Oral Delivery of Bioencapsulated Proteins Across Blood-Brain and Blood-Retinal Barriers
Oral Delivery of Bioencapsulated Proteins Across Blood-Brain and Blood-Retinal Barriers
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DOI：
10.1038/mt.2013.273
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发表时间：
2014-03-01
影响因子：
12.4
通讯作者：
Daniell, Henry
中科院分区：
文献类型：
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作者：
Kohli, Neha;Westerveld, Donevan R.;Daniell, Henry
向与大脑相关的疾病提供神经疗法是一个重大挑战。因此，我们研究了绿色荧光蛋白(GFP)或髓鞘碱性蛋白(MBP)与跨粘膜载体霍乱毒素B亚单位(CTB)融合的口服递送，该亚单位在叶绿体(生物包裹在植物细胞内)中表达，跨越血脑屏障(BBB)和血视网膜屏障(BRB)进入三重转基因阿尔茨海默病(3xTgAD)小鼠的脑和视网膜。人神经母细胞瘤细胞在与CTB-GFP孵育时内化GFP，但不单独与GFP孵育。口服CTB-MBP在健康和3xTgAD小鼠中显示，大脑不同区域的MBP水平增加，跨越完整的血脑屏障。CTB-MBP与人AD和3xTgAD小鼠脑切片体外孵育后，硫代黄素S染色的淀粉样斑块强度最高可降低60%。在体内，服用生物胶囊CTB-MBP的3xTgAD小鼠的海马区和皮质脑区的淀粉样蛋白负荷量降低了70%，同时不溶性淀粉样β42(Aβ42)与可溶部分的比率也降低了。CTB-MBP口服给药可减少Aβ(42)在3xTgAD小鼠视网膜中的蓄积，并防止视网膜神经节细胞的丢失。叶子的冷冻干燥使CTB-MBP浓度增加了17倍，并在胶囊中的长期储存期间保持稳定，有助于低成本地口服跨血脑屏障和BRB的治疗性蛋白质。
Delivering neurotherapeutics to target brain-associated diseases is a major challenge. Therefore, we investigated oral delivery of green fluorescence protein (GFP) or myelin basic protein (MBP) fused with the transmucosal carrier cholera toxin B subunit (CTB), expressed in chloroplasts (bioencapsulated within plant cells) to the brain and retinae of triple transgenic Alzheimer's disease (3xTgAD) mice, across the blood-brain barriers (BBB) and blood-retinal barriers (BRB). Human neuroblastoma cells internalized GFP when incubated with CTB-GFP but not with GFP alone. Oral delivery of CTB-MBP in healthy and 3xTgAD mice shows increased MBP levels in different regions of the brain, crossing intact BBB. Thioflavin S-stained amyloid plaque intensity was reduced up to 60% by CTB-MBP incubation with human AD and 3xTgAD mice brain sections ex vivo. Amyloid loads were reduced in vivo by 70% in hippocampus and cortex brain regions of 3xTgAD mice fed with bioencapsulated CTB-MBP, along with reduction in the ratio of insoluble amyloid beta 42 (A beta(42)) to soluble fractions. CTB-MBP oral delivery reduced A beta(42) accumulation in retinae and prevented loss of retinal ganglion cells in 3xTgAD mice. Lyophilization of leaves increased CTB-MBP concentration by 17-fold and stabilized it during long-term storage in capsules, facilitating low-cost oral delivery of therapeutic proteins across the BBB and BRB.