Massively parallel base editing to map variant effects in human hematopoiesis

Area of Science:

• Genetics and Genomics
• Hematology
• Molecular Biology

Background:

• Systematic evaluation of genetic variants is crucial for understanding human health and disease.
• Current methods for introducing specific mutations are limited in primary cells like blood and immune cells.
• Scalable approaches for variant analysis in these critical cell types are needed.

Purpose of the Study:

• To develop and demonstrate massively parallel base-editing screens in human hematopoietic stem and progenitor cells.
• To enable functional screening of genetic variants across all hematopoietic differentiation states.
• To advance high-throughput variant-to-function mapping in human hematopoiesis.

Main Methods:

• Development of massively parallel base-editing screens.
• Application in human hematopoietic stem and progenitor cells.
• Phenotyping using single-cell RNA sequencing and editing outcome characterization via pooled single-cell genotyping.

Main Results:

• Efficient design of improved leukemia immunotherapy approaches.
• Comprehensive identification of non-coding variants affecting fetal hemoglobin expression.
• Definition of mechanisms regulating hematopoietic differentiation.
• Probing pathogenicity of uncharacterized disease-associated variants.

Conclusions:

• The developed base-editing screens provide a powerful tool for functional variant analysis in human hematopoiesis.
• These strategies facilitate the identification of genetic causes for diverse diseases.
• This approach advances the understanding and treatment of blood-related disorders.