Isolation and characterisation of lymphatic endothelial cells from lung tissues affected by lymphangioleiomyomatosis

• Published in: Scientific reports Vol. 11; no. 1; p. 8406
• Main Authors: Nishino, Koichi, Yoshimatsu, Yasuhiro, Muramatsu, Tomoki, Sekimoto, Yasuhito, Mitani, Keiko, Kobayashi, Etsuko, Okamoto, Shouichi, Ebana, Hiroki, Okada, Yoshinori, Kurihara, Masatoshi, Suzuki, Kenji, Inazawa, Johji, Takahashi, Kazuhisa, Watabe, Tetsuro, Seyama, Kuniaki
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16.04.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 631/114; 631/45; 631/80; 692/420; 692/699; Cell proliferation; DNA microarrays; Endothelial cells; Flow cytometry; Humanities and Social Sciences; Lesions; Lung diseases; Lymphatic system; multidisciplinary; Science; Science (multidisciplinary); Smooth muscle; Vascular endothelial growth factor; Vascular endothelial growth factor receptors
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-021-88064-3

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.