Laboratory of Molecular Neuro-Oncology and Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, 407 East 61(st) Street, New York, NY 10065, USA.
Cell Rep. 2019 Jul 23;28(4):979-991.e6. doi: 10.1016/j.celrep.2019.06.075.
Post-transcriptional regulation by microRNAs (miRNAs) is essential for complex molecular responses to physiological insult and disease. Although many disease-associated miRNAs are known, their global targets and culminating network effects on pathophysiology remain poorly understood. We applied Argonaute (AGO) crosslinking immunoprecipitation (CLIP) to systematically elucidate altered miRNA-target interactions in brain following ischemia and reperfusion (I/R) injury. Among 1,190 interactions identified, the most prominent was the cumulative loss of target regulation by miR-29 family members. Integration of translational and time-course RNA profiles revealed a dynamic mode of miR-29 target de-regulation, led by acute translational activation and a later increase in RNA levels, allowing rapid proteomic changes to take effect. These functional regulatory events rely on canonical and non-canonical miR-29 binding and engage glutamate reuptake signals, such as glial glutamate transporter (GLT-1), to control local glutamate levels. These results uncover a miRNA target network that acts acutely to maintain brain homeostasis after ischemic stroke.
miRNAs（微小 RNA）的转录后调控对于对生理损伤和疾病的复杂分子反应至关重要。尽管已知许多与疾病相关的 miRNAs，但它们对病理生理学的全局靶标和最终网络效应仍知之甚少。我们应用 Argonaute（AGO）交联免疫沉淀（CLIP）技术，系统地阐明了脑缺血再灌注（I/R）损伤后 miRNA 靶标相互作用的改变。在鉴定的 1190 种相互作用中，最突出的是 miR-29 家族成员的靶标调控的累积丧失。翻译和时间过程 RNA 谱的整合揭示了 miR-29 靶标去调控的动态模式，由急性翻译激活和随后 RNA 水平的增加引起，从而使快速的蛋白质组学变化生效。这些功能调节事件依赖于规范和非规范的 miR-29 结合，并涉及谷氨酸再摄取信号，如神经胶质谷氨酸转运蛋白（GLT-1），以控制局部谷氨酸水平。这些结果揭示了一个 miRNA 靶标网络，它在缺血性中风后急性作用以维持脑内稳态。
