Hao Hongying, Yin Tingyu, Li Tuo, Zhou Xu, Ren Honglei, Liu Mingming, Huang Huachen, Qi Caiyun, Xiu Yuwen, Qiu Wenjin, Wang Danni, Shi Mengxuan, Wang Xiaoying, Dumont Aaron S, Liu Qiang
Department of Neurology, Tianjin Neurological Institute, Tianjin Institute of Immunology, State Key Laboratory of Experimental Hematology, International Joint Laboratory of Ocular Diseases, Ministry of Education, Haihe Laboratory of Cell Ecosystem, Laboratory of Post-Neuroinjury Neurorepair and Regeneration in Central Nervous System Tianjin & Ministry of Education, Tianjin Medical University General Hospital, Tianjin 300052, China.
Department of Neurosurgery, Tianjin Huanhu Hospital, Tianjin 300350, China.
Theranostics. 2025 Jan 1;15(2):494-508. doi: 10.7150/thno.101024. eCollection 2025.
Intracerebral hemorrhage (ICH) is a devastating form of stroke with a lack of effective treatments. Following disease onset, ICH activates microglia and recruits peripheral leukocytes into the perihematomal region to amplify neural injury. Bruton's tyrosine kinase (BTK) controls the proliferation and survival of various myeloid cells and lymphocytes. However, the role of BTK in neuroinflammation and ICH injury remains poorly understood. Peripheral blood samples were collected from ICH patients and healthy controls to measure BTK expression profile in immune cell subsets. C57BL/6 mice were used to measure BTK expression and the activity of immune cell subsets following ICH induction. Neurological tests, brain water content, Evans blue leakage, MRI were used to assess the therapeutic effects of ibrutinib on ICH injury. Flow cytometry was used to investigate immune cell infiltration and microglial activity. Microglia were depleted using a CSF1R inhibitor PLX5622. Gr-1 myeloid cells and B cells were depleted using monoclonal antibodies. Microglia-like BV2 cells were cultured to test the effects of BTK inhibition on these cells. In humans and mice, we found that BTK was remarkably upregulated in myeloid cells after ICH. Inhibition of BTK using ibrutinib led to reduced neurological deficits, perihematomal edema, brain water content and blood-brain barrier disruption. BTK inhibition suppressed the inflammatory activity of microglia and brain infiltration of leukocytes. In contrast, BTK inhibition did not alter the counts of peripheral immune cells other than B cells. Further, the depletion of microglia or Gr-1 myeloid cells ablated the protective effects of BTK inhibition against ICH injury. Notably, the depletion of B cells did not alter the protective effects of BTK inhibition against ICH injury. This suggests that the benefit of BTK inhibition in ICH mainly involves its impact on microglia and Gr-1 myeloid cells. Our findings demonstrate that BTK inhibition attenuates neuroinflammation and ICH injury, which warrants further investigation as a potential therapy for ICH.
脑出血（ICH）是一种具有毁灭性的中风形式，缺乏有效的治疗方法。疾病发作后，ICH会激活小胶质细胞并将外周白细胞募集到血肿周围区域，从而加剧神经损伤。布鲁顿酪氨酸激酶（BTK）控制各种髓样细胞和淋巴细胞的增殖与存活。然而，BTK在神经炎症和ICH损伤中的作用仍知之甚少。收集ICH患者和健康对照者的外周血样本，以测量免疫细胞亚群中的BTK表达谱。使用C57BL/6小鼠测量ICH诱导后BTK的表达以及免疫细胞亚群的活性。通过神经学测试、脑含水量、伊文思蓝渗漏、磁共振成像来评估依鲁替尼对ICH损伤的治疗效果。采用流式细胞术研究免疫细胞浸润和小胶质细胞活性。使用集落刺激因子1受体（CSF1R）抑制剂PLX5622清除小胶质细胞。使用单克隆抗体清除Gr-1髓样细胞和B细胞。培养小胶质细胞样BV2细胞，以测试BTK抑制对这些细胞的影响。在人和小鼠中，我们发现ICH后髓样细胞中的BTK显著上调。使用依鲁替尼抑制BTK可减少神经功能缺损、血肿周围水肿、脑含水量和血脑屏障破坏。BTK抑制可抑制小胶质细胞的炎症活性和白细胞向脑内的浸润。相比之下，BTK抑制除了B细胞外，不会改变外周免疫细胞的数量。此外，清除小胶质细胞或Gr-1髓样细胞会消除BTK抑制对ICH损伤的保护作用。值得注意的是，清除B细胞不会改变BTK抑制对ICH损伤的保护作用。这表明BTK抑制在ICH中的益处主要涉及其对小胶质细胞和Gr-1髓样细胞的影响。我们的研究结果表明，BTK抑制可减轻神经炎症和ICH损伤，作为ICH的一种潜在治疗方法，值得进一步研究。
