FOCAL ADHESION KINASE ACTIVATION IN THE BLOOD-BRAIN BARRIER

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在HIV脑炎（HIVE）的发展过程中，血脑屏障（BBB）的脑微血管内皮细胞（MBEC）之间的紧密连接蛋白的表达丢失。 虽然许多研究集中在诱导神经系统后遗症的病毒株或与脑炎发展相关的巨噬细胞/小胶质细胞上，但涉及的BMEC内的分子信号传导途径尚未得到解决。 我们以前已经表明，有激活和破坏的体外血脑屏障模型使用慢病毒感染的CEMx 174细胞。 我们和其他人已经在体内显示了类似的破坏。 因此，感兴趣的是确定感染的原代巨噬细胞的存在是否可以立即离体破坏完整的脑微血管，如果是这样，则涉及哪些信号通路。 我们的数据表明，BMEC之间的紧密连接的破坏是通过在TYR-397磷酸化激活粘着斑激酶（FAK）介导的。 我们还表明，肌球蛋白轻链激酶（MLCK）也磷酸化在同一时间过程中作为FAK。 抑制FAK的活化足以防止紧密连接的破坏。因此，有可能通过使用FAK和MLCK的抑制剂来抑制HIVE的发展。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Expression of tight junction proteins between brain microvascular endothelial cells (MBEC) of the blood-brain barrier (BBB) is lost during development of HIV encephalitis (HIVE). While many studies have focused on the strains of virus that induce neurological sequelae or on the macrophages/microglia that are associated with the development of encephalitis, the molecular signaling pathways within the BMECs involved have yet to be resolved. We have previously shown that there is activation and disruption of an in vitro BBB model using lentivirus-infected CEMx174 cells. We and others have shown similar disruption in vivo. Therefore, it was of interest to determine of the presence of infected primary macrophages could disrupt intact cerebral microvessels immediately ex vivo, and if so, which signaling pathways were involved. Our data demonstrate that disruption of tight junctions between BMECs is mediated through activation of focal adhesion kinase (FAK) by phosphorylation at TYR-397. We also show that myosin light chain kinase (MLCK) is also phosphorylated in the same time course as FAK. Inhibition of FAK activation is sifficient to prevent tight junction disruption. This, it may be possible to inhibit the development of HIVE by using inhibitors of FAK and MLCK.