| Autor: |
Rao, Shuquan, Yao, Yao, Soares de Brito, Josias, Yao, Qiuming, Shen, Anne H., Watkinson, Ruth E., Kennedy, Alyssa L., Coyne, Steven, Ren, Chunyan, Zeng, Jing, Serbin, Anna Victoria, Studer, Sabine, Ballotti, Kaitlyn, Harris, Chad E., Luk, Kevin, Stevens, Christian S., Armant, Myriam, Pinello, Luca, Wolfe, Scot A., Chiarle, Roberto, Shimamura, Akiko, Lee, Benhur, Newburger, Peter E., Bauer, Daniel E. |
| Zdroj: |
Cell Stem Cell; May 2021, Vol. 28 Issue: 5 p833-845.e5 |
| Abstrakt: |
Severe congenital neutropenia (SCN) is a life-threatening disorder most often caused by dominant mutations of ELANEthat interfere with neutrophil maturation. We conducted a pooled CRISPR screen in human hematopoietic stem and progenitor cells (HSPCs) that correlated ELANEmutations with neutrophil maturation potential. Highly efficient gene editing of early exons elicited nonsense-mediated decay (NMD), overcame neutrophil maturation arrest in HSPCs from ELANE-mutant SCN patients, and produced normal hematopoietic engraftment function. Conversely, terminal exon frameshift alleles that mimic SCN-associated mutations escaped NMD, recapitulated neutrophil maturation arrest, and established an animal model of ELANE-mutant SCN. Surprisingly, only −1 frame insertions or deletions (indels) impeded neutrophil maturation, whereas −2 frame late exon indels repressed translation and supported neutrophil maturation. Gene editing of primary HSPCs allowed faithful identification of variant pathogenicity to clarify molecular mechanisms of disease and encourage a universal therapeutic approach to ELANE-mutant neutropenia, returning normal neutrophil production and preserving HSPC function. |
| Databáze: |
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