Engineering monocyte/macrophage-specific glucocerebrosidase expression in human hematopoietic stem cells using genome...

Area of Science:

• Genetics and Gene Therapy
• Hematology
• Lysosomal Storage Disorders

Background:

• Gaucher disease results from deficient glucocerebrosidase activity, leading to macrophage accumulation and inflammation.
• Current treatments involve enzyme replacement or enzyme inhibitors, requiring lifelong administration.
• Cellular engineering offers a potential one-time curative approach for Gaucher disease.

Purpose of the Study:

• To develop an efficient CRISPR/Cas9-based gene editing strategy for Gaucher disease.
• To target glucocerebrosidase expression to monocytes/macrophages using a safe-harbor locus.
• To evaluate the potential for a one-time, curative therapy for Gaucher disease.

Main Methods:

• Utilized CRISPR/Cas9 gene editing to insert glucocerebrosidase expression cassettes into the CCR5 safe-harbor locus.
• Targeted human hematopoietic stem and progenitor cells (HSPCs).
• Assessed long-term repopulation and multi-lineage differentiation potential through serial transplantation.

Main Results:

• Successfully generated macrophages expressing functional glucocerebrosidase from edited HSPCs.
• Demonstrated sustained repopulation and multi-lineage differentiation capacity in transplanted cells.
• Validated the CCR5 locus as a suitable safe-harbor for lineage-specific gene expression.

Conclusions:

• CRISPR/Cas9-mediated gene insertion into the CCR5 locus provides a universal strategy for Gaucher disease correction.
• This approach enables macrophage-specific glucocerebrosidase expression, avoiding stem cell toxicity.
• The platform is adaptable for treating other lysosomal storage disorders.