Critical Role of Notch-1 in Mechanistic Target of Rapamycin Hyperactivity and Vascular Inflammation in Patients With Takayasu Arteritis
Critical Role of Notch-1 in Mechanistic Target of Rapamycin Hyperactivity and Vascular Inflammation in Patients With Takayasu Arteritis
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DOI：
10.1002/art.42103
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发表时间：
2022-06-02
影响因子：
13.3
通讯作者：
Wen, Zhenke
中科院分区：
文献类型：
--
作者：
Jiang, Wanwan;Sun, Mengyao;Wen, Zhenke
目的 大动脉炎（TA）是大血管炎的一种主要类型，其特征是血管层进行性炎症。在我们最近的研究中，我们发现了雷帕霉素靶点 (mTOR) 过度活跃在 TA 促炎 T 细胞分化中的核心作用。本研究旨在探索 T 细胞固有的 mTOR 过度活跃和 TA 血管炎症的潜在机制，重点关注 Notch-1。方法根据Notch-1、激活的Notch-1和HES-1水平测定Notch-1的表达和活性。我们通过磷酸化核糖体蛋白 S6 的细胞内表达检测了 mTOR 活性。通过检测 Th1 和 Th17 谱系决定转录因子来分析促炎 T 细胞的分化。使用 γ-分泌酶抑制剂 DAPT 评估 Notch-1 的功能，并使用短发夹 RNA (shRNA) 策略进行基因敲除。我们使用人源化 NSG 小鼠嵌合体进行了转化研究，其中使用 TA 患者的免疫细胞诱导人类血管炎。结果 TA患者的CD4+ T细胞表达Notch-1（高），导致mTOR过度活跃以及Th1细胞和Th17细胞的自发分化不良。使用 DAPT 和 Notch-1 shRNA 阻断 Notch-1 可有效消除 mTOR 复合物 1 (mTORC1) 激活和促炎 T 细胞分化。从机制上讲，Notch-1 促进 mTOR 表达，与 mTOR 相互作用，并与 mTOR 的溶酶体定位相关。因此，全身施用 DAPT 和 CD4+ T 细胞特异性基因敲除 Notch-1 可以减轻人源化 TA 嵌合体中的血管炎症。结论 TA 患者 CD4+ T 细胞中 Notch-1 表达升高，导致 mTORC1 过度活跃和促炎 T 细胞分化。靶向 Notch-1 是 TA 临床管理的一种有前景的治疗策略。
Objective Takayasu arteritis (TA) is a major type of large vessel vasculitis characterized by progressive inflammation in vascular layers. In our recent study we identified a central role of mechanistic target of rapamycin (mTOR) hyperactivity in proinflammatory T cell differentiation in TA. This study was undertaken to explore potential mechanisms underpinning T cell-intrinsic mTOR hyperactivity and vascular inflammation in TA, with a focus on Notch-1. Methods Notch-1 expression and activity was determined according to Notch-1, activated Notch-1, and HES-1 levels. We detected mTOR activity with intracellular expression of phosphorylated ribosomal protein S6. Differentiation of proinflammatory T cells was analyzed by detecting Th1 and Th17 lineage-determining transcription factors. The function of Notch-1 was evaluated using gamma-secretase inhibitor DAPT and gene knockdown using a short hairpin RNA (shRNA) strategy. We performed our translational study using humanized NSG mouse chimeras in which human vasculitis was induced using immune cells from TA patients. Results CD4+ T cells from TA patients exerted Notch-1(high), leading to mTOR hyperactivity and spontaneous maldifferentiation of Th1 cells and Th17 cells. Blockade of Notch-1 using DAPT and Notch-1 shRNA efficiently abrogated mTOR complex 1 (mTORC1) activation and proinflammatory T cell differentiation. Mechanistically, Notch-1 promoted mTOR expression, interacted with mTOR, and was associated with lysosomal localization of mTOR. Accordingly, systemic administration of DAPT and CD4+ T cell-specific gene knockdown of Notch-1 could alleviate vascular inflammation in humanized TA chimeras. Conclusion Expression of Notch-1 is elevated in CD4+ T cells from TA patients, resulting in mTORC1 hyperactivity and proinflammatory T cell differentiation. Targeting Notch-1 is a promising therapeutic strategy for the clinical management of TA.