@article{3a283b04b21a4debb89e6dde9dab5786,
title = "Human T cell generation is restored in CD3δ severe combined immunodeficiency through adenine base editing",
abstract = "CD3δ SCID is a devastating inborn error of immunity caused by mutations in CD3D, encoding the invariant CD3δ chain of the CD3/TCR complex necessary for normal thymopoiesis. We demonstrate an adenine base editing (ABE) strategy to restore CD3δ in autologous hematopoietic stem and progenitor cells (HSPCs). Delivery of mRNA encoding a laboratory-evolved ABE and guide RNA into a CD3δ SCID patient's HSPCs resulted in a 71.2% ± 7.85% (n = 3) correction of the pathogenic mutation. Edited HSPCs differentiated in artificial thymic organoids produced mature T cells exhibiting diverse TCR repertoires and TCR-dependent functions. Edited human HSPCs transplanted into immunodeficient mice showed 88% reversion of the CD3D defect in human CD34+ cells isolated from mouse bone marrow after 16 weeks, indicating correction of long-term repopulating HSCs. These findings demonstrate the preclinical efficacy of ABE in HSPCs for the treatment of CD3δ SCID, providing a foundation for the development of a one-time treatment for CD3δ SCID patients.",
keywords = "CD3D severe combined immune deficiency, CITE-seq, artificial thymic organoid, base editing, hematopoietic stem and progenitor cells",
author = "McAuley, {Grace E.} and Gloria Yiu and Chang, {Patrick C.} and Newby, {Gregory A.} and Beatriz Campo-Fernandez and Fitz-Gibbon, {Sorel T.} and Xiaomeng Wu and Kang, {Sung Hae L.} and Amber Garibay and Jeffrey Butler and Valentina Christian and Wong, {Ryan L.} and Everette, {Kelcee A.} and Anthony Azzun and Hila Gelfer and Seet, {Christopher S.} and Aru Narendran and Luis Murguia-Favela and Zulema Romero and Nicola Wright and Liu, {David R.} and Crooks, {Gay M.} and Kohn, {Donald B.}",
note = "Funding Information: These studies were supported by a Translational Research Grant from the Jeffrey Modell Foundation (D.B.K.); NIH T32HL086345 (P.C.C.), NIH U01AI142756 (D.R.L.), NIH RM1 HG009490 (D.R.L.), R35 GM118062 (D.R.L.), the Bill and Melinda Gates Foundation (D.R.L.), the Howard Hughes Medical Institute (D.R.L.), NIH K99 award HL163805 (G.A.N.); NIH K08CA235525 and V Foundation Scholar Award (C.S.S.); and G.Y. is the recipient of the A.P. Giannini Postdoctoral Research Fellowship and Leadership Award. N.W. is supported by the Barb Ibbotson Chair in Pediatric Hematology, Alberta Children's Hospital Research Institute. Core services were supported by the UCLA Technology Center for Bioinformatics and Genomics. Karyotype studies were performed in the UCLA Cytogenetics Laboratory. We would like to thank the UCLA Broad Stem Cell Research Center (BSCRC) Flow Cytometry Core for assistance with FACS sorting. Conceptualization, data curation, formal analysis, investigation, methodology, project administration, visualization, writing (original draft, review & editing), G.E.M. and G.Y.; data curation, investigation, methodology, software, visualization, writing (original draft, review & editing), P.C.C.; investigation, writing (review & editing), G.A.N.; investigation, B.C.-F.; formal analysis, S.T.F.-G.; investigation, X.W.; supervision, writing (review & editing), S.-H.L.K.; investigation, A.G.; investigation, J.B.; investigation, V.C.; methodology, R.L.W.; methodology, K.A.E.; investigation, A.A.; investigation, H.G.; methodology, writing (original draft), C.S.S.; investigation, supervision, A.N.; supervision, L.M.-F.; investigation, Z.R.; resources, writing (original draft), N.W.; resources, supervision, writing (review & editing), D.R.L.; conceptualization, methodology, project administration, supervision, writing (original draft, review & editing), G.M.C.; conceptualization, funding acquisition, methodology, project administration, supervision, writing (original draft, review & editing), D.B.K. G.M.C. and C.S.S. are founders of Pluto Immunotherapeutics Inc. and serve as consultants to this company. D.R.L. is a consultant and equity holder of Prime Medicine, Beam Therapeutics, Pairwise Plants, and Chroma Medicine, companies that use gene editing or genome engineering. We support inclusive, diverse, and equitable conduct of research. We worked to ensure gender balance in the recruitment of human subjects. We worked to ensure diversity in experimental samples through the selection of the cell lines. We worked to ensure diversity in experimental samples through the selection of the genomic datasets. One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in their field of research or within their geographical location. One or more of the authors of this paper self-identifies as a gender minority in their field of research. One or more of the authors of this paper self-identifies as a member of the LGBTQIA+ community. While citing references scientifically relevant for this work, we also actively worked to promote gender balance in our reference list. Funding Information: These studies were supported by a Translational Research Grant from the Jeffrey Modell Foundation (D.B.K.); NIH T32HL086345 (P.C.C.), NIH U01AI142756 (D.R.L.), NIH RM1 HG009490 (D.R.L.), R35 GM118062 (D.R.L.), the Bill and Melinda Gates Foundation (D.R.L.), the Howard Hughes Medical Institute (D.R.L.), NIH K99 award HL163805 (G.A.N.); NIH K08CA235525 and V Foundation Scholar Award (C.S.S.); and G.Y. is the recipient of the A.P. Giannini Postdoctoral Research Fellowship and Leadership Award. N.W. is supported by the Barb Ibbotson Chair in Pediatric Hematology, Alberta Children{\textquoteright}s Hospital Research Institute. Core services were supported by the UCLA Technology Center for Bioinformatics and Genomics. Karyotype studies were performed in the UCLA Cytogenetics Laboratory. We would like to thank the UCLA Broad Stem Cell Research Center (BSCRC) Flow Cytometry Core for assistance with FACS sorting. Publisher Copyright: {\textcopyright} 2023 Elsevier Inc.",
year = "2023",
month = mar,
day = "30",
doi = "10.1016/j.cell.2023.02.027",
language = "English (US)",
volume = "186",
pages = "1398--1416.e23",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "7",
}