己糖激酶HK3通过代谢重编程调控巨噬细胞极化参与自身免疫甲状腺疾病发生的研究

Autoimmune thyroid diseases (AITD), the most common autoimmune disease, mainly include Graves' disease (GD) and Hashimoto's thyroiditis (HT); loss of immune tolerance caused by immune imbalance plays a critical role in the pathogenesis of AITD. Previous studies have found that macrophage polarization is intensively involved in the pathological process of multiple autoimmune diseases, but its role in AITD still remains elusive. Our preliminary research found that the glycolysis key enzyme hexokinase 3 (HK3) is aberrantly over-expressed in many autoimmune diseases including AITD. Further preliminary study indicated that HK3 might regulate macrophage polarization through metabolic reprogramming. Hereby we hypothesize that macrophage polarization has an important role in the pathogenesis of AITD, and HK3 exerts a critical role in the pathophysiology of AITD through regulating macrophage polarization by metabolic reprogramming. To test this hypothesis, we will perform clinical research, in-vitro cell culture experiments and in-vivo experiments with AITD animal models by using a variety of techniques, such as CRISPR/Cas9 method, RNA interference, high-throughput sequencing and molecular immunology, to study the regulatory role of HK3 in macrophage polarization and its relevant molecular mechanisms and to investigate their roles in the pathogenesis of AITD, which will provide a theoretical foundation for novel targeted immunotherapy of AITD.

自身免疫甲状腺疾病(AITD)是最常见的自身免疫性疾病，主要包括Graves病和桥本甲状腺炎。免疫稳态失衡引发的免疫耐受终止在其发病机制中起关键作用。研究显示巨噬细胞极化失衡与多种自身免疫性疾病的发生密切相关，但其在AITD中的作用尚不明确。我们前期研究发现作为糖酵解关键酶的己糖激酶3(HK3)在包括AITD在内的多种自身免疫疾病中的表达水平异常升高，并且HK3可能与巨噬细胞极化有关。因此我们提出以下假说，巨噬细胞极化在AITD发病机制中有重要作用，而HK3可以通过代谢重编程调控巨噬细胞极化并参与AITD的发生。为了验证这一假说，我们拟采用临床研究、体外细胞培养和动物疾病模型，综合运用CRISPR/Cas9技术、RNA干扰、高通量测序和分子免疫学等实验方法研究HK3对巨噬细胞极化的影响及可能的调控机制，研究其在AITD发病机制中的作用，为探索AITD免疫靶向治疗策略奠定理论基础。

自身免疫性甲状腺疾病(AITD)是最常见的自身免疫性疾病，包括Graves病(GD)和桥本甲状腺炎(HT)。目前，AITD的发病机制尚不完全清楚。我们的研究旨在检测AITD患者中巨噬细胞极化不平衡的存在，探讨高碘是否会导致巨噬细胞极化不平衡，并探讨代谢重编程关键基因在高碘引起的巨噬细胞极化不平衡中的作用。我们协同使用了各种研究策略，如系统生物学、临床研究、细胞培养和小鼠疾病模型。.实验结果证实：.（1）基因集富集分析(GSEA)显示M1型巨噬细胞超极化参与了AITD的发病机制。.（2）体外和体内实验表明，高碘可影响M1型或M2型巨噬细胞的极化及其相关细胞因子。RRA方法显示己糖激酶3 (HK3)是自身免疫性疾病中异常表达最显著的代谢基因。体外和体内研究表明，HK3可介导高碘诱导的巨噬细胞极化。.（3）单核巨噬细胞特异表面标记分子 CD14 与AITD的发病风险存在遗传关联。.综上所述，M1型巨噬细胞的超极化与AITD的发病密切相关；高碘可增加巨噬细胞中HK3的表达，促进巨噬细胞向M1极化；而靶向HK3可抑制高碘诱导的M1极化。