Pooled CRISPR screening identifies m(6)A as a positive regulator of macrophage activation.
Pooled CRISPR screening identifies m(6)A as a positive regulator of macrophage activation.
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汇集 CRISPR 筛选鉴定 m 6 A 为巨噬细胞激活的正调节因子
DOI：
10.1126/sciadv.abd4742
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发表时间：
2021-04
期刊：
影响因子：
13.6
通讯作者：
Li HB
中科院分区：
文献类型：
--
作者：
Tong J;Wang X;Liu Y;Ren X;Wang A;Chen Z;Yao J;Mao K;Liu T;Meng FL;Pan W;Zou Q;Liu J;Zhou Y;Xia Q;Flavell RA;Zhu S;Li HB
CRISPR筛选鉴定RNA m6 A修饰为TLR信号通路的负调节剂，用于适当的先天免疫应答。m6 A RNA修饰涉及多种细胞反应。然而，其在先天免疫细胞中的功能知之甚少。在这里，我们在RNA结合蛋白聚焦CRISPR筛选中将主要的m6 A“作者”确定为调节LPS引起的巨噬细胞活化的最佳候选基因。我们已经证实，Mettl 3缺陷型巨噬细胞在体外LPS刺激后表现出TNF-α产生减少。一致地，Mettl 3flox/flox;Lyzm-Cre小鼠显示出对细菌感染的易感性增加，并且显示出更快的肿瘤生长。从机制上讲，编码TLR 4信号负调节因子的Irakm基因的转录本受到m6 A修饰的高度修饰。胃L3缺乏导致m6 A修饰的缺失，并减缓其降解，导致更高水平的IRAKM，最终抑制TLR信号介导的巨噬细胞活化。我们的研究结果表明，以前未知的作用，胃L3介导的m6 A修饰的先天免疫反应，并牵连m6 A机制作为一个潜在的癌症免疫治疗的目标。
CRISPR screening identifies RNA m6A modification as negative regulator of TLR signaling pathway for proper innate immune response. m6A RNA modification is implicated in multiple cellular responses. However, its function in the innate immune cells is poorly understood. Here, we identified major m6A “writers” as the top candidate genes regulating macrophage activation by LPS in an RNA binding protein focused CRISPR screening. We have confirmed that Mettl3-deficient macrophages exhibited reduced TNF-α production upon LPS stimulation in vitro. Consistently, Mettl3flox/flox;Lyzm-Cre mice displayed increased susceptibility to bacterial infection and showed faster tumor growth. Mechanistically, the transcripts of the Irakm gene encoding a negative regulator of TLR4 signaling were highly decorated by m6A modification. METTL3 deficiency led to the loss of m6A modification on Irakm mRNA and slowed down its degradation, resulting in a higher level of IRAKM, which ultimately suppressed TLR signaling–mediated macrophage activation. Our findings demonstrate a previously unknown role for METTL3-mediated m6A modification in innate immune responses and implicate the m6A machinery as a potential cancer immunotherapy target.
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影响因子：
10.5
作者：
Lasman L;Krupalnik V;Viukov S;Mor N;Aguilera-Castrejon A;Schneir D;Bayerl J;Mizrahi O;Peles S;Tawil S;Sathe S;Nachshon A;Shani T;Zerbib M;Kilimnik I;Aigner S;Shankar A;Mueller JR;Schwartz S;Stern-Ginossar N;Yeo GW;Geula S;Novershtern N;Hanna JH
通讯作者：
Hanna JH
DOI：
10.1038/nrclinonc.2016.217
发表时间：
2017-07
期刊：
Nature reviews. Clinical oncology
影响因子：
--
作者：
Mantovani A;Marchesi F;Malesci A;Laghi L;Allavena P
通讯作者：
Allavena P
影响因子：
32.4
作者：
Noy, Roy;Pollard, Jeffrey W.
通讯作者：
Pollard, Jeffrey W.
影响因子：
56.9
作者：
Liu, Jun;Do, Xiaoyang;Hei, Chuan
通讯作者：
Hei, Chuan
DOI：
10.4049/jimmunol.1402377
发表时间：
2015-02-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Ballinger MN;Newstead MW;Zeng X;Bhan U;Mo XM;Kunkel SL;Moore BB;Flavell R;Christman JW;Standiford TJ
通讯作者：
Standiford TJ