Molecular and Cellular Effects of In Vitro Shockwave Treatment on Lymphatic Endothelial Cells

Abstract Lymphangioleiomyomatosis (LAM) is a rare pulmonary disease characterised by the proliferation of smooth muscle-like cells (LAM cells), and an abundance of lymphatic vessels in LAM lesions. Studies reported that vascular endothelial growth factor-D (VEGF-D) secreted by LAM cells contributes to LAM-associated lymphangiogenesis, however, the precise mechanisms of lymphangiogenesis and characteristics of lymphatic endothelial cells (LECs) in LAM lesions have not yet been elucidated. In this study, human primary-cultured LECs were obtained both from LAM-affected lung tissues (LAM-LECs) and normal lung tissues (control LECs) using fluorescence-activated cell sorting (FACS). We found that LAM-LECs had significantly higher ability of proliferation and migration compared to control LECs. VEGF-D significantly promoted migration of LECs but not proliferation of LECs in vitro. cDNA microarray and FACS analysis revealed the expression of vascular endothelial growth factor receptor (VEGFR)-3 and integrin α9 were elevated in LAM-LECs. Inhibition of VEGFR-3 suppressed proliferation and migration of LECs, and blockade of integrin α9 reduced VEGF-D-induced migration of LECs. Our data uncovered the distinct features of LAM-associated LECs, increased proliferation and migration, which may be due to higher expression of VEGFR-3 and integrin α9. Furthermore, we also found VEGF-D/VEGFR-3 and VEGF-D/ integrin α9 signaling play an important role in LAM-associated lymphangiogenesis.

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Role of synectin in lymphatic development in zebrafish and frogs

Blood ◽

10.1182/blood-2009-11-254557 ◽

2010 ◽

Vol 116 (17) ◽

pp. 3356-3366 ◽

Cited By ~ 30

Author(s):

Karlien Hermans ◽

Filip Claes ◽

Wouter Vandevelde ◽

Wei Zheng ◽

Ilse Geudens ◽

...

Keyword(s):

Endothelial Cells ◽

Thoracic Duct ◽

Pdz Domain ◽

Lymphatic Endothelial Cells ◽

Cardinal Vein ◽

Morpholino Knockdown ◽

Lymphatic Development ◽

Factor C ◽

Endothelial Growth Factor

Abstract The molecular basis of lymphangiogenesis remains incompletely characterized. Here, we document a novel role for the PDZ domain-containing scaffold protein synectin in lymphangiogenesis using genetic studies in zebrafish and tadpoles. In zebrafish, the thoracic duct arises from parachordal lymphangioblast cells, which in turn derive from secondary lymphangiogenic sprouts from the posterior cardinal vein. Morpholino knockdown of synectin in zebrafish impaired formation of the thoracic duct, due to selective defects in lymphangiogenic but not angiogenic sprouting. Synectin genetically interacted with Vegfr3 and neuropilin-2a in regulating lymphangiogenesis. Silencing of synectin in tadpoles caused lymphatic defects due to an underdevelopment and impaired migration of Prox-1+ lymphatic endothelial cells. Molecular analysis further revealed that synectin regulated Sox18-induced expression of Prox-1 and vascular endothelial growth factor C–induced migration of lymphatic endothelial cells in vitro. These findings reveal a novel role for synectin in lymphatic development.

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Adipose-Derived Stem Cells Promote Proliferation, Migration, and Tube Formation of Lymphatic Endothelial Cells In Vitro by Secreting Lymphangiogenic Factors

Annals of Plastic Surgery ◽

10.1097/sap.0000000000000084 ◽

2015 ◽

Vol 74 (6) ◽

pp. 728-736 ◽

Cited By ~ 35

Author(s):

Kohsuke Takeda ◽

Yoshihiro Sowa ◽

Kenichi Nishino ◽

Kyoko Itoh ◽

Shinji Fushiki

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Tube Formation ◽

Adipose Derived Stem Cells ◽

Lymphatic Endothelial Cells

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HMGB1 promotes lymphangiogenesis of human lymphatic endothelial cells in vitro

Medical Oncology ◽

10.1007/s12032-010-9778-7 ◽

2010 ◽

Vol 29 (1) ◽

pp. 358-363 ◽

Cited By ~ 7

Author(s):

Yuanyuan Qiu ◽

Ying Chen ◽

Xin Fu ◽

Lei Zhang ◽

Jing Tian ◽

...

Keyword(s):

Endothelial Cells ◽

Lymphatic Endothelial Cells

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Isolation and Characterization of Human Lung Lymphatic Endothelial Cells

BioMed Research International ◽

10.1155/2015/747864 ◽

2015 ◽

Vol 2015 ◽

pp. 1-12 ◽

Cited By ~ 4

Author(s):

Bruno Lorusso ◽

Angela Falco ◽

Denise Madeddu ◽

Caterina Frati ◽

Stefano Cavalli ◽

...

Keyword(s):

Endothelial Cells ◽

Tyrosine Kinases ◽

Human Lung ◽

Lymphatic Endothelial Cells ◽

Isolation And Characterization ◽

Microbial Agents ◽

Diseased Lung

Characterization of lymphatic endothelial cells from the respiratory system may be crucial to investigate the role of the lymphatic system in the normal and diseased lung. We describe a simple and inexpensive method to harvest, isolate, and expand lymphatic endothelial cells from the human lung (HL-LECs). Fifty-five samples of healthy lung selected from patients undergoing lobectomy were studied. A two-step purification tool, based on paramagnetic sorting with monoclonal antibodies to CD31 and Podoplanin, was employed to select a pure population of HL-LECs. The purity of HL-LECs was assessed by morphologic criteria, immunocytochemistry, flow cytometry, and functional assays. Interestingly, these cells retainin vitroseveral receptor tyrosine kinases (RTKs) implicated in cell survival and proliferation. HL-LECs represent a clinically relevant cellular substrate to study lymphatic biology, lymphoangiogenesis, interaction with microbial agents, wound healing, and anticancer therapy.

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Phomaketide A Inhibits Lymphangiogenesis in Human Lymphatic Endothelial Cells

Marine Drugs ◽

10.3390/md17040215 ◽

2019 ◽

Vol 17 (4) ◽

pp. 215 ◽

Cited By ~ 2

Author(s):

Huai-Ching Tai ◽

Tzong-Huei Lee ◽

Chih-Hsin Tang ◽

Lei-Po Chen ◽

Wei-Cheng Chen ◽

...

Keyword(s):

Endothelial Cells ◽

Protease Inhibitors ◽

Cancer Metastasis ◽

New Drugs ◽

Tumor Xenograft ◽

Specific Marker ◽

Lymphatic Endothelial Cells ◽

Matrix Metalloproteinase 1

Lymphangiogenesis is an important biological process associated with cancer metastasis. The development of new drugs that block lymphangiogenesis represents a promising therapeutic strategy. Marine fungus-derived compound phomaketide A, isolated from the fermented broth of Phoma sp. NTOU4195, has been reported to exhibit anti-angiogenic and anti-inflammatory effects. However, its anti-lymphangiogenic activity has not been clarified to date. In this study, we showed that phomaketide A inhibited cell growth, migration, and tube formation of lymphatic endothelial cells (LECs) without an evidence of cytotoxicity. Mechanistic investigations revealed that phomaketide A reduced LECs-induced lymphangiogenesis via vascular endothelial growth factor receptor-3 (VEGFR-3), protein kinase Cδ (PKCδ), and endothelial nitric oxide synthase (eNOS) signalings. Furthermore, human proteome array analysis indicated that phomaketide A significantly enhanced the protein levels of various protease inhibitors, including cystatin A, serpin B6, tissue factor pathway inhibitor (TFPI), and tissue inhibitor matrix metalloproteinase 1 (TIMP-1). Importantly, phomaketide A impeded tumor growth and lymphangiogenesis by decreasing the expression of LYVE-1, a specific marker for lymphatic vessels, in tumor xenograft animal model. These results suggest that phomaketide A may impair lymphangiogenesis by suppressing VEGFR-3, PKCδ, and eNOS signaling cascades, while simultaneously activating protease inhibitors in human LECs. We document for the first time that phomaketide A inhibits lymphangiogenesis both in vitro and in vivo, which suggests that this natural product could potentially treat cancer metastasis.

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Sphingosine 1-phosphate receptor 1 regulates the directional migration of lymphatic endothelial cells in response to fluid shear stress

Journal of The Royal Society Interface ◽

10.1098/rsif.2016.0823 ◽

2016 ◽

Vol 13 (125) ◽

pp. 20160823 ◽

Cited By ~ 6

Author(s):

Vinay N. Surya ◽

Eleftheria Michalaki ◽

Eva Y. Huang ◽

Gerald G. Fuller ◽

Alexander R. Dunn

Keyword(s):

Endothelial Cells ◽

Fluid Flow ◽

Molecular Mechanisms ◽

Fluid Shear Stress ◽

Lymphatic Vessels ◽

Upstream Migration ◽

Lymphatic Endothelial Cells ◽

Sphingosine 1 Phosphate ◽

G Protein Coupled

The endothelial cells that line blood and lymphatic vessels undergo complex, collective migration and rearrangement processes during embryonic development, and are known to be exquisitely responsive to fluid flow. At present, the molecular mechanisms by which endothelial cells sense fluid flow remain incompletely understood. Here, we report that both the G-protein-coupled receptor sphingosine 1-phosphate receptor 1 (S1PR1) and its ligand sphingosine 1-phosphate (S1P) are required for collective upstream migration of human lymphatic microvascular endothelial cells in an in vitro setting. These findings are consistent with a model in which signalling via S1P and S1PR1 are integral components in the response of lymphatic endothelial cells to the stimulus provided by fluid flow.

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Laminin γ2‐enriched extracellular vesicles of oral squamous cell carcinoma cells enhance in vitro lymphangiogenesis via integrin α3‐dependent uptake by lymphatic endothelial cells

International Journal of Cancer ◽

10.1002/ijc.32027 ◽

2019 ◽

Vol 144 (11) ◽

pp. 2795-2810 ◽

Cited By ~ 7

Author(s):

Ssu‐Han Wang ◽

Gunn‐Guang Liou ◽

Szu‐Heng Liu ◽

Jeffrey S. Chang ◽

Jenn‐Ren Hsiao ◽

...

Keyword(s):

Squamous Cell Carcinoma ◽

Endothelial Cells ◽

Oral Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Extracellular Vesicles ◽

Squamous Cell ◽

Carcinoma Cells ◽

Lymphatic Endothelial Cells

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ALK1 signaling regulates early postnatal lymphatic vessel development

Blood ◽

10.1182/blood-2009-07-235655 ◽

2010 ◽

Vol 115 (8) ◽

pp. 1654-1661 ◽

Cited By ~ 71

Author(s):

Kyle Niessen ◽

Gu Zhang ◽

John Brady Ridgway ◽

Hao Chen ◽

Minhong Yan

Keyword(s):

Endothelial Cells ◽

Transforming Growth Factor ◽

Vascular Development ◽

Lymphatic Vessels ◽

Transforming Growth Factor Β ◽

Type I ◽

Lymphatic Endothelial Cells ◽

Multiple Organs ◽

Lymphatic Development

Abstract In vertebrates, endothelial cells form 2 hierarchical tubular networks, the blood vessels and the lymphatic vessels. Despite the difference in their structure and function and genetic programs that dictate their morphogenesis, common signaling pathways have been recognized that regulate both vascular systems. ALK1 is a member of the transforming growth factor-β type I family of receptors, and compelling genetic evidence suggests its essential role in regulating blood vascular development. Here we report that ALK1 signaling is intimately involved in lymphatic development. Lymphatic endothelial cells express key components of the ALK1 pathway and respond robustly to ALK1 ligand stimulation in vitro. Blockade of ALK1 signaling results in defective lymphatic development in multiple organs of neonatal mice. We find that ALK1 signaling regulates the differentiation of lymphatic endothelial cells to influence the lymphatic vascular development and remodeling. Furthermore, simultaneous inhibition of ALK1 pathway increases apoptosis in lymphatic vessels caused by blockade of VEGFR3 signaling. Thus, our study reveals a novel aspect of ALK1 signaling in regulating lymphatic development and suggests that targeting ALK1 pathway might provide additional control of lymphangiogenesis in human diseases.

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