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Inhibition of CD146 lessens uveal melanoma progression through reducing angiogenesis and vasculogenic mimicry

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Abstract

Purpose

Anti-angiogenesis drug therapy is ineffective in treating uveal melanoma since it only targets angiogenesis leaving vasculogenic mimicry aside. There is no effective clinical strategy targeting vasculogenic mimicry, yet. We show here that CD146 is a novel target to inhibit uveal melanoma progression since it regulates both uveal melanoma angiogenesis and vasculogenic mimicry activity.

Methods

CD146 inhibition was achieved with its specific siRNAs or antibody AA98. Tube formation and migration of primary human retinal microvascular endothelial cells and tube-like structure formation, migration, invasion of uveal melanoma cells were evaluated after CD146 inhibition. The underlying mechanisms were investigated by Western blot and immunofluorescence. Finally, uveal melanoma cells were injected subretinally into the eyes of nude mice and AA98 was administrated. Tumor size was revealed by H&E staining, and angiogenesis and vasculogenic mimicry were evaluated with CD31-PAS staining.

Results

CD146 inhibition induced declines in tube formation and migration of primary human retinal microvascular endothelial cells and tube-like structure formation of uveal melanoma cells. CD146 mediated VEGFR/AKT/p38/NF-κB and FAK/VE-cadherin signal cascades were partially responsible for these biological effects. CD146 blockade by siRNA or AA98 also resulted in inhibition of migration and invasion as well as EMT process of uveal melanoma cells. The physiological relevance of such declines was confirmed by showing that AA98 treatment markedly suppressed the tumor growth, angiogenesis and vasculogenic mimicry induced by implantation of uveal melanoma cells into the eyes of nude mice.

Conclusions

CD146 is a novel mediator of both angiogenesis and vasculogenic mimicry in uveal melanoma. Its antibody AA98 has the potency to be developed as a new antibody drug for treating uveal melanoma. Our results warrant further assessment of CD146 as a potential target to improve therapeutic management of uveal melanoma in a clinical setting.

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Data availability

All data generated or analyzed during this study are included in this article. The datasets and materials used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

AKT:

Protein kinase B

ANOVA:

Analysis of variance

CM:

Conditioned medium

DAB:

Diaminobenzidine

DMEM:

Dulbecco’s modified Eagle’s medium

ECM:

Endothelial cell medium

EMT:

Epithelial-mesenchymal transition

FAK:

Focal adhesion kinase

FBS:

Fetal bovine serum

H&E:

Hematoxylin and eosin

HRMEC:

Human retinal microvascular endothelial cell

HRP:

Horseradish peroxidase

MCAM:

Melanoma cell adhesion molecule

MVD:

Mean vessel density

NC:

Negative control

NF-κB:

Nuclear factor kappa B

PAS:

Periodic acid Schiff

VEGF:

Vascular endothelial growth factor

VEGFR:

Vascular endothelial growth factor receptor

VMD:

Mean value of vasculogenic mimicry density

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Acknowledgements

Not applicable.

Funding

This work was supported by Zhejiang Provincial Natural Science Foundation of China (LY21H120006, LY16H120009), the Project of State Key Laboratory of Ophthalmology, Optometry and Visual Science, Wenzhou Medical University (J02-20190201), and the Youth Innovation Promotion Association of Chinese Academy of Sciences (2018122).

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Authors and Affiliations

  1. School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, 270 Xueyuan Road, Wenzhou, 325037, Zhejiang, China

    Ronghan Zhang, Xiaogang Chen, Shengwen Chen, Jiajia Tang, Feng Chen, Yong Lin, Peter Sol Reinach, LiLi Tu, Jia Qu & Qiang Hou

  2. State Key Laboratory of Optometry, Ophthalmology and Visual Science, Wenzhou Medical University, Wenzhou, 325037, Zhejiang, China

    Ronghan Zhang, Xiaogang Chen, Shengwen Chen, Jiajia Tang, Feng Chen, Yong Lin, Peter Sol Reinach, LiLi Tu, Jia Qu & Qiang Hou

  3. Key Laboratory of Protein and Peptide Pharmaceutical, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China

    Xiyun Yan & Hongxia Duan

  4. College of Life Sciences, University of Chinese Academy of Sciences, Beijing, 100049, China

    Xiyun Yan

  5. Joint Laboratory of Nanozymes in Zhengzhou University, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, China

    Xiyun Yan

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  1. Ronghan Zhang
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Contributions

QH, JQ, and HD conceived and designed this project. RZ, XC, SC, JT, FC and YL performed experiments. RZ, PR, XY, LT, HD, JQ and QH analyzed the data and drafted the manuscript. All authors reviewed and edited the final manuscript version. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Hongxia Duan, Jia Qu or Qiang Hou.

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All studies and procedures involving human tissues were approved by the Wenzhou Medical University ethics committee and performed in compliance with the Declaration of Helsinki and national laws. Studies on animals were performed in compliance with the ARVO Statement for the use of animals in Ophthalmic and Visual Research and with the Institutional Animal Care and Use Committee policies and procedures of Wenzhou Medical University.

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Zhang, R., Chen, X., Chen, S. et al. Inhibition of CD146 lessens uveal melanoma progression through reducing angiogenesis and vasculogenic mimicry. Cell Oncol. 45, 557–572 (2022). https://doi.org/10.1007/s13402-022-00682-9

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