RAMP1 signaling in immune cells regulates inflammation-associated lymphangiogenesis


Calcitonin gene-related peptide (CGRP) regulates inflammation via signaling through receptor activity-modifying protein (RAMP) 1. Here, we investigated the role of RAMP1 signaling in growth of lymphatic vessels during inflammation. Lymphangiogenesis in the diaphragm of RAMP1-deficient (−/−) mice or their wild-type (WT) counterparts was induced by repeated intraperitoneal injection of lipopolysaccharide (LPS). Compared with WT mice, LPS-induced lymphangiogenesis in RAMP1−/− mice was suppressed. This was accompanied by the reduced expression of vascular endothelial growth factor (VEGF)-C and VEGF-D. The number of CD4+ cells in diaphragm tissue from WT mice was greater than RAMP1−/− mice. Removing CD4+ cells attenuated lymphangiogenesis and expression of VEGF-C and VEGF-D. CD4+ cells isolated from RAMP1−/− mice exhibited reduced expression of VEGF-C and VEGF-D. The number of CD11b+ cells from RAMP1−/− mice was higher than WT mice and was associated with the upregulated expression of genes related to pro-inflammatory macrophage phenotype and downregulation of reparative macrophage phenotype-related expression. When fluorescein isothiocyanate (FITC)-dextran was injected into the peritoneal cavity, the amount of residual FITC-dextran in WT mice was lower than that in RAMP1−/− mice. The present results suggest that RAMP1 signaling in immune cells plays a critical role in inflammation-related lymphangiogenesis; therefore, it represents a novel target for controlling lymphangiogenesis.

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Fig. 1: Deficient RAMP1 signaling suppresses lymphangiogenesis and pro-lymphangiogenic factors in diaphragm tissues during inflammation.
Fig. 2: Accumulation of CD4+ and CD11b+ cells in diaphragm tissue after LPS injection.
Fig. 3: Involvement of CD4+ T cells in lymphangiogenesis during inflammation.
Fig. 4: Effect of RAMP1 expression in macrophages on expression of genes associated with inflammation.
Fig. 5: Effects of RAMP1 expression in bone marrow macrophages on genes associated with inflammation and pro-lymphangiogenic factors.
Fig. 6: Loss of RAMP1 results in impaired drainage of peritoneal fluid.


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We thank Michiko Ogino and Kyoko Yoshikawa for technical assistance. This research was supported by research grants (18K16994 to TM, 18H02605 to MM, 19K07127 to KH, and 19K16297 to HM) from the Japanese Ministry of Education, Culture, Sports, Science, and Technology, and the Kitasato University Research Grant for Young Researchers.

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Correspondence to Masataka Majima.

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Tsuru, S., Ito, Y., Matsuda, H. et al. RAMP1 signaling in immune cells regulates inflammation-associated lymphangiogenesis. Lab Invest (2020). https://doi.org/10.1038/s41374-019-0364-0

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