title: In vivo correction of anaemia in β-thalassemic mice by γPNA-mediated gene editing with nanoparticle delivery.
abstract: 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.
authors: Department of Therapeutic Radiology, Yale University, New Haven, Connecticut 06520, USA.
authors: Raman
authors: R
authors: Bahal
authors: Department of Therapeutic Radiology, Yale University, New Haven, Connecticut 06520, USA.|Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06511, USA.
authors: Nicole
authors: N
authors: Ali McNeer
authors: Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06511, USA.
authors: Elias
authors: E
authors: Quijano
authors: Department of Therapeutic Radiology, Yale University, New Haven, Connecticut 06520, USA.
authors: Yanfeng
authors: Y
authors: Liu
authors: Department of Therapeutic Radiology, Yale University, New Haven, Connecticut 06520, USA.|Department of Genetics, Yale University, New Haven, Connecticut 06520, USA.
authors: Parker
authors: P
authors: Sulkowski
authors: Department of Therapeutic Radiology, Yale University, New Haven, Connecticut 06520, USA.|Department of Genetics, Yale University, New Haven, Connecticut 06520, USA.
authors: Audrey
authors: A
authors: Turchick
authors: Department of Laboratory Medicine, Yale University, New Haven, Connecticut 06520, USA.
authors: Yi-Chien
authors: YC
authors: Lu
authors: Department of Chemistry and Center for Nucleic Acids Science and Technology (CNAST), Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, USA.
authors: Dinesh C
authors: DC
authors: Bhunia
authors: Department of Chemistry and Center for Nucleic Acids Science and Technology (CNAST), Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, USA.
authors: Arunava
authors: A
authors: Manna
authors: Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
authors: Dale L
authors: DL
authors: Greiner
authors: Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
authors: Michael A
authors: MA
authors: Brehm
authors: Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06511, USA.
authors: Christopher J
authors: CJ
authors: Cheng
authors: Yale Center for Genome Analysis (YCGA), Yale University, Orange, Connecticut 06477, USA.
authors: Francesc
authors: F
authors: López-Giráldez
authors: Department of Therapeutic Radiology, Yale University, New Haven, Connecticut 06520, USA.|Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06511, USA.
authors: Adele
authors: A
authors: Ricciardi
authors: Department of Internal Medicine, Section of Infectious Disease, Yale University, New Haven, Connecticut 06520, USA.
authors: Jagadish
authors: J
authors: Beloor
authors: Department of Laboratory Medicine, Yale University, New Haven, Connecticut 06520, USA.
authors: Diane S
authors: DS
authors: Krause
authors: Department of Internal Medicine, Section of Infectious Disease, Yale University, New Haven, Connecticut 06520, USA.
authors: Priti
authors: P
authors: Kumar
authors: Department of Pediatrics, Yale University, New Haven, Connecticut 06520, USA.
authors: Patrick G
authors: PG
authors: Gallagher
authors: Department of Pathology, Yale University, New Haven, Connecticut 06520, USA.
authors: Demetrios T
authors: DT
authors: Braddock
authors: Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06511, USA.
authors: W
authors: W
authors: Mark Saltzman
authors: Department of Chemistry and Center for Nucleic Acids Science and Technology (CNAST), Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, USA.
authors: Danith H
authors: DH
authors: Ly
authors: Department of Therapeutic Radiology, Yale University, New Haven, Connecticut 06520, USA.|Department of Genetics, Yale University, New Haven, Connecticut 06520, USA.
authors: Peter M
authors: PM
authors: Glazer
mesh_terms: D000818:Animals; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004195:Disease Models, Animal; D000072669:Gene Editing / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006454:Hemoglobins / Q000032:analysis; D006801:Humans; D007275:Injections, Intravenous; D051379:Mice; D008822:Mice, Transgenic; D053758:Nanoparticles / Q000008:administration & dosage; D020135:Peptide Nucleic Acids / Q000008:administration & dosage / Q000235:genetics*; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D019089:Stem Cell Factor / Q000008:administration & dosage / Q000378:metabolism; D055544:beta-Globins / Q000235:genetics; D017086:beta-Thalassemia / Q000097:blood / Q000235:genetics / Q000628:therapy*
chemical_list: D006454:Hemoglobins; D020135:Peptide Nucleic Acids; D019089:Stem Cell Factor; D055544:beta-Globins; D004247:DNA; D019009:Proto-Oncogene Proteins c-kit