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Targeted genome editing in human repopulating haematopoietic stem cells

Author

Listed:
  • Pietro Genovese

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy)

  • Giulia Schiroli

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy
    Vita Salute San Raffaele University, 20132 Milan, Italy)

  • Giulia Escobar

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy
    Vita Salute San Raffaele University, 20132 Milan, Italy)

  • Tiziano Di Tomaso

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy)

  • Claudia Firrito

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy)

  • Andrea Calabria

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy)

  • Davide Moi

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy
    Present address: The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland 4102, Australia.)

  • Roberta Mazzieri

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy
    Present address: The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland 4102, Australia.)

  • Chiara Bonini

    (Experimental Hematology Unit, San Raffaele Scientific Institute, 20132 Milan, Italy)

  • Michael C. Holmes

    (Sangamo BioSciences Inc.)

  • Philip D. Gregory

    (Sangamo BioSciences Inc.)

  • Mirjam van der Burg

    (University Medical Center, 3015 Rotterdam, The Netherlands)

  • Bernhard Gentner

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy
    Vita Salute San Raffaele University, 20132 Milan, Italy)

  • Eugenio Montini

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy)

  • Angelo Lombardo

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy
    Vita Salute San Raffaele University, 20132 Milan, Italy)

  • Luigi Naldini

    (TIGET, San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, 20132 Milan, Italy
    Vita Salute San Raffaele University, 20132 Milan, Italy)

Abstract

Targeted genome editing by artificial nucleases has brought the goal of site-specific transgene integration and gene correction within the reach of gene therapy. However, its application to long-term repopulating haematopoietic stem cells (HSCs) has remained elusive. Here we show that poor permissiveness to gene transfer and limited proficiency of the homology-directed DNA repair pathway constrain gene targeting in human HSCs. By tailoring delivery platforms and culture conditions we overcame these barriers and provide stringent evidence of targeted integration in human HSCs by long-term multilineage repopulation of transplanted mice. We demonstrate the therapeutic potential of our strategy by targeting a corrective complementary DNA into the IL2RG gene of HSCs from healthy donors and a subject with X-linked severe combined immunodeficiency (SCID-X1). Gene-edited HSCs sustained normal haematopoiesis and gave rise to functional lymphoid cells that possess a selective growth advantage over those carrying disruptive IL2RG mutations. These results open up new avenues for treating SCID-X1 and other diseases.

Suggested Citation

  • Pietro Genovese & Giulia Schiroli & Giulia Escobar & Tiziano Di Tomaso & Claudia Firrito & Andrea Calabria & Davide Moi & Roberta Mazzieri & Chiara Bonini & Michael C. Holmes & Philip D. Gregory & Mir, 2014. "Targeted genome editing in human repopulating haematopoietic stem cells," Nature, Nature, vol. 510(7504), pages 235-240, June.
    • Handle: RePEc:nat:nature:v:510:y:2014:i:7504:d:10.1038_nature13420
    DOI: 10.1038/nature13420
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    Citations

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    Cited by:

    1. Ron Baik & M. Kyle Cromer & Steve E. Glenn & Christopher A. Vakulskas & Kay O. Chmielewski & Amanda M. Dudek & William N. Feist & Julia Klermund & Suzette Shipp & Toni Cathomen & Daniel P. Dever & Mat, 2024. "Transient inhibition of 53BP1 increases the frequency of targeted integration in human hematopoietic stem and progenitor cells," Nature Communications, Nature, vol. 15(1), pages 1-14, December.
    2. Daniel T. Claiborne & Zachary Detwiler & Steffen S. Docken & Todd D. Borland & Deborah Cromer & Amanda Simkhovich & Youdiil Ophinni & Ken Okawa & Timothy Bateson & Tao Chen & Wesley Hudson & Radiana T, 2025. "High frequency CCR5 editing in human hematopoietic stem progenitor cells protects xenograft mice from HIV infection," Nature Communications, Nature, vol. 16(1), pages 1-14, December.
    3. Sebastian M. Siegner & Laura Ugalde & Alexandra Clemens & Laura Garcia-Garcia & Juan A. Bueren & Paula Rio & Mehmet E. Karasu & Jacob E. Corn, 2022. "Adenine base editing efficiently restores the function of Fanconi anemia hematopoietic stem and progenitor cells," Nature Communications, Nature, vol. 13(1), pages 1-15, December.
    4. Florence Borot & Olivier Humbert & Jeffrey T. Ehmsen & Emily Fields & Sajeev Kohli & Stefan Radtke & Kyle Swing & Dnyanada Pande & Mark R. Enstrom & George S. Laszlo & Thiyagaraj Mayuranathan & Abdull, 2025. "Multiplex base editing to protect from CD33 directed drugs for immune and gene therapy," Nature Communications, Nature, vol. 16(1), pages 1-17, December.
    5. Jonas Holst Wolff & Thomas Wisbech Skov & Didde Haslund & Sofie Rahbek Dorset & Anne Louise S. Revenfeld & Clotilde Aussel & Sofie E. Jørgensen & Mette Holm & Martin K. Thomsen & Sandra Ammann & Toni , 2025. "Targeted gene editing and near-universal cDNA insertion of CYBA and CYBB as a treatment for chronic granulomatous disease," Nature Communications, Nature, vol. 16(1), pages 1-16, December.
    6. William N. Feist & Sofia E. Luna & Kaya Ben-Efraim & Maria V. Filsinger Interrante & Alvaro Amorin & Nicole M. Johnston & Theodora U. J. Bruun & Ashley Utz & Hana Y. Ghanim & Benjamin J. Lesch & There, 2025. "Multilayered HIV-1 resistance in HSPCs through CCR5 Knockout and B cell secretion of HIV-inhibiting antibodies," Nature Communications, Nature, vol. 16(1), pages 1-18, December.

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