CRISPR-targeted MAGT1 insertion restores XMEN patient hematopoietic stem cells and lymphocytes
| Autor: | Ezekiel Bello, Marita Bosticardo, Ronald J. Meis, Sherry Koontz, Shengdar Q. Tsai, Juan C Ravell, Michael J. Lenardo, Luigi D. Notarangelo, Benjamin P. Kleinstiver, Xiaolin Wu, Cicera R. Lazzarotto, Harry L. Malech, Guillaume Vayssière, Kennichi C. Dowdell, Colin L. Sweeney, Gary A. Dahl, Taylor Q. Liu, Aaron B. Clark, Uimook Choi, Cristina Corsino, Suk See De Ravin, Siyuan Liu, Julie Brault |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Male
Epstein-Barr Virus Infections Herpesvirus 4 Human Genetic enhancement Immunology Biology X-Linked Combined Immunodeficiency Diseases Biochemistry Mice Inbred NOD medicine Animals CRISPR Humans Lymphocytes Progenitor cell Cation Transport Proteins Cells Cultured Gene Editing Hematopoietic Stem Cell Transplantation Hematopoietic stem cell Genetic Therapy Cell Biology Hematology Gene Therapy Hematopoietic Stem Cells NKG2D Haematopoiesis medicine.anatomical_structure Cancer research Female CRISPR-Cas Systems Stem cell CD8 |
| Zdroj: | Blood |
| Popis: | XMEN disease, defined as “X-linked MAGT1 deficiency with increased susceptibility to Epstein-Barr virus infection and N-linked glycosylation defect,” is a recently described primary immunodeficiency marked by defective T cells and natural killer (NK) cells. Unfortunately, a potentially curative hematopoietic stem cell transplantation is associated with high mortality rates. We sought to develop an ex vivo targeted gene therapy approach for patients with XMEN using a CRISPR/Cas9 adeno-associated vector (AAV) to insert a therapeutic MAGT1 gene at the constitutive locus under the regulation of the endogenous promoter. Clinical translation of CRISPR/Cas9 AAV-targeted gene editing (GE) is hampered by low engraftable gene-edited hematopoietic stem and progenitor cells (HSPCs). Here, we optimized GE conditions by transient enhancement of homology-directed repair while suppressing AAV-associated DNA damage response to achieve highly efficient (>60%) genetic correction in engrafting XMEN HSPCs in transplanted mice. Restored MAGT1 glycosylation function in human NK and CD8+ T cells restored NK group 2 member D (NKG2D) expression and function in XMEN lymphocytes for potential treatment of infections, and it corrected HSPCs for long-term gene therapy, thus offering 2 efficient therapeutic options for XMEN poised for clinical translation. |
| Databáze: | OpenAIRE |
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