NHEJ-Mediated Repair of CRISPR-Cas9-Induced DNA Breaks Efficiently Corrects Mutations in HSPCs from Patients with Fanconi Anemia
Por:
Roman-Rodriguez, FJ, Ugalde, L, Alvarez, L, Diez, B, Ramirez, MJ, Risueno, C, Corton, M, Bogliolo, M, Bernal, S, March, F, Ayuso, C, Hanenberg, H, Sevilla, J, Rodriguez-Perales, S, Torres-Ruiz, R, Surralles, J, Bueren, JA, Rio, P
Publicada:
7 nov 2019
Resumen:
Non-homologous end-joining (NHEJ) is the preferred mechanism used by hematopoietic stem cells (HSCs) to repair double-stranded DNA breaks and is particularly increased in cells deficient in the Fanconi anemia (FA) pathway. Here, we show feasible correction of compromised functional phenotypes in hematopoietic cells from multiple FA complementation groups, including FA-A, FA-C, FA-D1, and FA-D2. NHEJ-mediated repair of targeted CRISPR-Cas9-induced DNA breaks generated compensatory insertions and deletions that restore the coding frame of the mutated gene. NHEJ-mediated editing efficacy was initially verified in FA lymphoblastic cell lines and then in primary FA patient-derived CD34(+) cells, which showed marked proliferative advantage and phenotypic correction both in vitro and after transplantation. Importantly, and in contrast to homologous directed repair, NHEJ efficiently targeted primitive human HSCs, indicating that NHEJ editing approaches may constitute a sound alternative for editing self-renewing human HSCs and consequently for treatment of FA and other monogenic diseases affecting the hematopoietic system.
Filiaciones:
Roman-Rodriguez, FJ:
Ctr Invest Energet Medioambientales & Tecnol CIEM, Div Hematopoiet Innovat Therapies, Madrid 28040, Spain
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
Inst Invest Sanitaria Fdn Jimenez Diaz IIS FJD UA, Adv Therapies Unit, Madrid 28040, Spain
Ugalde, L:
Ctr Invest Energet Medioambientales & Tecnol CIEM, Div Hematopoiet Innovat Therapies, Madrid 28040, Spain
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
Inst Invest Sanitaria Fdn Jimenez Diaz IIS FJD UA, Adv Therapies Unit, Madrid 28040, Spain
Alvarez, L:
Ctr Invest Energet Medioambientales & Tecnol CIEM, Div Hematopoiet Innovat Therapies, Madrid 28040, Spain
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
Inst Invest Sanitaria Fdn Jimenez Diaz IIS FJD UA, Adv Therapies Unit, Madrid 28040, Spain
Diez, B:
Ctr Invest Energet Medioambientales & Tecnol CIEM, Div Hematopoiet Innovat Therapies, Madrid 28040, Spain
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
Inst Invest Sanitaria Fdn Jimenez Diaz IIS FJD UA, Adv Therapies Unit, Madrid 28040, Spain
Ramirez, MJ:
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
UAB, Dept Genet & Microbiol, Genome Instabil & DNA Repair Syndromes Grp, Barcelona 08193, Spain
Hosp Santa Creu & Sant Pau, Serv Genet, Barcelona 08025, Spain
Hosp Santa Creu & Sant Pau, Inst Invest Biomed, Barcelona 08025, Spain
Risueno, C:
Ctr Invest Energet Medioambientales & Tecnol CIEM, Div Hematopoiet Innovat Therapies, Madrid 28040, Spain
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
Inst Invest Sanitaria Fdn Jimenez Diaz IIS FJD UA, Adv Therapies Unit, Madrid 28040, Spain
Corton, M:
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
UAM, Hosp Univ Inst Invest Sanitaria Fdn Jimenez Diaz, Dept Genet, Madrid 28040, Spain
Bogliolo, M:
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
UAB, Dept Genet & Microbiol, Genome Instabil & DNA Repair Syndromes Grp, Barcelona 08193, Spain
Hosp Santa Creu & Sant Pau, Serv Genet, Barcelona 08025, Spain
Hosp Santa Creu & Sant Pau, Inst Invest Biomed, Barcelona 08025, Spain
Bernal, S:
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
Hosp Santa Creu & Sant Pau, Serv Genet, Barcelona 08025, Spain
Hosp Santa Creu & Sant Pau, Inst Invest Biomed, Barcelona 08025, Spain
March, F:
Hosp Santa Creu & Sant Pau, Serv Genet, Barcelona 08025, Spain
Hosp Santa Creu & Sant Pau, Inst Invest Biomed, Barcelona 08025, Spain
Ayuso, C:
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
UAM, Hosp Univ Inst Invest Sanitaria Fdn Jimenez Diaz, Dept Genet, Madrid 28040, Spain
Hanenberg, H:
Heinrich Heine Univ, Dept Otorhinolaryngol & Head Neck Surg, D-40225 Dusseldorf, Germany
Univ Duisburg Essen, Univ Childrens Hosp Essen, Dept Pediat 3, D-45122 Essen, Germany
Sevilla, J:
Hosp Univ Nino Jesus, Madrid 28009, Spain
Rodriguez-Perales, S:
CNIO, Mol Cytogenet Grp, Human Canc Genet Program, Madrid 28029, Spain
Torres-Ruiz, R:
CNIO, Mol Cytogenet Grp, Human Canc Genet Program, Madrid 28029, Spain
Univ Barcelona, Sch Med, Josep Carreras Leukemia Res Inst, E-08036 Barcelona, Spain
Univ Barcelona, Sch Med, Dept Biomed, E-08036 Barcelona, Spain
Surralles, J:
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
UAB, Dept Genet & Microbiol, Genome Instabil & DNA Repair Syndromes Grp, Barcelona 08193, Spain
Hosp Santa Creu & Sant Pau, Serv Genet, Barcelona 08025, Spain
Hosp Santa Creu & Sant Pau, Inst Invest Biomed, Barcelona 08025, Spain
Bueren, JA:
Ctr Invest Energet Medioambientales & Tecnol CIEM, Div Hematopoiet Innovat Therapies, Madrid 28040, Spain
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
Inst Invest Sanitaria Fdn Jimenez Diaz IIS FJD UA, Adv Therapies Unit, Madrid 28040, Spain
Rio, P:
Ctr Invest Energet Medioambientales & Tecnol CIEM, Div Hematopoiet Innovat Therapies, Madrid 28040, Spain
Ctr Invest Biomed Red Enfermedades Raras CIBERER, Madrid 28040, Spain
Inst Invest Sanitaria Fdn Jimenez Diaz IIS FJD UA, Adv Therapies Unit, Madrid 28040, Spain
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