A genome-wide CRISPR screen identifies WDFY3 as a regulator of macrophage efferocytosis.
A genome-wide CRISPR screen identifies WDFY3 as a regulator of macrophage efferocytosis.
复制标题
全基因组CRISPR筛查发现WDFY3是巨噬细胞泡泡吞噬的调节因子。
DOI：
10.1038/s41467-022-35604-8
复制
发表时间：
2022-12-24
影响因子：
16.6
通讯作者：
Zhang, Hanrui
中科院分区：
文献类型：
--
作者：
Shi, Jianting;Wu, Xun;Wang, Ziyi;Li, Fang;Meng, Yujiao;Moore, Rebecca M.;Cui, Jian;Xue, Chenyi;Croce, Katherine R.;Yurdagul, Arif, Jr.;Doench, John G.;Li, Wei;Zarbalis, Konstantinos S.;Tabas, Ira;Yamamoto, Ai;Zhang, Hanrui
吞噬细胞对垂死细胞的清除（称为胞吐作用）对于维持组织稳态至关重要，但我们对胞吐作用调节的理解仍然不完整。在这里，我们在原代小鼠巨噬细胞中进行了基于FACS的全基因组CRISPR敲除筛选，以寻找新的巨噬细胞调节剂。结果表明，巨噬细胞中的Wdfy 3敲除特异性地损害凋亡细胞的摄取，但不结合，这是由于缺陷的肌动蛋白分解。此外，WDFY 3与GABARAP相互作用，从而促进LC 3脂化和随后的溶酶体酸化，以允许凋亡细胞组分的降解。从机制上讲，虽然WDFY 3的C-末端足以挽救由Wdfy 3敲除诱导的受损降解，但需要全长WDFY 3来重建凋亡细胞的摄取。最后，在小鼠体内和体外在原代人巨噬细胞中，WDFY 3也是有效的巨噬细胞作用所必需的。因此，这项工作扩展了我们对巨噬细胞吞噬机制的了解，并支持全基因组CRISPR筛选作为询问初级巨噬细胞中复杂功能表型的平台。胞饮作用描述了吞噬细胞对凋亡细胞的吞噬和清除。在这里，作者在原代小鼠巨噬细胞中鉴定了WDFY 3作为细胞凋亡的调节剂，其中c-末端WDFY 3足以调节降解，而全长WDFY 3需要调节凋亡细胞的摄取。
Phagocytic clearance of dying cells, termed efferocytosis, is essential for maintaining tissue homeostasis, yet our understanding of efferocytosis regulation remains incomplete. Here we perform a FACS-based, genome-wide CRISPR knockout screen in primary mouse macrophages to search for novel regulators of efferocytosis. The results show that Wdfy3 knockout in macrophages specifically impairs uptake, but not binding, of apoptotic cells due to defective actin disassembly. Additionally, WDFY3 interacts with GABARAP, thus facilitating LC3 lipidation and subsequent lysosomal acidification to permit the degradation of apoptotic cell components. Mechanistically, while the C-terminus of WDFY3 is sufficient to rescue the impaired degradation induced by Wdfy3 knockout, full-length WDFY3 is required to reconstitute the uptake of apoptotic cells. Finally, WDFY3 is also required for efficient efferocytosis in vivo in mice and in vitro in primary human macrophages. This work thus expands our knowledge of the mechanisms of macrophage efferocytosis, as well as supports genome-wide CRISPR screen as a platform for interrogating complex functional phenotypes in primary macrophages. Efferocytosis describes the engulfment and clearance of apoptotic cells by phagocytes. Here the authors identify in primary mouse macrophage WDFY3 as a regulator for efferocytosis, in which c-terminal WDFY3 is sufficient to modulate degradation while full-length WDFY3 is required to modulate the uptake of apoptotic cells.
登录
查看更多内容
影响因子：
37.8
作者：
Kojima Y;Weissman IL;Leeper NJ
通讯作者：
Leeper NJ
影响因子：
12.4
作者：
Green DR;Oguin TH;Martinez J
通讯作者：
Martinez J
影响因子：
46.9
作者：
Doench JG;Fusi N;Sullender M;Hegde M;Vaimberg EW;Donovan KF;Smith I;Tothova Z;Wilen C;Orchard R;Virgin HW;Listgarten J;Root DE
通讯作者：
Root DE
影响因子：
30.8
作者：
Haney MS;Bohlen CJ;Morgens DW;Ousey JA;Barkal AA;Tsui CK;Ego BK;Levin R;Kamber RA;Collins H;Tucker A;Li A;Vorselen D;Labitigan L;Crane E;Boyle E;Jiang L;Chan J;Rincón E;Greenleaf WJ;Li B;Snyder MP;Weissman IL;Theriot JA;Collins SR;Barres BA;Bassik MC
通讯作者：
Bassik MC
影响因子：
16.2
作者：
Fox, Leora M.;Kim, Kiryung;Yamamoto, Ai
通讯作者：
Yamamoto, Ai