Seo Joon Pyung, Trippett James S, Huang Zhimin, Wang Ryan Z, Lee Sangsin, Szablowski Jerzy O
bioRxiv. 2023 May 24:2023.05.23.541868. doi: 10.1101/2023.05.23.541868.
Gene expression is a critical component of brain physiology and activity, but monitoring this expression in the living brain represents a significant challenge. Here, we introduce a new paradigm called Recovery of Markers through InSonation (REMIS) for noninvasive measurement of gene expression in the brain with cell-type, spatial, and temporal specificity. Our approach relies on engineered protein markers that are designed to be expressed in neurons and exit into the interstitium. By applying ultrasound to targeted brain regions, these markers are released into the bloodstream, where they can be readily detected using biochemical techniques. REMIS can noninvasively confirm gene delivery and measure endogenous signaling in specific brain sites through a simple insonation and a subsequent blood test. Using REMIS, we successfully measured chemogenetic induction of neuronal activity in ultrasound-tar-geted brain regions. REMIS recovery of markers is reliable and demonstrated improved recovery of markers from the brain into the blood in every tested animal. Overall, our work establishes a noninvasive, spatially-specific means of monitoring gene delivery outcomes and endogenous signaling in mammalian brains, opening up possibilities for brain research and noninvasive monitoring of gene therapies in the brain.
基因表达是大脑生理和活动的关键组成部分，但在活体大脑中监测这种表达是一项重大挑战。在此，我们引入了一种名为“通过超声检测恢复标记物”（REMIS）的新范式，用于对大脑中的基因表达进行具有细胞类型、空间和时间特异性的非侵入性测量。我们的方法依赖于经过工程改造的蛋白质标记物，这些标记物被设计用于在神经元中表达并释放到间质中。通过将超声应用于目标脑区，这些标记物被释放到血液中，在那里可以使用生化技术轻松检测到它们。REMIS可以通过简单的超声检测和随后的血液检测，非侵入性地确认基因传递并测量特定脑区的内源性信号传导。使用REMIS，我们成功地测量了超声靶向脑区中神经元活动的化学遗传诱导。REMIS标记物的恢复是可靠的，并且在每只受试动物中都显示出从大脑到血液中标记物的恢复有所改善。总体而言，我们的工作建立了一种非侵入性的、空间特异性的方法来监测哺乳动物大脑中的基因传递结果和内源性信号传导，为大脑研究和大脑基因治疗的非侵入性监测开辟了可能性。
