Gene replacement of α-globin with β-globin restores hemoglobin balance in β-thalassemia-derived hematopoietic stem and progenitor cells

Cromer, M. K.; Camarena, J.; Martin, R. M.; Lesch, B. J.; Vakulskas, C. A.; Lemgart, V. T.; Zhang, Y.; Goyal, A.; Zhao, F.; Ponce, E.; Srifa, W.; Bak, R. O.; Uchida, N.; Majeti, R.; Sheehan, V. A.; Tisdale, J. F.; Dever, D. P.; Porteus, M.

doi:10.1101/2020.10.28.359315

bioRxiv · 10.1101/2020.10.28.359315

Gene replacement of α-globin with β-globin restores hemoglobin balance in β-thalassemia-derived hematopoietic stem and progenitor cells

Cromer, M. K.·Camarena, J.·Martin, R. M.·Lesch, B. J.·Vakulskas, C. A.·Lemgart, V. T.·Zhang, Y.·Goyal, A.·Zhao, F.·Ponce, E.·Srifa, W.·Bak, R. O.·Uchida, N.·Majeti, R.·Sheehan, V. A.·Tisdale, J. F.·Dever, D. P.·Porteus, M.

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Abstract

{beta}-thalassemia pathology is not only due to loss of {beta}-globin (HBB), but also erythrotoxic accumulation and aggregation of the {beta}-globin binding partner, -globin (HBA1/2). Here we describe a Cas9/AAV6-mediated genome editing strategy that can replace the entire HBA1 gene with a full-length HBB transgene in {beta}-thalassemia-derived hematopoietic stem and progenitor cells (HSPCs), which is sufficient to normalize {beta}-globin:-globin mRNA and protein ratios and restore functional adult hemoglobin tetramers in patient-derived red blood cells. Edited HSPCs were capable of long-term and bi-lineage hematopoietic reconstitution in mice, establishing proof-of-concept for replacement of HBA1 with HBB as a novel therapeutic strategy for curing {beta}-thalassemia.

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Cromer, M. K., Camarena, J., Martin, R. M., Lesch, B. J., Vakulskas, C. A., Lemgart, V. T., Zhang, Y., Goyal, A., Zhao, F., Ponce, E., Srifa, W., Bak, R. O., Uchida, N., Majeti, R., Sheehan, V. A., Tisdale, J. F., Dever, D. P., Porteus, M.. 2020-10-28. Gene replacement of α-globin with β-globin restores hemoglobin balance in β-thalassemia-derived hematopoietic stem and progenitor cells. https://doi.org/10.1101/2020.10.28.359315

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