Dual α-globin-truncated erythropoietin receptor knockin restores hemoglobin production in α-thalassemia-derived erythroid cells

Simon N Chu; Eric Soupene; Devesh Sharma; Roshani Sinha; Travis McCreary; Britney Hernandez; Huifeng Shen; Beeke Wienert; Chance Bowman; Han Yin; Benjamin J Lesch; Kun Jia; Kathleen A Romero; Zachary Kostamo; Yankai Zhang; Tammy Tran; Marco Cordero; Shota Homma; Jessica P Hampton; James M Gardner; Bruce R Conklin; Tippi C MacKenzie; Vivien A Sheehan; Matthew H Porteus; M Kyle Cromer

doi:10.1016/j.celrep.2024.115141

Dual α-globin-truncated erythropoietin receptor knockin restores hemoglobin production in α-thalassemia-derived erythroid cells

Cell Rep. 2025 Jan 3;44(1):115141. doi: 10.1016/j.celrep.2024.115141. Online ahead of print.

Authors

Simon N Chu¹, Eric Soupene², Devesh Sharma³, Roshani Sinha³, Travis McCreary³, Britney Hernandez⁴, Huifeng Shen⁴, Beeke Wienert⁵, Chance Bowman⁶, Han Yin⁷, Benjamin J Lesch⁶, Kun Jia³, Kathleen A Romero⁴, Zachary Kostamo⁴, Yankai Zhang⁴, Tammy Tran³, Marco Cordero³, Shota Homma⁸, Jessica P Hampton⁹, James M Gardner⁷, Bruce R Conklin¹⁰, Tippi C MacKenzie³, Vivien A Sheehan⁴, Matthew H Porteus⁹, M Kyle Cromer¹¹

Affiliations

¹ Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; Eli & Edythe Broad Center for Regeneration Medicine, University of California, San Francisco, San Francisco, CA 94143, USA; Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA.
² Department of Pediatrics, University of California, San Francisco, Oakland, CA 94609, USA.
³ Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; Eli & Edythe Broad Center for Regeneration Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
⁴ Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Emory University School of Medicine, Atlanta, GA 30329, USA.
⁵ Gladstone Institutes, San Francisco, CA 94158, USA.
⁶ Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; Eli & Edythe Broad Center for Regeneration Medicine, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Bioengineering & Therapeutic Sciences, University of California, San Francisco, San Francisco, CA 94158, USA.
⁷ Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA.
⁸ Department of Genetics, Stanford University, Stanford, CA 94305, USA.
⁹ Department of Pediatrics, Stanford University, Stanford, CA 94305, USA.
¹⁰ Gladstone Institutes, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, San Francisco, CA 94158, USA.
¹¹ Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; Eli & Edythe Broad Center for Regeneration Medicine, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Bioengineering & Therapeutic Sciences, University of California, San Francisco, San Francisco, CA 94158, USA. Electronic address: kyle.cromer@uscf.edu.

PMID: 39754719
DOI: 10.1016/j.celrep.2024.115141

Abstract

The most severe form of α-thalassemia results from loss of all four copies of α-globin. Postnatally, patients face challenges similar to β-thalassemia, including severe anemia and erythrotoxicity due to the imbalance of β-globin and α-globin chains. Despite progress in genome editing treatments for β-thalassemia, there is no analogous curative option for α-thalassemia. To address this, we designed a Cas9/AAV6-mediated genome editing strategy that integrates a functional α-globin gene into the β-globin locus in α-thalassemia patient-derived hematopoietic stem and progenitor cells (HSPCs). Incorporation of a truncated erythropoietin receptor transgene into the α-globin integration cassette significantly increased erythropoietic output from edited HSPCs and led to the most robust production of α-globin, and consequently hemoglobin tetramers. By directing edited HSPCs toward increased production of clinically relevant erythroid cells, this approach has the potential to mitigate the limitations of current treatments for the hemoglobinopathies, including low genome editing and low engraftment rates.

Keywords: CP: Stem cell research; CRISPR; RNA sequencing; adeno-associated virus; genome editing; hematopoietic stem and progenitor cells; hemoglobin; red blood cells; α-thalassemia.