Mast cell hyperactivity underpins the development of oxygen-induced retinopathy
Kenshiro Matsuda, … , Akane Tanaka, Hiroshi Matsuda
Kenshiro Matsuda, … , Akane Tanaka, Hiroshi Matsuda
Published October 9, 2017
Citation Information: J Clin Invest. 2017;127(11):3987-4000. https://doi.org/10.1172/JCI89893.
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Research Article Angiogenesis Inflammation

Mast cell hyperactivity underpins the development of oxygen-induced retinopathy

Abstract

Mast cells are classically thought to play an important role in protection against helminth infections and in the induction of allergic diseases; however, recent studies indicate that these cells also contribute to neovascularization, which is critical for tissue remodeling, chronic inflammation, and carcinogenesis. Here, we demonstrate that mast cells are essential for sprouting angiogenesis in a murine model of oxygen-induced retinopathy (OIR). Although mouse strains lacking mast cells did not exhibit retinal neovascularization following hypoxia, these mice developed OIR following infusion of mast cells or after injection of mast cell tryptase (MCT). Relative hypoxia stimulated mast cell degranulation via transient receptor potential ankyrin 1. Subsequent surges in MCT stimulated retinal endothelial cells to produce monocyte chemotactic protein-1 (MCP1) and angiogenic factors, leading to sprouting angiogenesis. Mast cell stabilizers as well as specific tryptase and MCP1 inhibitors prevented the development of OIR in WT mice. Preterm infants with early retinopathy of prematurity had markedly higher plasma MCT levels than age-matched infants without disease, suggesting mast cells contribute to human disease. Together, these results suggest therapies that suppress mast cell activity should be further explored as a potential option for preventing eye diseases and subsequent blindness induced by neovascularization.

Authors

Kenshiro Matsuda, Noriko Okamoto, Masatoshi Kondo, Peter D. Arkwright, Kaoru Karasawa, Saori Ishizaka, Shinichi Yokota, Akira Matsuda, Kyungsook Jung, Kumiko Oida, Yosuke Amagai, Hyosun Jang, Eiichiro Noda, Ryota Kakinuma, Koujirou Yasui, Uiko Kaku, Yasuo Mori, Nobuyuki Onai, Toshiaki Ohteki, Akane Tanaka, Hiroshi Matsuda

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Figure 4

Oxygen-induced degranulation of mast cells was induced by TRPA1.

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Oxygen-induced degranulation of mast cells was induced by TRPA1. (A) Whe...
(A) When BMCMCs were moved from 75% oxygen to 20% oxygen, significant degranulation was induced. TRPA1 inhibitor HC-030031 or TRPA1 deficiency suppressed relative hypoxia-induced degranulation of BMCMCs. **P < 0.01 versus vehicle, Dunnett’s test. Typical results with 5 measurements of 3 independent experiments are shown as mean ± SEM. (B and C) To identify O2-sensing molecules on mast cells, TRPA1 was detected by flow cytometry and immunoblotting. Results are representative of 3 independent experiments. A green peak shows a TRPA1-positive reaction and an open peak shows reactivity of a control (B). Instead of first Abs, rabbit serum was used as a control. Strong positive reactions that were visualized around 110 kDa (indicated by an arrow) were estimated as TRPA1 proteins (C). Lane 1, mouse brain membrane; lane 2, C57BL/6 BMCMCs. GAPDH was used as a control for one of the cytosolic endogenous proteins. (D) TRPA1 inhibitor HC-030031 abrogated abnormal neovascularization in mice treated from P11 to P16. n = 8 in each group. **P < 0.01 versus DMSO-injected control mice, Mann-Whitney U test. (E–H) Mast cell stabilizer cromolyn suppressed retinal neovascularization in WT mice. (E) Schematic of the study design. WT and KitWsh/Wsh pups were given cromolyn or vehicle daily by i.p. injections. Mice were injected with cromolyn from P6 to P16, P6 to P11, or P11 to P16 in group 1, 2, or 3, respectively. Eyes were enucleated on P17. (F) Treatment with cromolyn from P6 to P16 reduced pathological neovascularization in WT mice. (G) Cromolyn did not affect abnormal neovascularization in mice in group 2. (H) Neovascular tufts were reduced in WT mice treated with cromolyn between P11 and P16, as in KitWsh/Wsh mice. n = 9 in each group. **P < 0.01 versus PBS-injected control mice, Mann-Whitney U test (F–H). Results are shown as mean ± SEM of values determined from 3 independent experiments (D, F–H).