用于靶向治疗小鼠视神经脊髓炎谱系障碍的铁蛋白ApoE纳米载体
Ferritin-ApoE nanocarrier for targeted therapy of neuromyelitis optica spectrum disorder in mice.
摘要(中文译文)
视神经脊髓炎谱系疾病（NMOSD）是一种累及中枢神经系统（CNS）的慢性炎症性自身免疫性疾病，以抗水通道蛋白4（AQP4）抗体介导的星形胶质细胞损伤为特征，进而导致脱髓鞘。我们前期研究发现，载脂蛋白E（ApoE）肽通过促进星形胶质细胞-小胶质细胞间通讯在NMOSD小鼠中发挥保护作用。然而，由于全身给药时血脑屏障（BBB）穿透能力有限，其治疗潜力受到限制。在此，我们设计了一种基于重链铁蛋白（HFn）的纳米载体，负载ApoE肽（HFn-ApoE），专门针对中枢神经系统递送进行工程化改造。HFn-ApoE通过基因工程构建，将HFn编码序列与ApoE肽编码序列融合于重组质粒中。通过向脑内经颅共注射AQP4-IgG和人补体（hC）诱导急性NMOSD小鼠模型。静脉注射后，利用活体荧光成像追踪Cy5.5标记的HFn-ApoE的分布，以确认其在脑及外周器官中的存在。采用T2加权7特斯拉磁共振成像（7T-MRI）对脑内病灶进行定量。通过脑切片免疫染色评估NMOSD的神经病理特征。通过流式细胞术分析神经炎症和免疫细胞浸润。通过Western blot（WB）分析研究HFn-ApoE调控的关键信号通路。通过免疫共沉淀验证HFn-ApoE与其受体的相互作用，并使用邻位连接 assay（PLA）在小胶质细胞上可视化。最后，采用Morris水迷宫（MWM）实验评估其对空间学习和记忆的治疗效果。HFn-ApoE有效穿越血脑屏障，减轻NMOSD小鼠的病灶进展和脱髓鞘，并保护AQP4表达和星形胶质细胞完整性。该治疗诱导星形胶质细胞反应的空间和表型重塑，显著减少病灶周围星形胶质细胞的过度聚集，同时促进增殖和修复表型。此外，HFn-ApoE促进小胶质细胞向抗炎状态极化，减少外周免疫细胞浸润。机制上，HFn-ApoE通过小胶质细胞中的低密度脂蛋白受体相关蛋白1（LRP1）-核因子κB（NF-κB）信号轴发挥抗炎作用。HFn-ApoE与LRP1的功能性结合抑制NF-κB抑制蛋白（IκBα）磷酸化，从而抑制NF-κB核转位及随后促炎细胞因子的释放，包括白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）。敲低LRP1可逆转这些效应，凸显了LRP1-NF-κB信号轴在该纳米治疗药物疗效中的重要性。HFn-ApoE治疗改善了NMOSD小鼠的空间学习能力并挽救了记忆缺陷。本研究表明，工程化纳米药物HFn-ApoE通过增强血脑屏障穿透并通过LRP1-NF-κB信号轴抑制神经炎症，是缓解NMOSD病理的有前景的靶向治疗手段。
Abstract
Neuromyelitis optica spectrum disorder (NMOSD) is a chronic inflammatory autoimmune disease affecting the central nervous system (CNS), characterized by anti-aquaporin 4 (AQP4) antibody-mediated damage to astrocytes, resulting in subsequent demyelination. Our prior work identified the protective effects of the apolipoprotein E (ApoE) peptide in NMOSD mice by promoting astrocyte-microglia intercellular communication. However, its therapeutic potential is restricted due to the limited penetration of the blood-brain barrier (BBB) with systemic administration. Here, we designed a heavy-chain ferritin (HFn)-based nanocarrier containing the ApoE peptide (HFn-ApoE), specifically engineered for CNS delivery. HFn-ApoE was constructed through genetic engineering by fusing the coding sequence of HFn with that of the ApoE peptide in a recombinant plasmid. An acute NMOSD mouse model was induced by transcranial co-injection of AQP4-IgG and human complement (hC) into the brain. The distribution of Cy5.5-labeled HFn-ApoE post intravenous injection was tracked using in vivo fluorescence imaging to confirm its presence in the brain and peripheral organs. Lesions in the brain were quantified using T2-weighted 7 Tesla magnetic resonance imaging (7T-MRI). Neuropathological features of NMOSD were evaluated by immunostaining of brain sections. Neuroinflammation and immune cell infiltration were analyzed via flow cytometry. The key signaling pathways regulated by HFn-ApoE were investigated through Western blot (WB) analysis. The interaction between HFn-ApoE and its receptors was validated through co-immunoprecipitation and visualized on microglia using proximity ligation assay (PLA). Finally, the therapeutic effect on spatial learning and memory was evaluated using the Morris water maze (MWM) test. The HFn-ApoE effectively crossed the BBB, attenuated lesion progression and demyelination, as well as preserved AQP4 expression and astrocytic integrity in NMOSD mice. The treatment induced a spatial and phenotypic restructuring of the astrocytic response, notably reducing excessive astrocyte accumulation around lesions while encouraging a proliferative and reparative phenotype. Furthermore, HFn-ApoE influenced microglial polarization towards an anti-inflammatory state, reducing infiltration of peripheral immune cells. Mechanistically, HFn-ApoE exerted its anti-inflammatory effects through the low-density lipoprotein receptor-related protein 1 (LRP1) -nuclear factor kappa B (NF-κB) signaling axis in microglia. Functional binding of HFn-ApoE to LRP1 suppressed inhibitor of NF-κB (IκBα) phosphorylation, thereby inhibiting NF-κB nuclear translocation and the subsequent release of pro-inflammatory cytokines, including interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Knocking down LRP1 reversed these effects, highlighting the importance of the LRP1-NF-κB signaling axis in the nanotherapeutic's efficacy. Treatment with HFn-ApoE improved spatial learning and rescued memory deficits in NMOSD mice. This study demonstrates that the engineered nanodrug HFn-ApoE is a promising targeted therapy for alleviating NMOSD pathology by enhancing BBB penetration and suppressing neuroinflammation through the LRP1-NF-κB signaling axis.
如何引用
AMA
Xin Zhao, Qian Liang, Kangqi Lin, Shihe Jiang, Tian Yang, Mengjiao Sun, et al. Ferritin-ApoE nanocarrier for targeted therapy of neuromyelitis optica spectrum disorder in mice.. Journal of neuroinflammation. 2026; doi:10.1186/s12974-026-03935-8.
APA
Xin Zhao, Qian Liang, Kangqi Lin, Shihe Jiang, Tian Yang, Mengjiao Sun, et al (2026). Ferritin-ApoE nanocarrier for targeted therapy of neuromyelitis optica spectrum disorder in mice.. Journal of neuroinflammation. https://doi.org/10.1186/s12974-026-03935-8
GB/T 7714
Xin Zhao, Qian Liang, Kangqi Lin, Shihe Jiang, Tian Yang, Mengjiao Sun, et al. Ferritin-ApoE nanocarrier for targeted therapy of neuromyelitis optica spectrum disorder in mice.[J]. Journal of neuroinflammation, 2026 doi:10.1186/s12974-026-03935-8.
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