In vivo CRISPR screening identifies BAZ2 chromatin remodelers as druggable regulators of mammalian liver regeneration.
In vivo CRISPR screening identifies BAZ2 chromatin remodelers as druggable regulators of mammalian liver regeneration.
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DOI：
10.1016/j.stem.2022.01.001
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发表时间：
2022-03-03
期刊：
影响因子：
23.9
通讯作者：
Zhu H
中科院分区：
文献类型：
--
作者：
Jia Y;Li L;Lin YH;Gopal P;Shen S;Zhou K;Yu X;Sharma T;Zhang Y;Siegwart DJ;Ready JM;Zhu H
由于缺乏体内筛选方法，确定调节哺乳动物再生的新途径具有挑战性。我们在再生肝脏中采用合并的CRISPR敲除和活化筛选来评估165种染色质调节蛋白。这两个筛选鉴定了模仿-SWI染色质重塑组分Baz 2a和Baz 2b，其先前未涉及再生。体内sgRNA，siRNA，和敲除策略，对任一paranesthesia证实增加再生。不同的BAZ 2特异性布罗莫结构域抑制剂GSK 2801和BAZ 2-ICR导致不同损伤后的加速肝脏愈合。抑制剂治疗的小鼠在炎症性肠病模型中也表现出改善的愈合，表明多组织适用性。再生肝脏的转录组学显示核糖体和细胞周期mRNA增加。令人惊讶的是，CRISPRa筛选以定义机制显示，过量产生RpllOa或Rpl 24足以驱动再生，而Rpl 24单倍不足是BAZ 2抑制介导的再生的速率限制。模拟SWI成分的再生作用的发现为增强组织修复提供了直接的策略。在哺乳动物模型中识别新的组织再生调节剂具有挑战性。在此，肝脏中的合并体内CRISPR筛选鉴定了模拟SWI染色质重塑组分Baz 2a和Baz 2b作为再生抑制剂。BAZ 2特异性布罗莫结构域抑制剂导致肝脏和肠道愈合加速，其作用通过再生过程中蛋白质合成的增加来介导。
Identifying new pathways that regulate mammalian regeneration is challenging due to the paucity of in vivo screening approaches. We employed pooled CRISPR knockout and activation screening in the regenerating liver to evaluate 165 chromatin regulatory proteins. Both screens identified imitation-SWI chromatin remodeling components Baz2a and Baz2b, not previously implicated in regeneration. In vivo sgRNA, siRNA, and knockout strategies against either paralog confirmed increased regeneration. Distinct BAZ2-specific bromodomain inhibitors GSK2801 and BAZ2-ICR resulted in accelerated liver healing after diverse injuries. Inhibitor treated mice also exhibited improved healing in an inflammatory bowel disease model, suggesting multi-tissue applicability. Transcriptomics on regenerating livers showed increases in ribosomal and cell cycle mRNAs. Surprisingly, CRISPRa screening to define mechanisms showed that overproducing Rpl10a or Rpl24 was sufficient to drive regeneration, while Rpl24 haploinsufficiency was rate limiting for BAZ2 inhibition mediated regeneration. The discovery of regenerative roles for imitation-SWI components provides immediate strategies to enhance tissue repair. New regulators of tissue regeneration are challenging to identify in mammalian models. Here, pooled in vivo CRISPR screening in the liver identified imitation-SWI chromatin remodeling components Baz2a and Baz2b as regeneration suppressors. BAZ2-specific bromodomain inhibitors resulted in accelerated liver and intestinal healing, effects mediated through increased protein synthesis during regeneration.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
DOI：
10.15252/embj.2020105606
发表时间：
2020-12-01
期刊：
The EMBO journal
影响因子：
--
作者：
Dalcher D;Tan JY;Bersaglieri C;Peña-Hernández R;Vollenweider E;Zeyen S;Schmid MW;Bianchi V;Butz S;Roganowicz M;Kuzyakiv R;Baubec T;Marques AC;Santoro R
通讯作者：
Santoro R
影响因子：
30.8
作者：
GROMPE, M;LINDSTEDT, S;FINEGOLD, M
通讯作者：
FINEGOLD, M
影响因子：
7.2
作者：
Liang, Roger;Lin, Yu-Hsuan;Zhu, Hao
通讯作者：
Zhu, Hao
影响因子：
64.8
作者：
Blanco, Sandra;Bandiera, Roberto;Popis, Martyna;Hussain, Shobbir;Lombard, Patrick;Aleksic, Jelena;Sajini, Abdulrahim;Tanna, Hinal;Cortes-Garrido, Rosana;Gkatza, Nikoletta;Dietmann, Sabine;Frye, Michaela
通讯作者：
Frye, Michaela