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

• Published in: Nature communications Vol. 12; no. 1; p. 686
• Main Authors: Wilkinson, Adam C., Dever, Daniel P., Baik, Ron, Camarena, Joab, Hsu, Ian, Charlesworth, Carsten T., Morita, Chika, Nakauchi, Hiromitsu, Porteus, Matthew H.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 29.01.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 13/100; 42; 42/41; 631/337/4041/3196; 631/532/1542; 631/61/201/2110; Anemia, Sickle Cell - blood; Anemia, Sickle Cell - diagnosis; Anemia, Sickle Cell - genetics; Anemia, Sickle Cell - therapy; Animals; Autografts; beta-Globins - genetics; Chimerism; Combined Modality Therapy - methods; Context; CRISPR; CRISPR-Cas Systems - genetics; Dependovirus; Disease Models, Animal; Erythrocytes; Erythropoiesis; Erythropoiesis - genetics; Female; Gene Editing - methods; Gene Knock-In Techniques; Gene therapy; Genetic modification; Genetic Therapy - methods; Genetic Vectors - genetics; Genome editing; Hematopoietic Stem Cell Transplantation - methods; Hematopoietic stem cells; Hemoglobin; Humanities and Social Sciences; Humans; In vivo methods and tests; Mice; Mice, Transgenic; multidisciplinary; Parvovirinae - genetics; Science; Science (multidisciplinary); Sickle cell disease; Stem cell transplantation; Stem cells; Transplantation; Transplantation, Autologous - methods
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-20909-x

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. CRISPR mediated gene correction of sickle cell disease (SCD) in patient-derived hematopoietic stem cells is a promising avenue for therapy. Here the authors use a humanized SCD mouse model to study gene editing in the context of autologous transplantation.