In vivo correction of anaemia in β-thalassemic mice by γPNA-mediated gene editing with nanoparticle delivery

• Published in: Nature communications Vol. 7; no. 1; pp. 13304 - 14
• Main Authors: Bahal, Raman, Ali McNeer, Nicole, Quijano, Elias, Liu, Yanfeng, Sulkowski, Parker, Turchick, Audrey, Lu, Yi-Chien, Bhunia, Dinesh C., Manna, Arunava, Greiner, Dale L., Brehm, Michael A., Cheng, Christopher J., López-Giráldez, Francesc, Ricciardi, Adele, Beloor, Jagadish, Krause, Diane S., Kumar, Priti, Gallagher, Patrick G., Braddock, Demetrios T., Mark Saltzman, W., Ly, Danith H., Glazer, Peter M.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 26.10.2016; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 42; 42/41; 631/61/391/2308; 692/4017; Animals; beta-Globins - genetics; beta-Thalassemia - blood; beta-Thalassemia - genetics; beta-Thalassemia - therapy; Cell Line; Disease Models, Animal; DNA - administration & dosage; DNA - genetics; Gene Editing - methods; Genetic Therapy - methods; Hematopoietic Stem Cells - metabolism; Hemoglobins - analysis; Humanities and Social Sciences; Humans; Injections, Intravenous; Mice; Mice, Transgenic; multidisciplinary; Nanoparticles - administration & dosage; Peptide Nucleic Acids - administration & dosage; Peptide Nucleic Acids - genetics; Proto-Oncogene Proteins c-kit - metabolism; Science; Science (multidisciplinary); Stem Cell Factor - administration & dosage; Stem Cell Factor - metabolism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms13304

The blood disorder, β-thalassaemia, is considered an attractive target for gene correction. Site-specific triplex formation has been shown to induce DNA repair and thereby catalyse genome editing. Here we report that triplex-forming peptide nucleic acids (PNAs) substituted at the γ position plus stimulation of the stem cell factor (SCF)/c-Kit pathway yielded high levels of gene editing in haematopoietic stem cells (HSCs) in a mouse model of human β-thalassaemia. Injection of thalassemic mice with SCF plus nanoparticles containing γPNAs and donor DNAs ameliorated the disease phenotype, with sustained elevation of blood haemoglobin levels into the normal range, reduced reticulocytosis, reversal of splenomegaly and up to 7% β-globin gene correction in HSCs, with extremely low off-target effects. The combination of nanoparticle delivery, next generation γPNAs and SCF treatment may offer a minimally invasive treatment for genetic disorders of the blood that can be achieved safely and simply by intravenous administration. Gene editing approaches are widely used for correcting mutations, but their application is largely limited to cells and not living animals. Here the authors show that in vivo γPNA-mediated editing of a β-globin mutation is promoted by SCF and leads to sustained normalization of blood haemoglobin levels β-thalassemic mice.