{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:20:08Z","timestamp":1792790408374,"version":"4.5.0"},"posted":{"date-parts":[[2026,9,17]]},"group-title":"Bioengineering","reference-count":0,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T00:00:00Z","timestamp":1789603200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institutes of Health, United States","award":["R01HL130040"],"award-info":[{"award-number":["R01HL130040"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"crossref"}]},{"award":["INV-017692"],"award-info":[{"award-number":["INV-017692"]}],"id":[{"id":"https:\/\/ror.org\/0456r8d26","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,17]]},"abstract":"<jats:p>In vivo hematopoietic stem\/progenitor cell (HSPC) gene therapy remains limited by low gene-editing efficiency and a lack of clinically applicable selection strategies to enrich therapeutically corrected progeny. We used in vivo base and prime editing to introduce a nonpathogenic EPOR variant into HSPCs, conferring erythropoietin hypersensitivity and promoting preferential expansion of gene-corrected erythroid cells. EPOR editing was combined with three therapeutic approaches for the correction of hemoglobinopathies: \u03b3-globin gene addition, \u03b3-globin reactivation, or correction of the sickle cell disease mutation. Tropism-modified helper-dependent adenoviral vectors (HDAd6\/3+) targeting HSPCs were used to simultaneously deliver the EPOR-editing machinery and the corresponding therapeutic components. In vitro studies in an erythroid progenitor cell line and primary CD34+ cells demonstrated that the EPORW439* variant conferred a strong proliferative advantage to therapeutically modified erythroid progenitors. Mice humanized with CD34+ cells from a \u03b2\u2070\/\u03b2\u2070-thalassemia patient were subjected to EPORW439*-mediated EPO hypersensitivity alongside a therapeutic \u03b3-globin transgene which resulted in &gt;70% HbF-positive erythroid cells and substantial reversion of the disease-associated phenotype, including reduced oxidative stress, near-complete elimination of splenic iron deposition, and reduced splenomegaly. Importantly, these effects were achieved after simple intravenous administration of the vectors following HSPC mobilization and cytokine prophylaxis, without subsequent pharmacologic selection. Together, these findings establish a strategy to amplify the therapeutic benefit of otherwise limited in vivo HSPC gene editing for hemoglobinopathies.<\/jats:p>","DOI":"10.64898\/2026.09.15.751289","type":"posted-content","created":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T04:35:27Z","timestamp":1789706127000},"source":"Crossref","is-referenced-by-count":0,"title":["In vivo HSPC gene therapy of hemoglobinopathies without drug selection of corrected cells"],"prefix":"10.64898","author":[{"given":"Akshara","family":"Sakunthala Velmurugan","sequence":"first","affiliation":[{"name":"University of Washington, Department of Medicine, Division of Medical Genetics, Box 357720, Seattle, WA 98195, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-2378-1442","authenticated-orcid":false,"given":"Kiriaki","family":"Paschoudi","sequence":"additional","affiliation":[{"name":"Gene and Cell Therapy Center, Hematology Department, George Papanicolaou Hospital, Thessaloniki, Greece;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1057-3892","authenticated-orcid":false,"given":"Jack A.","family":"Queenan","sequence":"additional","affiliation":[{"name":"Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Department of Chemistry and Chemical Biology, Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sucheol","family":"Gil","sequence":"additional","affiliation":[{"name":"University of Washington, Department of Medicine, Division of Medical Genetics, Box 357720, Seattle, WA 98195, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aidan","family":"Maynard","sequence":"additional","affiliation":[{"name":"University of Washington, Department of Medicine, Division of Medical Genetics, Box 357720, Seattle, WA 98195, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charlie E","family":"Fontana","sequence":"additional","affiliation":[{"name":"University of Washington, Department of Medicine, Division of Medical Genetics, Box 357720, Seattle, WA 98195, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4381-0989","authenticated-orcid":false,"given":"Reva","family":"Sharma","sequence":"additional","affiliation":[{"name":"University of Washington, Department of Medicine, Division of Medical Genetics, Box 357720, Seattle, WA 98195, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1364-1266","authenticated-orcid":false,"given":"Anika","family":"Long","sequence":"additional","affiliation":[{"name":"University of Washington, Department of Medicine, Division of Medical Genetics, Box 357720, Seattle, WA 98195, 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Thessaloniki;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2043-9630","authenticated-orcid":false,"given":"Efthymia","family":"Vlachaki","sequence":"additional","affiliation":[{"name":"Second Department of Internal Medicine, School of Medicine, Aristotle University of Thessaloniki;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nouraiz","family":"Ahmed","sequence":"additional","affiliation":[{"name":"Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Department of Chemistry and Chemical Biology, Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander A","family":"Sousa","sequence":"additional","affiliation":[{"name":"Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and Harvard, Department of Chemistry and Chemical Biology, Howard Hughes Medical 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