Intranasally administered extracellular vesicles from human induced pluripotent stem cell-derived neural stem cells quickly incorporate into neurons and microglia in 5xFAD mice.
Intranasally administered extracellular vesicles from human induced pluripotent stem cell-derived neural stem cells quickly incorporate into neurons and microglia in 5xFAD mice.
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DOI：
10.3389/fnagi.2023.1200445
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发表时间：
2023
影响因子：
4.8
通讯作者：
中科院分区：
文献类型：
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作者：
人诱导多能干细胞 (hiPSC) 衍生的神经干细胞 (NSC) 释放的细胞外囊泡 (EV) 由于其负载中的治疗性 miRNA 和蛋白质而具有强大的抗炎和神经源性特性。因此，hiPSC-NSC-EVs 可能是治疗神经退行性疾病（包括阿尔茨海默病（AD））的优秀生物制剂。这项研究调查了鼻内 (IN) 施用的 hiPSC-NSC-EV 是否会快速靶向 3 个月大的 5xFAD 小鼠（β-淀粉样变性和家族性 AD 模型）的前脑、中脑和后脑区域中的各种神经细胞类型。我们给予单剂量的 25 × 109 hiPSC-NSC-EV 标记的 PKH26，并在给药后 45 分钟或 6 小时对接受 EV 的不同组的初始小鼠和 5xFAD 小鼠实施安乐死。给药后 45 分钟，在幼稚小鼠和 5xFAD 小鼠的前脑、中脑和后脑的几乎所有亚区域都发现了 EV，主要靶向并内化为神经元、中间神经元和小胶质细胞，包括 5xFAD 小鼠中的斑块相关小胶质细胞。 EV 还与星形胶质细胞过程的质膜和白质区域的少突胶质细胞体接触。使用神经元标记物对 CD63/CD81 表达的评估证实，神经元内发现的 PKH26 + 颗粒被注射到 hiPSC-NSC-EV 中。给药后 6 小时，EV 在两组所有细胞类型中持续存在，其分布与给药后 45 分钟观察到的分布基本匹配。面积分数 (AF) 分析显示，在幼稚小鼠和 5xFAD 小鼠中，在两个时间点都有较高比例的 EV 融入前脑区域。然而，在 IN 给药后 45 分钟，5xFAD 小鼠中前脑区域细胞层内以及中脑和后脑区域小胶质细胞内 EV 的 AF 低于初始小鼠，这意味着淀粉样变性降低了 EV 外显率。总的来说，这些结果提供了新的证据，证明治疗性 hiPSC-NSC-EV 的 IN 给药是在淀粉样变性早期阶段将此类 EV 引导至所有大脑区域的神经元和神经胶质细胞的有效途径。由于在多个大脑区域观察到 AD 的病理变化，因此在淀粉样变性早期阶段将治疗性 EV 递送到几乎每个大脑区域的各种神经细胞对于促进神经保护和抗炎作用具有吸引力。
Extracellular vesicles (EVs) released by human-induced pluripotent stem cell (hiPSC)-derived neural stem cells (NSCs) have robust antiinflammatory and neurogenic properties due to therapeutic miRNAs and proteins in their cargo. Hence, hiPSC-NSC-EVs are potentially an excellent biologic for treating neurodegenerative disorders, including Alzheimer’s disease (AD). This study investigated whether intranasally (IN) administered hiPSC-NSC-EVs would quickly target various neural cell types in the forebrain, midbrain, and hindbrain regions of 3-month-old 5xFAD mice, a model of β-amyloidosis and familial AD. We administered a single dose of 25 × 109 hiPSC-NSC-EVs labeled with PKH26, and different cohorts of naïve and 5xFAD mice receiving EVs were euthanized at 45 min or 6 h post-administration. At 45 min post-administration, EVs were found in virtually all subregions of the forebrain, midbrain, and hindbrain of naïve and 5xFAD mice, with predominant targeting and internalization into neurons, interneurons, and microglia, including plaque-associated microglia in 5xFAD mice. EVs also came in contact with the plasma membranes of astrocytic processes and the soma of oligodendrocytes in white matter regions. Evaluation of CD63/CD81 expression with the neuronal marker confirmed that PKH26 + particles found within neurons were IN administered hiPSC-NSC-EVs. At 6 h post-administration, EVs persisted in all cell types in both groups, with the distribution mostly matching what was observed at 45 min post-administration. Area fraction (AF) analysis revealed that, in both naïve and 5xFAD mice, higher fractions of EVs incorporate into forebrain regions at both time points. However, at 45 min post-IN administration, AFs of EVs within cell layers in forebrain regions and within microglia in midbrain and hindbrain regions were lower in 5xFAD mice than naïve mice, implying that amyloidosis reduces EV penetrance. Collectively, the results provide novel evidence that IN administration of therapeutic hiPSC-NSC-EVs is an efficient avenue for directing such EVs into neurons and glia in all brain regions in the early stage of amyloidosis. As pathological changes in AD are observed in multiple brain areas, the ability to deliver therapeutic EVs into various neural cells in virtually every brain region in the early stage of amyloidosis is attractive for promoting neuroprotective and antiinflammatory effects.
影响因子：
4.7
作者：
Pauwels MJ;Vandendriessche C;Vandenbroucke RE
通讯作者：
Vandenbroucke RE