METTL23 mutation alters histone H3R17 methylation in normal-tension glaucoma
Yang Pan, … , Toru Nakazawa, Takeshi Iwata
Yang Pan, … , Toru Nakazawa, Takeshi Iwata
Published September 13, 2022
Citation Information: J Clin Invest. 2022;132(21):e153589. https://doi.org/10.1172/JCI153589.
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Research Article Ophthalmology

METTL23 mutation alters histone H3R17 methylation in normal-tension glaucoma

Abstract

Normal-tension glaucoma (NTG) is a heterogeneous disease characterized by retinal ganglion cell (RGC) death leading to cupping of the optic nerve head and visual field loss at normal intraocular pressure (IOP). The pathogenesis of NTG remains unclear. Here, we describe a single nucleotide mutation in exon 2 of the methyltransferase-like 23 (METTL23) gene identified in 3 generations of a Japanese family with NTG. This mutation caused METTL23 mRNA aberrant splicing, which abolished normal protein production and altered subcellular localization. Mettl23–knock-in (Mettl23+/G and Mettl23G/G) and -knockout (Mettl23+/– and Mettl23–/–) mice developed a glaucoma phenotype without elevated IOP. METTL23 is a histone arginine methyltransferase expressed in murine and macaque RGCs. However, the novel mutation reduced METTL23 expression in RGCs of Mettl23G/G mice, which recapitulated both clinical and biological phenotypes. Moreover, our findings demonstrated that METTL23 catalyzed the dimethylation of H3R17 in the retina and was required for the transcription of pS2, an estrogen receptor α target gene that was critical for RGC homeostasis through the negative regulation of NF-κB–mediated TNF-α and IL-1β feedback. These findings suggest an etiologic role of METTL23 in NTG with tissue-specific pathology.

Authors

Yang Pan, Akiko Suga, Itaru Kimura, Chojiro Kimura, Yuriko Minegishi, Mao Nakayama, Kazutoshi Yoshitake, Daisuke Iejima, Naoko Minematsu, Megumi Yamamoto, Fumihiko Mabuchi, Mitsuko Takamoto, Yukihiro Shiga, Makoto Araie, Kenji Kashiwagi, Makoto Aihara, Toru Nakazawa, Takeshi Iwata

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Figure 3

The Mettl23 mutation causes exon skipping in vivo.

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The Mettl23 mutation causes exon skipping in vivo. (A) Sequence of the t...
(A) Sequence of the targeted region in Mettl23. Mettl23-KI and -KO mice were produced by genome editing using the CRISPR/Cas9 system. The mutant sequence is boxed in red. The STOP codon is indicated by blue letters. (B) Number of pups born after natural mating of Mettl23-KI or -KO mice. Hetero/Hetero, mating of heterozygous pairs of mice; Homo/Hetero, mating of homozygous and heterozygous mice. Dots represent the number of pups per litter on the day of birth. (C) Body weights of WT (n = 10), Mettl23-KI Mettl23+/G (n = 10), Mettl23G/G (n = 10), Mettl23+/– (n = 10), and Mettl23–/– mice (n = 10). At 2 months of age, all Mettl23 genetic mice had body weights within the normal range. (D) IOPs for WT (n = 14), Mettl23+/G (n = 12), Mettl23G/G (n = 13), Mettl23+/– (n = 12), and Mettl23–/– (n = 12) mice. At 2 months of age, all mice were examined under identical conditions and exhibited IOPs within the normal range. (E) Gel electrophoresis of RT-PCR products from retinas of Mettl23+/G and Mettl23G/G mice. The contents of the application products were determined by TA cloning following Sanger sequencing. All data are presented as the mean ± SEM. *P < 0.05, by Student’s t test.