A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states.
A CRISPRi/a platform in human iPSC-derived microglia uncovers regulators of disease states.
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DOI：
10.1038/s41593-022-01131-4
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发表时间：
2022-09
影响因子：
25
通讯作者：
Kampmann, Martin
中科院分区：
文献类型：
--
作者：
Drager, 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
小胶质细胞正在成为神经系统疾病的关键驱动因素。然而，我们缺乏对潜在机制的系统理解。在这里，我们提出了一个筛选平台，以系统地阐明遗传扰动在人类诱导多能干细胞衍生的小胶质细胞中的功能后果。我们基于六种转录因子的诱导表达，开发了一种高效的8天小胶质样细胞生成方案。我们在该系统中建立了可诱导的CRISPR干扰和激活，并针对“可药物基因组”进行了三次筛选。这些筛选揭示了控制小胶质细胞存活、激活和吞噬的基因，包括神经变性相关基因。一个单细胞RNA测序的屏幕显示，这些小胶质细胞采用了与人类大脑中观察到的状态光谱，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，我们的平台可以系统地揭示小胶质细胞状态的调节因子，使其功能表征和治疗靶向成为可能。Dräger等人建立了一个快速、可扩展的ipsc衍生小胶质细胞平台。CRISPRi/a筛选揭示了疾病相关基因在吞噬作用中的作用，以及疾病相关小胶质细胞状态的调节因子，这些可以在药理学上靶向。
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. Dräger et al. establish a rapid, scalable platform for iPSC-derived microglia. CRISPRi/a screens uncover roles of disease-associated genes in phagocytosis, and regulators of disease-relevant microglial states that can be targeted pharmacologically.
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影响因子：
5.9
作者：
Chen SW;Hung YS;Fuh JL;Chen NJ;Chu YS;Chen SC;Fann MJ;Wong YH
通讯作者：
Wong YH
影响因子：
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作者：
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通讯作者：
Schlesner, Matthias
影响因子：
5.9
作者：
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通讯作者：
Fossati V
影响因子：
32.4
作者：
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通讯作者：
Stevens, Beth
影响因子：
12.7
作者：
Bliederhaeuser, Corinna;Grozdanov, Veselin;Danzer, Karin M.
通讯作者：
Danzer, Karin M.