.g., VEGFR-3) and downregulates angiogenesis-associated gene expression (e.g., STAT6). To visualize lymphatic development in the context of angiogenesis, dual fluorescent-transgenic reporters, like Prox1-GFP/Flt1-DsRed mice, have been bred to characterize lymphatic and blood vessels simultaneously in vivo. In this review, we discuss the trends in lymphatic visualization and the potential usage of transgenic breeds in hemangiogenesis and lymphangiogenesis research to understand spatial and temporal correlations between vascular development and pathological progression.

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“…Other researchers have found that CP-7 presents the characteristics of low toxicity, high selectivity against VEGFR3, and potent inhibitory effect on VEGFR3-positive cancer cells both in vitro and in vivo. Practically, IMC-3C5 (NCT01288989), an anti-VEGFR3 antibody, has shown anti-BC activity in clinical practice. , To date, no VEGFR3-selective small-molecule inhibitor has been authorized by the FDA. It is necessary to identify a highly selective VEGFR3 inhibitor for the treatment of BC.…”

Section: Introductionmentioning

confidence: 99%

Discovery, Synthesis, and Evaluation of Highly Selective Vascular Endothelial Growth Factor Receptor 3 (VEGFR3) Inhibitor for the Potential Treatment of Metastatic Triple-Negative Breast Cancer

Yang

Zhang

et al. 2021

J. Med. Chem.

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We herein report the identification, structural optimization, and structure−activity relationship of thieno[2,3d]pyrimidine derivatives as a novel kind of selective vascular endothelial growth factor receptor 3 (VEGFR3) inhibitors. N-(4-Chloro-3-(trifluoromethyl)phenyl)-4-(6-(4-(4-methylpiperazin-1yl)phenyl)thieno[2,3-d]pyrimidin-4-yl)piperazine-1-carboxamide (38k) was the most potent VEGFR3 inhibitor (IC 50 = 110.4 nM) among developed compounds. Compared with VEGFR1 and VEGFR2, VEGFR3 was approximately 100 times more selective. Here, compound 38k significantly inhibited proliferation and migration of VEGF-C-induced human dermal lymphatic endothelial cells (HDLEC), MDA-MB-231, and MDA-MB-436 cells by inactivating the VEGFR3 signaling pathway. Additionally, 38k induced cell apoptosis and a prolonged G1/S-phase in MDA-MB-231 and MDA-MB-436 cells. It also presented acceptable pharmacokinetic characteristics in Sprague-Dawley (SD) rats with an oral bioavailability of 30.9%. In the xenograft model in vivo, 38k effectively inhibited breast cancer growth by suppressing the VEGFR3 signaling pathway. 38k pronouncedly resisted the formation of pulmonary metastatic nodules in mice. Collectively, 38k may be a promising therapeutic agent of metastatic breast cancer.

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Utilizing Zebrafish to Identify Anti‐(Lymph)Angiogenic Compounds for Cancer Treatment: Promise and Future Challenges

Cited by 11 publications

References 175 publications

Macrophages enhance Vegfa-driven angiogenesis in an embryonic zebrafish tumour xenograft model

Macrophages enhance Vegfa-driven angiogenesis in an embryonic zebrafish tumour xenograft model

Fluorescent reporter transgenic mice for in vivo live imaging of angiogenesis and lymphangiogenesis

Discovery, Synthesis, and Evaluation of Highly Selective Vascular Endothelial Growth Factor Receptor 3 (VEGFR3) Inhibitor for the Potential Treatment of Metastatic Triple-Negative Breast Cancer

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