The goal of this project is to demonstrate the efficacy of Sleeping Beauty (SB) transposons as vectors for non-viral gene delivery to treat mucopolysaccharidosis (MPS) diseases, specifically Type I, a-iduronidase (IDUA). The project has two sub-goals: 1) Complete experiments that reveal the efficacy of delivery of SB transposons to the mouse to treat a//of the effects of IDUA-deficiency;in particular restoration of IDUA activity in the brain (Aim 1). 2) Reduce potential cytotoxic effects of SB transposon integrants by directing transposition to safe harbors in the mouse genome (Aims 2 - 4).
Aim 1 will complete our assessment ofthe SB system as a vector for gene delivery in the mouse. The next step for development ofthe SB system for humans is to maximize the safety of using transposons that randomly integrate into genomes by directing their integration to sites that are thought to be safe. The principal hvpotheses of this proposal are: 1) Transposons can be delivered directly into the liver and brain for uptake and expression ofthe gene(s) they carry. 2) Certain features of SB Transposons can be exploited to direct their integration into specific sequences in genomes that are considered """"""""safe"""""""" and thereby avoid some ofthe issues associated with viruses used as gene delivery vectors.
The Specific Aims of the proposal are to 1) determine the efficacy of SB transposon delivery to the brain in MPS I mice, 2) develop a 'universal'targeting system to direct SB transposons into selected regions of a DNA molecule, 3) evaluate the ability of a 'universal'targeting system to direct SB transposons to selected regions ofthe mouse genome in vitro, and 4) evaluate the ability of a 'universal'targeting system to direct SB transposons to selected regions ofthe genome in the liver of a living mouse. IMPACT: There are two major impacts of our proposed studies. 1) We will demonstrate that SB transposons can be effectively used to achieve long-term expression in the brain as well as in other tissues and cell types without using viral vectors. 2) By targeting integration of SB transposons to specific sites in the mouse genome, the concerns of insertional mutagenesis will be reduced considerably.

Public Health Relevance

Lysosomal storage disorders are a rare group of inherited diseases caused by genetic deficiency in which patients suffer from skeletal abnormalities, heart and breathing problems, mental retardation and death. It is envisioned in this grant application that one way to treat these diseases would be to restore the missing gene in patients'central nen/ous system (in the brain) to prevent neurodegeneration. Targeting of transposon vectors to specific sites will increase the safety of use of this vector in humans. PROJECT/PERFORIVIANCE SiTE(S) (If additional space is needed, use Project/

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Program Projects (P01)
Project #
5P01HD032652-15
Application #
8380656
Study Section
Special Emphasis Panel (ZHD1-DSR-Z)
Project Start
Project End
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
15
Fiscal Year
2012
Total Cost
$250,274
Indirect Cost
$84,542
Name
University of Minnesota Twin Cities
Department
Type
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Ou, Li; Przybilla, Michael J; Whitley, Chester B (2018) Metabolomics profiling reveals profound metabolic impairments in mice and patients with Sandhoff disease. Mol Genet Metab :
Ou, L; Przybilla, M J; Whitley, C B (2018) SAAMP 2.0: An algorithm to predict genotype-phenotype correlation of lysosomal storage diseases. Clin Genet 93:1008-1014
Aronovich, Elena L; Hyland, Kendra A; Hall, Bryan C et al. (2017) Prolonged Expression of Secreted Enzymes in Dogs After Liver-Directed Delivery of Sleeping Beauty Transposons: Implications for Non-Viral Gene Therapy of Systemic Disease. Hum Gene Ther 28:551-564
Ou, Li; Przybilla, Michael J; Whitley, Chester B (2017) Proteomic analysis of mucopolysaccharidosis I mouse brain with two-dimensional polyacrylamide gel electrophoresis. Mol Genet Metab 120:101-110
Verhaart, Ingrid E C; Robertson, Agata; Wilson, Ian J et al. (2017) Prevalence, incidence and carrier frequency of 5q-linked spinal muscular atrophy - a literature review. Orphanet J Rare Dis 12:124
Ou, Li; Przybilla, Michael J; Whitley, Chester B (2017) Phenotype prediction for mucopolysaccharidosis type I by in silico analysis. Orphanet J Rare Dis 12:125
Hyland, Kendra A; Aronovich, Elena L; Olson, Erik R et al. (2017) Transgene Expression in Dogs After Liver-Directed Hydrodynamic Delivery of Sleeping Beauty Transposons Using Balloon Catheters. Hum Gene Ther 28:541-550
Ou, Li; Przybilla, Michael J; Koniar, Brenda L et al. (2016) Elements of lentiviral vector design toward gene therapy for treating mucopolysaccharidosis I. Mol Genet Metab Rep 8:87-93
Aronovich, Elena L; Hackett, Perry B (2015) Lysosomal storage disease: gene therapy on both sides of the blood-brain barrier. Mol Genet Metab 114:83-93
Satzer, David; DiBartolomeo, Christina; Ritchie, Michael M et al. (2015) Assessment of dysmyelination with RAFFn MRI: application to murine MPS I. PLoS One 10:e0116788

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