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.

中文翻译：

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GATA3 的远端增强子调节 Th2 分化和过敏性炎症


哮喘是一种广泛存在的气道疾病，其中依赖 GATA3 的 2 型辅助 T (Th2) 细胞和 2 类先天淋巴细胞 (ILC2) 发挥着至关重要的作用。哮喘相关单核苷酸多态性 (SNP) 富集于 10p14 (hG900) 中 GATA3 下游 926-970 kb 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。接下来，我们通过 CRISPR-Cas9 系统生成了缺乏 mG900 区域的小鼠（mG900KO 小鼠），并在稳态条件和木瓜蛋白酶或诱导的过敏性气道炎症下分析了 mG900KO 小鼠中辅助 T 细胞和 ILC 的发育和功能。屋尘螨 (HDM)。 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过敏性炎症和 Th2 分化减少。通过环状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表明，mG900 区域与 Gata3 的转录起始位点相互作用，从而影响 Th2 细胞中的染色质构象。 这些发现表明 mG900 区域在 Th2 分化中发挥关键作用，从而增强过敏性气道炎症。