Internal medicine

In vivo

Biology

Gene

Haematopoiesis

Genetic enhancement

Globin

Paleontology

Immunology

Ex vivo

Anemia

Stem cell

Myeloid

Context (archaeology)

Erythropoiesis

Cell biology

CRISPR

Transplantation

Cancer research

Genetics

Medicine

Nature Communications

CRISPR/Cas9-mediated beta-globin ( HBB ) gene correction of sickle cell disease (SCD) patient-derived hematopoietic stem cells (HSCs) in combination with autologous transplantation represents a recent paradigm in gene therapy. Although several Cas9-based HBB -correction approaches have been proposed, functional correction of in vivo erythropoiesis has not been investigated previously. Here, we use a humanized globin-cluster SCD mouse model to study Cas9-AAV6-mediated HBB- correction in functional HSCs within the context of autologous transplantation. We discover that long-term multipotent HSCs can be gene corrected ex vivo and stable hemoglobin-A production can be achieved in vivo from HBB -corrected HSCs following autologous transplantation. We observe a direct correlation between increased HBB -corrected myeloid chimerism and normalized in vivo red blood cell (RBC) features, but even low levels of chimerism resulted in robust hemoglobin-A levels. Moreover, this study offers a platform for gene editing of mouse HSCs for both basic and translational research.

Cas9-AAV6 gene correction of beta-globin in autologous HSCs improves sickle cell disease erythropoiesis in mice