Characterization of Hit Compounds Identified from High-throughput Screening for their Effect on Blood-brain Barrier Integrity and Amyloid-β Clearance: In Vitro and In Vivo Studies.
Characterization of Hit Compounds Identified from High-throughput Screening for their Effect on Blood-brain Barrier Integrity and Amyloid-β Clearance: In Vitro and In Vivo Studies.
复制标题
DOI：
10.1016/j.neuroscience.2018.03.028
复制
发表时间：
2018-05-21
期刊：
影响因子：
3.3
通讯作者：
Kaddoumi A
中科院分区：
文献类型：
--
作者：
Elfakhri KH;Duong QV;Langley C;Depaula A;Mousa YM;Lebeouf T;Cain C;Kaddoumi A
在阿尔茨海默病（AD）中，血脑屏障（BBB）受损，因此治疗靶向BBB以增强其完整性和功能可能是治疗、减缓或保持AD进展的独特方法。最近，我们已经开发了一种体外高通量筛选试验来筛选增加基于细胞的BBB模型的完整性的化合物。初步筛选的结果鉴定了增强单层完整性的多个命中化合物。在此，对选定的命中化合物，即8-溴鸟苷环一磷酸盐、JW 74、1，10-菲咯啉一水合物、SB 216763和α-生育酚进行了进一步表征。对化合物进行浓度依赖性研究，以确定其EC 50和通过荧光黄渗透性和淀粉样蛋白-β（Aβ）跨单层转运增强基于细胞的模型完整性的效力。这些化合物表现出不同的EC 50，以增强单层完整性，范围为0.4 - 12.8 μM，以及对增强Aβ转运的不同作用，观察到α-生育酚的转运最高（增加2.2倍）。这些作用与紧密连接蛋白（如claudin-5和/或ZO-1）以及Aβ主要转运蛋白LRP 1和P-糖蛋白水平升高相关。在AD小鼠模型中进行了α-生育酚的体内研究;与体外结果一致，α-生育酚显著增加了通过IgG外渗测量的BBB完整性，并降低了脑Aβ水平。总之，研究结果支持我们开发的基于细胞的BBB模型作为选择命中化合物的功能预测体内工具，并表明增强BBB紧密性和功能具有减少与AD相关的Aβ病理的潜力。
In Alzheimer’s disease (AD) the blood-brain barrier (BBB) is compromised, thus therapeutic targeting of the BBB to enhance its integrity and function could be a unique approach to treat, slow or hold the progression of AD. Recently, we have developed an in vitro high-throughput screening assay to screen for compounds that increase the integrity of a cell-based BBB model. Results from primary screen identified multiple hit compounds that enhanced the monolayer integrity. Herein, further characterization of selected hit compounds, namely 8-bromoguanosine cyclic monophosphate, JW74, 1,10-phenanthroline monohydrate, SB216763 and α-tocopherol was performed. Compounds were subjected to concentration-dependent studies to determine their EC50 and potency to enhance the cell-based model integrity by the Lucifer Yellow permeability and amyloid-beta (Aβ) transport across the monolayer. The compounds demonstrated different EC50s to enhance the monolayer integrity ranging from 0.4 to 12.8 μM, and different effect on enhancing Aβ transport with highest transport observed for α-tocopherol (2.2-fold increase). Such effects were associated with increased levels of tight junction proteins such as claudin-5 and/or ZO-1, and Aβ major transport proteins LRP1 and P-glycoprotein. In vivo studies for α-tocopherol were performed in AD mouse model; consistent with the in vitro results α-tocopherol significantly increased BBB integrity measured by IgG extravasation, and reduced brain Aβ levels. In conclusion, findings support our developed cell-based BBB model as a functional predictive in vivo tool to select hit compounds, and suggest that enhancing BBB tightness and function has the potential to reduce Aβ pathology associated with AD.
登录
查看更多内容
三个紧密连接相关的Maguks ZO-1，ZO-2和ZO-3与Claudins的Cooh Termini直接结合。
DOI：
10.1083/jcb.147.6.1351
发表时间：
1999-12-13
期刊：
The Journal of cell biology
影响因子：
--
作者：
Itoh M;Furuse M;Morita K;Kubota K;Saitou M;Tsukita S
通讯作者：
Tsukita S
影响因子：
5
作者：
Qosa H;Batarseh YS;Mohyeldin MM;El Sayed KA;Keller JN;Kaddoumi A
通讯作者：
Kaddoumi A
影响因子：
5
作者：
Batarseh YS;Bharate SS;Kumar V;Kumar A;Vishwakarma RA;Bharate SB;Kaddoumi A
通讯作者：
Kaddoumi A
影响因子：
5
作者：
Abuznait, Alaa H.;Qosa, Hisham;Kaddoumi, Amal
通讯作者：
Kaddoumi, Amal
影响因子：
2.1
作者：
Galbusera, Carmen;Facheris, Maurizio;Ferrarese, Carlo
通讯作者：
Ferrarese, Carlo