小柯机器人

切换到桌面版
Zeb2增强子内三重突变对cDC2发育的抑制作用
2022-06-26 15:16
来源：小柯机器人

美国圣路易斯华盛顿大学Kenneth M. Murphy研究组发现，Zeb2增强子内的三重突变削弱cDC2的发育。该研究于2022年6月22日在线发表于国际一流学术期刊《自然》。

研究人员表示，普通树突状细胞前体（CDP）分化为传统的1型和2型树突状细胞（分别为cDC1和cDC2）谱系的过程还不太清楚。一些转录因子在已经特定前体细胞的确立中起作用，如BATF3，它在+32 kb的Irf8增强子上稳定Irf8的自动激活，但控制CDP最初分化的机制仍然未知。
 
研究人员报告了CDP分化的转录基础，并描述了cDC2前特化的第一个要求。遗传上位法分析表明，Nfil3在cDC1的发育中处于Id2、Batf3和Zeb2的上游，但没有揭示其机制或靶标。对新产生的NFIL3报告小鼠的分析表明，NFIL3在cDC1发育过程中表达极为短暂。CUT&RUN和染色质免疫沉淀后的测序确定了NFIL3在-165kb的Zeb2增强子中的内源性结合，这三个位点也与CCAAT-增强子结合蛋白C/EBPα和C/EBPβ结合。
 
使用CRISPR-Cas9靶向的体内突变分析表明，这些NFIL3-C/EBP位点在功能上是多余的，C/EBP支持而NFIL3抑制Zeb2在这些位点的表达。所有三个NFIL3-C/EBP位点的三重突变使Zeb2在骨髓，而不是淋巴祖细胞中的表达消失，导致体内pre-cDC2特化和成熟cDC2发育的完全丧失。这些小鼠对蠕虫Heligmosomoides polygyrus感染没有产生T辅助2（TH2）细胞反应，这与cDC2支持TH2对蠕虫的反应相一致。因此，CDP分化为cDC1或cDC2是由NFIL3和C/EBPs在-165 kb Zeb2增强子上的竞争所控制。
 
附：英文原文
 
Title: Ablation of cDC2 development by triple mutations within the Zeb2 enhancer

Author: Liu, Tian-Tian, Kim, Sunkyung, Desai, Pritesh, Kim, Do-Hyun, Huang, Xiao, Ferris, Stephen T., Wu, Renee, Ou, Feiya, Egawa, Takeshi, Van Dyken, Steven J., Diamond, Michael S., Johnson, Peter F., Kubo, Masato, Murphy, Theresa L., Murphy, Kenneth M.

Issue&Volume: 2022-06-22

Abstract: The divergence of the common dendritic cell progenitor1,2,3 (CDP) into the conventional type 1 and type 2 dendritic cell (cDC1 and cDC2, respectively) lineages4,5 is poorly understood. Some transcription factors act in the commitment of already specified progenitors—such as BATF3, which stabilizes Irf8 autoactivation at the +32kb Irf8 enhancer4,6—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 analysis7 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 –165kb Zeb2 enhancer8 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 helminths9,10,11. Thus, CDP divergence into cDC1 or cDC2 is controlled by competition between NFIL3 and C/EBPs at the –165kb Zeb2 enhancer.

DOI: 10.1038/s41586-022-04866-z

Source: https://www.nature.com/articles/s41586-022-04866-z

Nature：《自然》，创刊于1869年。隶属于施普林格·自然出版集团，最新IF：69.504
官方网址：http://www.nature.com/
投稿链接：http://www.nature.com/authors/submit_manuscript.html


本期文章：《自然》：Online/在线发表

分享到:

0