Ibrutinib增强巨噬细胞清除感染结核菌的作用和机制

How to improve tuberculosis, especially multi-drug resistant tuberculosis therapy efficacy is a grand project at present. Our previous study showed that BTK target molecule ibrutinib could significantly enhance mycobacterium tuberculosis clearance in macrophage, further studies showed that ibrutinib does-dependly induced ROS reaction in macrophage, but didn't promote apoptosis, NO production and autophage in Mtb infected macrophage. Based on our previous results, we hypothesize that ibrutinib enhances clearance of TB in macrophage by inducing high level of ROS reaction. To clarify the hypothesis, we are planning to clarify the enchanced clearance of drug-resistant Mtb in macrophage induced with ibrutinib stimulation; to identify the key factor of enhancing infected Mtb clearance in macrophage with ibrutinib stimulation, to clarify the regulation mechanism of ibrutinib enhancing ROS production in macrophage. Finally, we ought to complete the hypothesis via using Tn-seq to clarify the pressure on Mtb in macrophage with ibrutinib stimulation. The result of this study will provide new clues for tuberculosis, especially drug-resistant tuberculosis therapy.

如何提高结核病尤其是耐多药结核的治疗效果是当前重要课题。我们前期发现BTK靶向药Ibrutinib显著增强巨噬细胞清除感染结核菌，进一步研究显示Ibrutinib以浓度依赖的方式显著增加巨噬细胞中活性氧自由基（ROS）水平，而不促进结核菌感染巨噬细胞的凋亡、一氧化氮生成和自噬作用。基于前期结果，我们提出Ibrutinib通过上调巨噬细胞中ROS水平增强清除感染结核菌的假说。本研究拟在前期实验基础上进一步明确Ibrutinib增强巨噬细胞清除感染耐药结核菌的作用；鉴定Ibrutinib促进巨噬细胞ROS生成增强清除感染结核菌的关键因素，确定Ibrutinib促进巨噬细胞ROS生成的关键通路靶点；进而通过转座子技术研究Ibrutinib对胞内结核菌的选择压力，明确Ibrutinib增强巨噬细胞清除感染结核菌的机制。本项目的顺利开展将为结核病尤其是耐药结核治疗提供新的思路。

背景：如何提高结核病尤其是耐多药结核的治疗效果是当前重要课题。我们前期发现BTK靶向药Ibrutinib显著增强巨噬细胞清除感染结核菌，进一步研究显示Ibrutinib以浓度依赖的方式显著增加巨噬细胞中活性氧自由基（ROS）水平，而不促进结核菌感染巨噬细胞的凋亡、一氧化氮生成和自噬作用。基于前期结果，我们提出Ibrutinib通过上调巨噬细胞中ROS水平增强清除感染结核菌的假说。.主要研究内容、重要结果、关键数据：本研究在前期基础上进一步研究了Ibrutinib增强巨噬细胞清除感染有毒结核菌株的作用；修正了科学假说，鉴定出Ibrutinib促进巨噬细胞杀菌功能是通过自噬作用起效；确定Ibrutinib诱导巨噬细胞自噬是通过BTK/AKT/mTOR通路；最后在结核病小鼠模型验证出Ibrutinib可显著降低脾脏和肺门淋巴结中结核菌载量。.科学意义：本研究完成了Ibrutinib增强巨噬细胞杀灭胞内结核菌的作用和机制，达到预期研究目标，同时评估了Ibrutinib具有体内治疗结核病的潜力，为结核病的辅助治疗提供了候选药物。