功能性CRISPR分析揭示調控人調節性T細胞特性的基因網絡
作者：
小柯機器人發布時間：2020/9/30 15:10:50
美國加州大學Alexander Marson和德國慕尼黑工業大學Kathrin Schumann研究團隊合作取得一項新突破。他們利用功能性CRISPR篩選揭示了調控人調節性T細胞（Treg）身份的基因網絡。 2020年9月28日的《自然-免疫學》發表了這項成果。
在本研究中，研究人員使用合併和排列的Cas9核糖核蛋白篩選來鑑別在穩態和促炎條件下調控原代Treg細胞關鍵蛋白表達的轉錄因子。然後，研究人員在經歷遺傳擾動和細胞因子刺激的Treg細胞中，分析了54,424單細胞轉錄組。研究發現這些細胞除了受FOXO1和IRF4調控的網絡外，還分別受FOXP3和PRDM1單獨調控的基因網絡。之前認為HIVEP2與Treg細胞功能無關，它與SATB1共同調控了另一個基因網絡，對於Treg細胞介導的免疫抑制非常重要。
通過整合CRISPR篩選和單細胞RNA序列分析，研究人員揭示了人Treg細胞的轉錄調節因子和下遊基因網絡，這些網絡可作為免疫療法的潛在靶點。
據悉，Treg細胞對於免疫穩態至關重要。轉錄因子FOXP3維持Treg細胞的身份，但調控Treg細胞基因表達的關鍵轉錄因子合集仍然未知。
附：英文原文
Title: Functional CRISPR dissection of gene networks controlling human regulatory T cell identity
Author: Kathrin Schumann, Siddharth S. Raju, Michael Lauber, Saskia Kolb, Eric Shifrut, Jessica T. Cortez, Nikolaos Skartsis, Vinh Q. Nguyen, Jonathan M. Woo, Theodore L. Roth, Ruby Yu, Michelle L. T. Nguyen, Dimitre R. Simeonov, David N. Nguyen, Sasha Targ, Rachel E. Gate, Qizhi Tang, Jeffrey A. Bluestone, Matthew H. Spitzer, Chun Jimmie Ye, Alexander Marson
Issue&Volume: 2020-09-28
Abstract: Human regulatory T (Treg) cells are essential for immune homeostasis. The transcription factor FOXP3 maintains Treg cell identity, yet the complete set of key transcription factors that control Treg cell gene expression remains unknown. Here, we used pooled and arrayed Cas9 ribonucleoprotein screens to identify transcription factors that regulate critical proteins in primary human Treg cells under basal and proinflammatory conditions. We then generated 54,424 single-cell transcriptomes from Treg cells subjected to genetic perturbations and cytokine stimulation, which revealed distinct gene networks individually regulated by FOXP3 and PRDM1, in addition to a network coregulated by FOXO1 and IRF4. We also discovered that HIVEP2, to our knowledge not previously implicated in Treg cell function, coregulates another gene network with SATB1 and is important for Treg cell–mediated immunosuppression. By integrating CRISPR screens and single-cell RNA-sequencing profiling, we have uncovered transcriptional regulators and downstream gene networks in human Treg cells that could be targeted for immunotherapies. Treg cells are essential for immune homeostasis, but the transcription factors controlling their cellular identity are incompletely understood. Schumann and colleagues use pooled and arrayed CRISPR screens and scRNA-seq to describe key gene networks in human Treg cells.
DOI: 10.1038/s41590-020-0784-4
Source: https://www.nature.com/articles/s41590-020-0784-4