Dose-Dependent Prevention of Metabolic and Neurologic Disease in Murine MPS II by ZFN-Mediated In Vivo Genome Editing

Kanut Laoharawee, Russell C. DeKelver, Kelly M. Podetz-Pedersen, Michelle Rohde, Scott Sproul, Hoang Oanh Nguyen, Tam Nguyen, Susan J. St. Martin, Li Ou, Susan Tom, Robert Radeke, Kathleen E. Meyer, Michael C. Holmes, Chester B Whitley, Thomas Wechsler, R S Mc Ivor

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Mucopolysaccharidosis type II (MPS II) is an X-linked recessive lysosomal disorder caused by deficiency of iduronate 2-sulfatase (IDS), leading to accumulation of glycosaminoglycans (GAGs) in tissues of affected individuals, progressive disease, and shortened lifespan. Currently available enzyme replacement therapy (ERT) requires lifelong infusions and does not provide neurologic benefit. We utilized a zinc finger nuclease (ZFN)-targeting system to mediate genome editing for insertion of the human IDS (hIDS) coding sequence into a “safe harbor” site, intron 1 of the albumin locus in hepatocytes of an MPS II mouse model. Three dose levels of recombinant AAV2/8 vectors encoding a pair of ZFNs and a hIDS cDNA donor were administered systemically in MPS II mice. Supraphysiological, vector dose-dependent levels of IDS enzyme were observed in the circulation and peripheral organs of ZFN+donor-treated mice. GAG contents were markedly reduced in tissues from all ZFN+donor-treated groups. Surprisingly, we also demonstrate that ZFN-mediated genome editing prevented the development of neurocognitive deficit in young MPS II mice (6–9 weeks old) treated at high vector dose levels. We conclude that this ZFN-based platform for expression of therapeutic proteins from the albumin locus is a promising approach for treatment of MPS II and other lysosomal diseases. AAV-mediated in vivo delivery of ZFN and IDS donor resulted in site-specific gene insertion and dose-dependent IDS expression in a mouse model of MPS II. These results support a currently open clinical trial, the first ever in vivo human genome editing study to be conducted.

Original languageEnglish (US)
Pages (from-to)1127-1136
Number of pages10
JournalMolecular Therapy
Volume26
Issue number4
DOIs
StatePublished - Apr 4 2018

Fingerprint

Mucopolysaccharidosis II
Zinc Fingers
Metabolic Diseases
Nervous System Diseases
Iduronate Sulfatase
Glycosaminoglycans
Albumins
Mucopolysaccharidosis I
Enzyme Replacement Therapy
Insertional Mutagenesis
Human Genome
Gene Editing
Introns
Nervous System
Hepatocytes
Complementary DNA
Clinical Trials
Enzymes
Therapeutics

Keywords

  • Hunter syndrome
  • MPS II
  • albumin locus
  • gene therapy
  • iduronate 2-sulfatase
  • in vivo genome editing
  • lysosomal disease
  • zinc finger nuclease

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Dose-Dependent Prevention of Metabolic and Neurologic Disease in Murine MPS II by ZFN-Mediated In Vivo Genome Editing. / Laoharawee, Kanut; DeKelver, Russell C.; Podetz-Pedersen, Kelly M.; Rohde, Michelle; Sproul, Scott; Nguyen, Hoang Oanh; Nguyen, Tam; St. Martin, Susan J.; Ou, Li; Tom, Susan; Radeke, Robert; Meyer, Kathleen E.; Holmes, Michael C.; Whitley, Chester B; Wechsler, Thomas; Mc Ivor, R S.

In: Molecular Therapy, Vol. 26, No. 4, 04.04.2018, p. 1127-1136.

Research output: Contribution to journalArticle

Laoharawee, K, DeKelver, RC, Podetz-Pedersen, KM, Rohde, M, Sproul, S, Nguyen, HO, Nguyen, T, St. Martin, SJ, Ou, L, Tom, S, Radeke, R, Meyer, KE, Holmes, MC, Whitley, CB, Wechsler, T & Mc Ivor, RS 2018, 'Dose-Dependent Prevention of Metabolic and Neurologic Disease in Murine MPS II by ZFN-Mediated In Vivo Genome Editing', Molecular Therapy, vol. 26, no. 4, pp. 1127-1136. https://doi.org/10.1016/j.ymthe.2018.03.002
Laoharawee, Kanut ; DeKelver, Russell C. ; Podetz-Pedersen, Kelly M. ; Rohde, Michelle ; Sproul, Scott ; Nguyen, Hoang Oanh ; Nguyen, Tam ; St. Martin, Susan J. ; Ou, Li ; Tom, Susan ; Radeke, Robert ; Meyer, Kathleen E. ; Holmes, Michael C. ; Whitley, Chester B ; Wechsler, Thomas ; Mc Ivor, R S. / Dose-Dependent Prevention of Metabolic and Neurologic Disease in Murine MPS II by ZFN-Mediated In Vivo Genome Editing. In: Molecular Therapy. 2018 ; Vol. 26, No. 4. pp. 1127-1136.
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abstract = "Mucopolysaccharidosis type II (MPS II) is an X-linked recessive lysosomal disorder caused by deficiency of iduronate 2-sulfatase (IDS), leading to accumulation of glycosaminoglycans (GAGs) in tissues of affected individuals, progressive disease, and shortened lifespan. Currently available enzyme replacement therapy (ERT) requires lifelong infusions and does not provide neurologic benefit. We utilized a zinc finger nuclease (ZFN)-targeting system to mediate genome editing for insertion of the human IDS (hIDS) coding sequence into a “safe harbor” site, intron 1 of the albumin locus in hepatocytes of an MPS II mouse model. Three dose levels of recombinant AAV2/8 vectors encoding a pair of ZFNs and a hIDS cDNA donor were administered systemically in MPS II mice. Supraphysiological, vector dose-dependent levels of IDS enzyme were observed in the circulation and peripheral organs of ZFN+donor-treated mice. GAG contents were markedly reduced in tissues from all ZFN+donor-treated groups. Surprisingly, we also demonstrate that ZFN-mediated genome editing prevented the development of neurocognitive deficit in young MPS II mice (6–9 weeks old) treated at high vector dose levels. We conclude that this ZFN-based platform for expression of therapeutic proteins from the albumin locus is a promising approach for treatment of MPS II and other lysosomal diseases. AAV-mediated in vivo delivery of ZFN and IDS donor resulted in site-specific gene insertion and dose-dependent IDS expression in a mouse model of MPS II. These results support a currently open clinical trial, the first ever in vivo human genome editing study to be conducted.",
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