小柯机器人

美国加州大学Martin Kampmann和美国格莱斯顿神经疾病研究所Li Gan共同合作，近期取得重要工作进展。他们研究利用人类iPSC来源小胶质细胞CRISPRi/a筛选平台揭示了疾病状态的调节因子。相关论文2022年8月11日在线发表于《自然—神经科学》杂志上。

研究人员提出了一个筛选平台，以系统地阐明遗传扰动在人类诱导的多能干细胞来源的小胶质细胞中的功能影响。他们基于六种转录因子的诱导表达开发了一种高效的8天生成小胶质细胞样细胞的方案。研究人员在该系统中建立了诱导型CRISPR干扰和激活，并针对“可药用基因组”进行了三个筛选。

这些筛选揭示了控制小胶质细胞存活、激活和吞噬作用的基因，包括神经退行性变相关基因。以单细胞RNA测序作为读数的筛选显示，这些小胶质细胞采用了一系列与人类大脑中观察到的状态相似的状态，并确定了这些状态的调节因子。以骨桥蛋白（SPP1）表达为特征的疾病相关状态被集落刺激因子-1（CSF1R）抑制选择性地耗尽。因此，他们的平台可以系统地揭示小胶质细胞状态的调控因子，使其功能表征和治疗靶向成为可能。

据介绍，小胶质细胞正在成为神经系统疾病的关键驱动因素，然而对于潜在的机制目前还缺乏系统的理解。

附：英文原文

Title: A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states

Author: Drger, Nina M., Sattler, Sydney M., Huang, Cindy Tzu-Ling, Teter, Olivia M., Leng, Kun, Hashemi, Sayed Hadi, Hong, Jason, Aviles, Giovanni, Clelland, Claire D., Zhan, Lihong, Udeochu, Joe C., Kodama, Lay, Singleton, Andrew B., Nalls, Mike A., Ichida, Justin, Ward, Michael E., Faghri, Faraz, Gan, Li, Kampmann, Martin

Issue&Volume: 2022-08-11

Abstract: Microglia are emerging as key drivers of neurological diseases. However, we lack a systematic understanding of the underlying mechanisms. Here, we present a screening platform to systematically elucidate functional consequences of genetic perturbations in human induced pluripotent stem cell-derived microglia. We developed an efficient 8-day protocol for the generation of microglia-like cells based on the inducible expression of six transcription factors. We established inducible CRISPR interference and activation in this system and conducted three screens targeting the ‘druggable genome’. These screens uncovered genes controlling microglia survival, activation and phagocytosis, including neurodegeneration-associated genes. A screen with single-cell RNA sequencing as the readout revealed that these microglia adopt a spectrum of states mirroring those observed in human brains and identified regulators of these states. A disease-associated state characterized by osteopontin (SPP1) expression was selectively depleted by colony-stimulating factor-1 (CSF1R) inhibition. Thus, our platform can systematically uncover regulators of microglial states, enabling their functional characterization and therapeutic targeting.

DOI: 10.1038/s41593-022-01131-4

Source: https://www.nature.com/articles/s41593-022-01131-4

Nature Neuroscience：《自然—神经科学》，创刊于1998年。隶属于施普林格·自然出版集团，最新IF：28.771
官方网址：https://www.nature.com/neuro/
投稿链接：https://mts-nn.nature.com/cgi-bin/main.plex