GATA3的远端增强子调节Th2分化和过敏性炎症
2024/07/30
摘要
哮喘是一种广泛存在的气道疾病，其中GATA3依赖性2型辅助性T细胞（Th2）和2组固有淋巴细胞（ILC2）发挥着至关重要的作用。哮喘相关单核苷酸多态性（SNPs）富集于10p14（hG900）中GATA3下游926-970kb的区域。然而，hG900如何影响过敏性气道炎症的发病机制尚不清楚。为了研究哮喘相关的GATA3增强子区域在实验性过敏性气道炎症中的作用，我们首先通过流式细胞术和ChIP-qPCR分析了GATA3表达与hG900区域激活之间的相关性。我们发现hG900区域增强子的激活与人外周T细胞亚群中GATA3的水平密切相关。我们接下来生成的缺乏mG900区域的小鼠（mG900KO小鼠）由CRISPR-Cas9系统生成，并在稳态条件下分析mG900KO小鼠中辅助T细胞和ILCs的发育和功能，以及木瓜蛋白酶或屋尘螨（HDM）诱导的过敏性气道炎症。mG900的缺失不影响稳态条件下淋巴细胞的发育或木瓜蛋白酶诱导的过敏性气道炎症。然而，mG900KO小鼠在HDM诱导的过敏性气道炎症中表现出减少的过敏性炎症和Th2分化。通过环形染色体构象捕获结合高通量测序（4C-seq）对Gata3周围的染色质构象进行分析，发现mG900区域与Gata3的转录起始位点相互作用，影响Th2细胞中的染色质构型。这些发现表明mG900区域在Th2分化中起着关键作用，从而增强过敏性气道炎症。
A distal enhancer of GATA3 regulates Th2 differentiation and allergic inflammation
Takashi Kumagai, Arifumi Iwata, Hiroki Furuya, Kodai Kato, Atsushi Okabe, Yosuke Toda, Mizuki Kanai, Lisa Fujimura, Akemi Sakamoto, Takahiro Kageyama, Shigeru Tanaka, Akira Suto, Masahiko Hatano, Atsushi Kaneda, Hiroshi Nakajima
Abstract
Asthma is a widespread airway disorder where GATA3-dependent Type-2 helper T (Th2) cells and group 2 innate lymphoid cells (ILC2s) play vital roles. Asthma-associated single nucleotide polymorphisms (SNPs) are enriched in a region located 926-970 kb downstream from GATA3 in the 10p14 (hG900). However, it is unknown how hG900 affects the pathogenesis of allergic airway inflammation. To investigate the roles of the asthma-associated GATA3 enhancer region in experimental allergic airway inflammation, we first examined the correlation between GATA3 expression and the activation of the hG900 region was analyzed by flow cytometry and ChIP-qPCR. We found that The activation of enhancers in the hG900 region was strongly correlated to the levels of GATA3 in human peripheral T cell subsets. We next generated mice lacking the mG900 region (mG900KO mice) were generated by the CRISPR-Cas9 system, and the development and function of helper T cells and ILCs in mG900KO mice were analyzed in steady-state conditions and allergic airway inflammation induced by papain or house dust mite (HDM). The deletion of the mG900 did not affect the development of lymphocytes in steady-state conditions or allergic airway inflammation induced by papain. However, mG900KO mice exhibited reduced allergic inflammation and Th2 differentiation in the HDM-induced allergic airway inflammation. The analysis of the chromatin conformation around Gata3 by circular chromosome conformation capture coupled to high-throughput sequencing (4C-seq) revealed that the mG900 region interacted with the transcription start site of Gata3 with an influencing chromatin conformation in Th2 cells. These findings indicate that the mG900 region plays a pivotal role in Th2 differentiation and thus enhances allergic airway inflammation.
哮喘是一种广泛存在的气道疾病，其中GATA3依赖性2型辅助性T细胞（Th2）和2组固有淋巴细胞（ILC2）发挥着至关重要的作用。哮喘相关单核苷酸多态性（SNPs）富集于10p14（hG900）中GATA3下游926-970kb的区域。然而，hG900如何影响过敏性气道炎症的发病机制尚不清楚。为了研究哮喘相关的GATA3增强子区域在实验性过敏性气道炎症中的作用，我们首先通过流式细胞术和ChIP-qPCR分析了GATA3表达与hG900区域激活之间的相关性。我们发现hG900区域增强子的激活与人外周T细胞亚群中GATA3的水平密切相关。我们接下来生成的缺乏mG900区域的小鼠（mG900KO小鼠）由CRISPR-Cas9系统生成，并在稳态条件下分析mG900KO小鼠中辅助T细胞和ILCs的发育和功能，以及木瓜蛋白酶或屋尘螨（HDM）诱导的过敏性气道炎症。mG900的缺失不影响稳态条件下淋巴细胞的发育或木瓜蛋白酶诱导的过敏性气道炎症。然而，mG900KO小鼠在HDM诱导的过敏性气道炎症中表现出减少的过敏性炎症和Th2分化。通过环形染色体构象捕获结合高通量测序（4C-seq）对Gata3周围的染色质构象进行分析，发现mG900区域与Gata3的转录起始位点相互作用，影响Th2细胞中的染色质构型。这些发现表明mG900区域在Th2分化中起着关键作用，从而增强过敏性气道炎症。
(中日友好医院呼吸与危重症医学科 李红雯 摘译 林江涛 审校)
(Proc Natl Acad Sci U S A. 2024 Jul 2;121(27):e2320727121. doi: 10.1073/pnas.2320727121.)
(Proc Natl Acad Sci U S A. 2024 Jul 2;121(27):e2320727121. doi: 10.1073/pnas.2320727121.)
A distal enhancer of GATA3 regulates Th2 differentiation and allergic inflammation
Takashi Kumagai, Arifumi Iwata, Hiroki Furuya, Kodai Kato, Atsushi Okabe, Yosuke Toda, Mizuki Kanai, Lisa Fujimura, Akemi Sakamoto, Takahiro Kageyama, Shigeru Tanaka, Akira Suto, Masahiko Hatano, Atsushi Kaneda, Hiroshi Nakajima
Abstract
Asthma is a widespread airway disorder where GATA3-dependent Type-2 helper T (Th2) cells and group 2 innate lymphoid cells (ILC2s) play vital roles. Asthma-associated single nucleotide polymorphisms (SNPs) are enriched in a region located 926-970 kb downstream from GATA3 in the 10p14 (hG900). However, it is unknown how hG900 affects the pathogenesis of allergic airway inflammation. To investigate the roles of the asthma-associated GATA3 enhancer region in experimental allergic airway inflammation, we first examined the correlation between GATA3 expression and the activation of the hG900 region was analyzed by flow cytometry and ChIP-qPCR. We found that The activation of enhancers in the hG900 region was strongly correlated to the levels of GATA3 in human peripheral T cell subsets. We next generated mice lacking the mG900 region (mG900KO mice) were generated by the CRISPR-Cas9 system, and the development and function of helper T cells and ILCs in mG900KO mice were analyzed in steady-state conditions and allergic airway inflammation induced by papain or house dust mite (HDM). The deletion of the mG900 did not affect the development of lymphocytes in steady-state conditions or allergic airway inflammation induced by papain. However, mG900KO mice exhibited reduced allergic inflammation and Th2 differentiation in the HDM-induced allergic airway inflammation. The analysis of the chromatin conformation around Gata3 by circular chromosome conformation capture coupled to high-throughput sequencing (4C-seq) revealed that the mG900 region interacted with the transcription start site of Gata3 with an influencing chromatin conformation in Th2 cells. These findings indicate that the mG900 region plays a pivotal role in Th2 differentiation and thus enhances allergic airway inflammation.
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合并功能性消化不良反映了 IL-33 介导的哮喘气道神经元功能障碍
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高盐饮食对人类和小鼠哮喘的影响：对特定T细胞特征和微生物组的影响
