RT Journal Article
SR Electronic
T1 Cas9-AAV6 Gene Correction of Beta-Globin in Autologous HSCs Improves Sickle Cell Disease Erythropoiesis in Mice
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.10.13.338319
DO 10.1101/2020.10.13.338319
A1 Adam C. Wilkinson
A1 Daniel P. Dever
A1 Ron Baik
A1 Joab Camarena
A1 Ian Hsu
A1 Carsten T. Charlesworth
A1 Chika Morita
A1 Hiromitsu Nakauchi
A1 Matthew H. Porteus
YR 2020
UL http://biorxiv.org/content/early/2020/10/13/2020.10.13.338319.abstract
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.