- Shi, Jianting;
- Wu, Xun;
- Wang, Ziyi;
- Li, Fang;
- Meng, Yujiao;
- Moore, Rebecca M;
- Cui, Jian;
- Xue, Chenyi;
- Croce, Katherine R;
- Yurdagul, Arif;
- Doench, John G;
- Li, Wei;
- Zarbalis, Konstantinos S;
- Tabas, Ira;
- Yamamoto, Ai;
- Zhang, Hanrui
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.