Ablation of cDC2 development by triple mutations within the Zeb2 enhancer.
Ablation of cDC2 development by triple mutations within the Zeb2 enhancer.
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通过Zeb2增强子内的三重突变消除cDC2发育。
DOI：
10.1038/s41586-022-04866-z
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发表时间：
2022-07
期刊：
影响因子：
64.8
通讯作者：
中科院分区：
文献类型：
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作者：
常见的树突状细胞祖细胞（CDP）分化为传统的1型和2型树突状细胞（分别为cDC 1和cDC 2）谱系知之甚少。一些转录因子在已经指定的祖细胞的定型中起作用，例如BATF 3，其在+32 kb Irf 8增强子处稳定Irf 8自激活，但是控制CDP初始分化的机制仍然未知。在这里，我们报告的转录基础CDP分歧，并描述了前cDC 2规范的第一个要求。遗传上位性分析表明，Nfil 3在cDC 1发育中作用于Id 2、Batf 3和Zeb 2的上游，但没有揭示其机制或靶点。对新产生的NFIL 3报告小鼠的分析显示，在cDC 1特化期间NFIL 3表达极其短暂。CUT&RUN和染色质免疫沉淀，然后测序，鉴定了内源性NFIL 3在−165 kb Zeb 2增强子的三个位点结合，这些位点也结合CCAAT增强子结合蛋白C/EBPα和C/EBPβ。使用CRISPR-Cas9靶向的体内突变分析显示，这些NFIL 3-C/EBP位点在功能上是冗余的，C/EBP支持这些位点处的Zeb 2表达，而NFIL 3抑制这些位点处的Zeb 2表达。所有三个NFIL 3-C/EBP位点的三重突变消除了髓系祖细胞中的Zeb 2表达，但没有消除淋巴祖细胞中的Zeb 2表达，导致体内前cDC 2特化和成熟cDC 2发育的完全丧失。这些小鼠不产生针对多脑Heligmosomoides polygyrus感染的T辅助2（TH 2）细胞应答，这与cDC 2支持针对蠕虫的TH 2应答一致。因此，CDP分化为cDC 1或cDC 2是由NFIL 3和C/EBP在-165 kb Zeb 2增强子处的竞争控制的。
The divergence of the common dendritic cell progenitor (CDP) into the conventional type 1 and type 2 dendritic cell (cDC1 and cDC2, respectively) lineages is poorly understood. Some transcription factors act in the commitment of already specified progenitors–such as BATF3, which stabilizes Irf8 autoactivation at the +32 kb Irf8 enhancer–but the mechanisms controlling the initial divergence of CDPs remain unknown. Here we report the transcriptional basis of CDP divergence and describe the first requirements for pre-cDC2 specification. Genetic epistasis analysis suggested that Nfil3 acts upstream of Id2, Batf3 and Zeb2 in cDC1 development but did not reveal its mechanism or targets. Analysis of newly generated NFIL3 reporter mice showed extremely transient NFIL3 expression during cDC1 specification. CUT&RUN and chromatin immunoprecipitation followed by sequencing identified endogenous NFIL3 binding in the −165 kb Zeb2 enhancer at three sites that also bind the CCAAT-enhancer-binding proteins C/EBPα and C/EBPβ. In vivo mutational analysis using CRISPR-Cas9 targeting showed that these NFIL3–C/EBP sites are functionally redundant, with C/EBPs supporting and NFIL3 repressing Zeb2 expression at these sites. A triple mutation of all three NFIL3-C/EBP sites ablated Zeb2 expression in myeloid, but not lymphoid progenitors, causing the complete loss of pre-cDC2 specification and mature cDC2 development in vivo. These mice did not generate T helper 2 (TH2) cell responses against Heligmosomoides polygyrus infection, consistent with cDC2 supporting TH2 responses to helminths. Thus, CDP divergence into cDC1 or cDC2 is controlled by competition between NFIL3 and C/EBPs at the −165 kb Zeb2 enhancer.
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DOI：
10.1093/bioinformatics/btr064
发表时间：
2011-04-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Grant CE;Bailey TL;Noble WS
通讯作者：
Noble WS
NFIL3/E4BP4是在体内开发和成熟所必需的。
DOI：
10.1084/jem.20092176
发表时间：
2009-12-21
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Kamizono S;Duncan GS;Seidel MG;Morimoto A;Hamada K;Grosveld G;Akashi K;Lind EF;Haight JP;Ohashi PS;Look AT;Mak TW
通讯作者：
Mak TW
影响因子：
30.5
作者：
Grajales-Reyes GE;Iwata A;Albring J;Wu X;Tussiwand R;Kc W;Kretzer NM;Briseño CG;Durai V;Bagadia P;Haldar M;Schönheit J;Rosenbauer F;Murphy TL;Murphy KM
通讯作者：
Murphy KM
DOI：
10.4049/jimmunol.1102613
发表时间：
2012-10-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Bajaña S;Roach K;Turner S;Paul J;Kovats S
通讯作者：
Kovats S
影响因子：
30.5
作者：
Iwata A;Durai V;Tussiwand R;Briseño CG;Wu X;Grajales-Reyes GE;Egawa T;Murphy TL;Murphy KM
通讯作者：
Murphy KM