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. Scott McIvor

Research output: Contribution to journalArticlepeer-review

42 Scopus citations

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

Bibliographical note

Funding Information:
We are grateful to Dr. Joseph Muenzer for providing the MPS II mouse strain. The authors would like to thank Carolyn Gaspar, Annemarie Ledeboer, Melanie Butler, Marshall Huston, and Amy Manning-Bog for assistance in necropsy and figure preparation and also Seventh Wave Laboratories (Maryland Heights, MO) for performing necropsy and histopathology evaluation (Mark Martinez, DVM, DACVP) and Pacific BioLabs (Hercules, CA) for performing mass spectrometry analysis of dermatan and heparan sulfate levels (Rick Staub and Vy Tran). This work was supported by Sangamo Therapeutics. The materials described herein will be provided upon request after execution of a material transfer agreement with the University of Minnesota and/or Sangamo Therapeutics, as appropriate.

Funding Information:
We are grateful to Dr. Joseph Muenzer for providing the MPS II mouse strain. The authors would like to thank Carolyn Gaspar, Annemarie Ledeboer, Melanie Butler, Marshall Huston, and Amy Manning-Bog for assistance in necropsy and figure preparation and also Seventh Wave Laboratories (Maryland Heights, MO) for performing necropsy and histopathology evaluation (Mark Martinez, DVM, DACVP) and Pacific BioLabs (Hercules, CA) for performing mass spectrometry analysis of dermatan and heparan sulfate levels (Rick Staub and Vy Tran). This work was supported by Sangamo Therapeutics . The materials described herein will be provided upon request after execution of a material transfer agreement with the University of Minnesota and/or Sangamo Therapeutics, as appropriate.

Publisher Copyright:
© 2018 The Authors

Keywords

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

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