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Updated: Nov 19, 2025
Engineering Oncogenic Heterozygous Gain-of-Function Mutations in Human Hematopoietic Stem and Progenitor Cells
Published on: March 10, 2023
Dissecting ELANE neutropenia pathogenicity by human HSC gene editing
Shuquan Rao1, Yao Yao2, Josias Soares de Brito3
1Division of Hematology/Oncology, Boston Children's Hospital, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Stem Cell Institute, Broad Institute, Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA.
Gene editing can overcome neutrophil maturation defects in severe congenital neutropenia (SCN) caused by ELANE mutations. This approach restores normal neutrophil production and preserves hematopoietic stem and progenitor cell function.
Area of Science:
- Hematology
- Molecular Biology
- Genetics
Background:
- Severe congenital neutropenia (SCN) is a serious blood disorder.
- It is often caused by mutations in the ELANE gene, which disrupt neutrophil development.
- Current treatments aim to manage symptoms but not address the root cause.
Purpose of the Study:
- To investigate the use of CRISPR gene editing to correct ELANE mutations.
- To understand how different ELANE mutations affect neutrophil maturation.
- To develop a potential therapeutic strategy for ELANE-mutant SCN.
Main Methods:
- Conducted pooled CRISPR screening in human hematopoietic stem and progenitor cells (HSPCs).
- Correlated ELANE mutations with neutrophil maturation potential using gene editing.
- Established an animal model to study ELANE-mutant SCN.
Main Results:
- Early exon editing induced nonsense-mediated decay (NMD), restoring neutrophil maturation in patient HSPCs.
- Terminal exon frameshift alleles mimicked SCN mutations, causing maturation arrest.
- Identified specific frameshift types (-1 vs. -2 indels) that differentially impact neutrophil maturation.
Conclusions:
- Gene editing of ELANE mutations can restore normal neutrophil production.
- This approach preserves HSPC function and offers a potential universal therapy for ELANE-mutant neutropenia.
- CRISPR editing in HSPCs accurately identifies variant pathogenicity and clarifies disease mechanisms.
