PT  - JOURNAL ARTICLE
AU  - Adam C. Wilkinson
AU  - Daniel P. Dever
AU  - Ron Baik
AU  - Joab Camarena
AU  - Ian Hsu
AU  - Carsten T. Charlesworth
AU  - Chika Morita
AU  - Hiromitsu Nakauchi
AU  - Matthew H. Porteus
TI  - Cas9-AAV6 Gene Correction of Beta-Globin in Autologous HSCs Improves Sickle Cell Disease Erythropoiesis in Mice
AID  - 10.1101/2020.10.13.338319
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.10.13.338319
4099  - http://biorxiv.org/content/early/2020/10/13/2020.10.13.338319.short
4100  - http://biorxiv.org/content/early/2020/10/13/2020.10.13.338319.full
AB  - 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 novel paradigm in gene therapy. Although several Cas9-based HBB-correction approaches have been proposed, functional correction of in vivo erythropoiesis has not been investigated. Here, we used 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 observed a direct correlation between increased HBB-corrected myeloid chimerism and normalized in vivo 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.Competing Interest StatementH.N. is a co-founder and shareholder in ReproCELL, Megakaryon and Century Therapeutics. M.H.P. has equity and serves on the scientific advisory board of CRISPR Therapeutics and Allogene Therapeutics. However, none of these companies had input into the design, execution, interpretation, or publication of the work in this manuscript.