Mucopolysaccharidosis III B (Sanfilippo type 13, MPS IIIB) is a lysosomal storage disorder due to the inherited deficiency of a-N-acetylglucosaminidase (NaGlu). The disease is characterized by mild somatic disease with severe neurological degeneration in most of patients by 6-10 years of age with rapid and progressive deterioration of social and adaptive abilities leading to premature death. No treatment is currently available for the central nervous system (CNS) disorder of MPS III B patients, which is usually the cause of premature death. Adeno-associated virus (AAV) has been shown to provide a promising gene delivery system for its ability of infecting wide range of tissues/organs and with no known pathogenesis in human. In this project, AAV-mediated gene therapy for the CNS disease of MPS III B is to be studied using a knock-out mouse model. Two AAV-vectors, containing human NaGlu cDNA driven by a CW promoter or the endogenous brain promoter, neuron specific enolase (NSE) promoter, have been constructed and have shown efficient expression of functional NaGlu and the correction of lysosomal storage by recombinant NaGlu in vitro in MPS IIIB cell cultures and in vivo in mouse brain. The vectors are to be delivered into multiple brain areas of MPS III B mice by direct microinjection, to study more efficient in vivo expression and distribution of rNaGlu and the correction of lysosomal storage in the brain after the injection. In addition, studies will be conducted to develop more efficient means of gene delivery into brain because limited distribution of gene therapy vectors in brain is one of the biggest obstacles for CNS therapies. It will be achieved by delivering AAV vectors containing an enhanced green fluorescent protein gene into multiple mouse brain areas by direct microinjection, to a) compare the distribution of gene expression mediated by different AAV serotype (1, 2 and 5) vectors; b) study the dispersion of gene expression delivered into mouse brain through different injection route, especially peripheral delivery; and c) study the spread of AAV vectors by modifying the delivery media of AAV viral vectors; d). Conduct multiple site infusion by a single injection, to increase the distribution of AAV vectors after a single injection. The goal of this project is to study the feasibility of using AAV-mediated gene therapy to treat the CNS disease in MPS III B children and to achieve broad distribution of AAV vectors in CNS, using mouse model. The long-term goal of this project is to develop novel therapy for CNS disease in MPS IIIB patients.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
1R01DK063972-01
Application #
6547921
Study Section
Special Emphasis Panel (ZNS1-SRB-S (01))
Program Officer
Mckeon, Catherine T
Project Start
2002-08-15
Project End
2005-06-30
Budget Start
2002-08-15
Budget End
2003-06-30
Support Year
1
Fiscal Year
2002
Total Cost
$252,350
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Pediatrics
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Jaimes, Edgar A; Hua, Ping; Tian, Run-Xia et al. (2010) Human glomerular endothelium: interplay among glucose, free fatty acids, angiotensin II, and oxidative stress. Am J Physiol Renal Physiol 298:F125-32
Fu, Haiyan; DiRosario, Julianne; Kang, Lu et al. (2010) Restoration of central nervous system alpha-N-acetylglucosaminidase activity and therapeutic benefits in mucopolysaccharidosis IIIB mice by a single intracisternal recombinant adeno-associated viral type 2 vector delivery. J Gene Med 12:624-33
McCarty, D M; DiRosario, J; Gulaid, K et al. (2009) Mannitol-facilitated CNS entry of rAAV2 vector significantly delayed the neurological disease progression in MPS IIIB mice. Gene Ther 16:1340-52
Pearse, Damien D; Tian, Run-Xia; Nigro, Jessica et al. (2008) Angiotensin II increases the expression of the transcription factor ETS-1 in mesangial cells. Am J Physiol Renal Physiol 294:F1094-100
Jaimes, Edgar A; Zhou, Ming-Sheng; Pearse, Damien D et al. (2008) Upregulation of cortical COX-2 in salt-sensitive hypertension: role of angiotensin II and reactive oxygen species. Am J Physiol Renal Physiol 294:F385-92
Fu, H; Kang, L; Jennings, J S et al. (2007) Significantly increased lifespan and improved behavioral performances by rAAV gene delivery in adult mucopolysaccharidosis IIIB mice. Gene Ther 14:1065-77
Fu, Haiyan; Muenzer, Joseph; Samulski, Richard J et al. (2003) Self-complementary adeno-associated virus serotype 2 vector: global distribution and broad dispersion of AAV-mediated transgene expression in mouse brain. Mol Ther 8:911-7