Site-Specific Gene Editing of Human Hematopoietic Stem Cells for X-Linked Hyper-IgM Syndrome

Cell Reports

Xiaoyan Wang¹  Donald B. Kohn²  Beatriz Campo-Fernandez²  Zulema Romero²  Megan D. Hoban²  Alok V. Joglekar²  Aaron R. Cooper²  Joseph D. Long²  Michael L. Kaufman²  Roger P. Hollis²  Georgia R. Lill²  Sorel Fitz-Gibbon³  Caroline Y. Kuo⁴  Satiro de Oliveira⁵ 

[1] Department of General Internal Medicine and Health Services Research, University of California, Los Angeles, Los Angeles, CA 90095, USA;Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA;Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA 90095, USA;Division of Allergy & Immunology, Department of Pediatrics, David Geffen School of Medicine at the University of California, Los Angeles, Los Angeles, CA 90095, USA;Division of Hematology & Oncology, Department of Pediatrics, David Geffen School of Medicine at the University of California, Los Angeles, Los Angeles, CA 90095, USA;

关键词: X-linked hyper-IgM syndrome;    gene editing;    gene therapy;    primary immunodeficiency;    CD40 ligand;    hematopoietic stem cell;    CRISPR/Cas9;    TALEN;   

DOI  :  10.1016/j.celrep.2018.04.103

来源: DOAJ

【 摘 要 】

X-linked hyper-immunoglobulin M (hyper-IgM) syndrome (XHIM) is a primary immunodeficiency due to mutations in CD40 ligand that affect immunoglobulin class-switch recombination and somatic hypermutation. The disease is amenable to gene therapy using retroviral vectors, but dysregulated gene expression results in abnormal lymphoproliferation in mouse models, highlighting the need for alternative strategies. Here, we demonstrate the ability of both the transcription activator-like effector nuclease (TALEN) and clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) platforms to efficiently drive integration of a normal copy of the CD40L cDNA delivered by Adeno-Associated Virus. Site-specific insertion of the donor sequence downstream of the endogenous CD40L promoter maintained physiologic expression of CD40L while overriding all reported downstream mutations. High levels of gene modification were achieved in primary human hematopoietic stem cells (HSCs), as well as in cell lines and XHIM-patient-derived T cells. Notably, gene-corrected HSCs engrafted in immunodeficient mice at clinically relevant frequencies. These studies provide the foundation for a permanent curative therapy in XHIM.

【 授权许可】

Unknown