Characterization of the mechanisms involved in the early specification and migration of Prox1-expressing 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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P3.01-050 Isolation and Characterization of Lymphatic Endothelial Cells from Neoplastic and Normal Human Lung

Journal of Thoracic Oncology ◽

10.1016/j.jtho.2016.11.1616 ◽

2017 ◽

Vol 12 (1) ◽

pp. S1150

Author(s):

Federico Quaini ◽

Bruno Lorusso ◽

Angela Falco ◽

Denise Madeddu ◽

Costanza Lagrasta ◽

...

Keyword(s):

Endothelial Cells ◽

Human Lung ◽

Lymphatic Endothelial Cells ◽

Isolation And Characterization ◽

Normal Human

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Angiotensin II Stimulates the Proliferation and Migration of Lymphatic Endothelial Cells Through Angiotensin Type 1 Receptors

Frontiers in Physiology ◽

10.3389/fphys.2020.560170 ◽

2020 ◽

Vol 11 ◽

Author(s):

Qiu-Yue Lin ◽

Jie Bai ◽

Jin-Qiu Liu ◽

Hui-Hua Li

Keyword(s):

Endothelial Cells ◽

Angiotensin Ii ◽

Lymphatic Endothelial Cells ◽

And Migration ◽

Angiotensin Type ◽

Proliferation And Migration

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Isolation and characterization of primary lymphatic endothelial cells from rat thoracic duct

The FASEB Journal ◽

10.1096/fasebj.21.5.a619-c ◽

2007 ◽

Vol 21 (5) ◽

Author(s):

Hideki Ohta ◽

Kenichi Katsube ◽

Jyun‐ichi Ogawa ◽

Masaki Yanagishita

Keyword(s):

Endothelial Cells ◽

Thoracic Duct ◽

Lymphatic Endothelial Cells ◽

Isolation And Characterization

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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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P2‐3: Isolation and characterization of lymphatic endothelial cells from lung tissues affected by lymphangioleiomyomatosis

Respirology ◽

10.1111/resp.14150_99 ◽

2021 ◽

Vol 26 (S3) ◽

pp. 113-114

Keyword(s):

Endothelial Cells ◽

Lymphatic Endothelial Cells ◽

Isolation And Characterization

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FGFR Signaling Coordinates the Balance Between Glycolysis and Fatty Acid β-Oxidation in Lymphatic Endothelial Cells

10.1101/2020.05.20.107326 ◽

2020 ◽

Author(s):

Hongyuan Song ◽

Jie Zhu ◽

Longhou Fang ◽

Pengchun Yu

Keyword(s):

Endothelial Cells ◽

Fatty Acid ◽

Growth Factor Receptor ◽

Dependent Manner ◽

Lymphatic Endothelial Cells ◽

Energy Deficiency ◽

Atp Production ◽

Atp Generation ◽

And Migration ◽

A Minor

AbstractThe balance between glycolysis and oxidative phosphorylation is believed to be critical for maintaining cellular bioenergetics, yet the regulation of such balance in lymphatic endothelial cells (LECs) remains unclear. Here we found that chemical inhibition of fibroblast growth factor receptor (FGFR) activity or knockdown of FGFR1, which has been shown to suppress glycolysis and consequently ATP production, induces substantial upregulation of fatty acid β-oxidation (FAO), but not glucose oxidation or glutamine oxidation in LECs. Mechanistically, blockade of FGFR-AKT signaling promotes the expression of CPT1A, a rate-limiting enzyme of FAO, in a PPARα-dependent manner. Metabolic analysis further showed that CPT1A depletion impairs ATP generation in FGFR1-deficient rather than wild-type LECs. This result suggests that FAO, which makes a minor contribution to cellular energy under normal conditions, can compensate for energy deficiency caused by FGFR inhibition. Consequently, CPT1A silencing potentiates the effect of FGFR1 knockdown on impeding LEC proliferation and migration. Collectively, our study identified an FGFR-regulated metabolic balance that is important for LEC growth.

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