The broad goal of this research remains the definition of the mechanisms by which acid hydrolases are transported to lysosomes. The program has become progressively more molecular to take advantage of our progress in cloning the human cDNA and genomic sequences specifying beta-glucuronidase, in cloning the rat cDNA for beta-glucuronidase, in cloning the 8.5 kb cDNA for the """"""""215K"""""""" Man 6-P receptor gene, and in discovering a new mouse mutant model for beta-glucuronidase deficiency mucopolysaccharidosis.
The specific aims are: 1) Complete studies defining the genomic organization of the human beta-glucuronidase gene. 2) Define the minimum 5' flanking sequences required for expression of the human beta-glucuronidase gene in mammalian cells. 3) Study the cellular and molecular biology of beta-glucuronidase transport in cultured mammalian cells. 4) Study expression of the human beta-glucuronidase gene in E. coli. 5) Study the expression and transport of the products of the E. coli beta-glucuronidase gene and of E. coli/human beta- glucuronidase chimeric genes in mammalian cells. 6) Study expression and transport of the human beta-glucuronidase gene in transgenic mice. 7) Study the molecular biology and molecular genetics of the cloned """"""""215K"""""""" Man 6-P receptor gene. A variety of biochemical, cell biological. immunological, molecular biological and genetic approaches are applied to the problems of enzyme transport and receptor function. The answers sought have fundamental significance, and may provide information leading to novel therapeutic approaches for lysosomal storage diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM034182-06
Application #
3484785
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1984-06-01
Project End
1993-05-31
Budget Start
1989-06-01
Budget End
1990-05-31
Support Year
6
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Saint Louis University
Department
Type
Schools of Medicine
DUNS #
City
Saint Louis
State
MO
Country
United States
Zip Code
63103
Urayama, Akihiko; Grubb, Jeffrey H; Sly, William S et al. (2016) Pharmacologic manipulation of lysosomal enzyme transport across the blood-brain barrier. J Cereb Blood Flow Metab 36:476-86
Tomatsu, Shunji; Yasuda, Eriko; Patel, Pravin et al. (2014) Morquio A syndrome: diagnosis and current and future therapies. Pediatr Endocrinol Rev 12 Suppl 1:141-51
Sly, William S; Vogler, Carole (2013) The final frontier -- crossing the blood-brain barrier. EMBO Mol Med 5:655-7
Arechederra, Robert L; Waheed, Abdul; Sly, William S et al. (2013) Effect of sulfonamides as carbonic anhydrase VA and VB inhibitors on mitochondrial metabolic energy conversion. Bioorg Med Chem 21:1544-8
Rowan, Daniel J; Tomatsu, Shunji; Grubb, Jeffrey H et al. (2013) Assessment of bone dysplasia by micro-CT and glycosaminoglycan levels in mouse models for mucopolysaccharidosis type I, IIIA, IVA, and VII. J Inherit Metab Dis 36:235-46
Schneider, Hans-Peter; Alt, Marco D; Klier, Michael et al. (2013) GPI-anchored carbonic anhydrase IV displays both intra- and extracellular activity in cRNA-injected oocytes and in mouse neurons. Proc Natl Acad Sci U S A 110:1494-9
Huynh, Ha T; Grubb, Jeffrey H; Vogler, Carole et al. (2012) Biochemical evidence for superior correction of neuronal storage by chemically modified enzyme in murine mucopolysaccharidosis VII. Proc Natl Acad Sci U S A 109:17022-7
McIntyre, Alan; Patiar, Shalini; Wigfield, Simon et al. (2012) Carbonic anhydrase IX promotes tumor growth and necrosis in vivo and inhibition enhances anti-VEGF therapy. Clin Cancer Res 18:3100-11
Makani, Sachin; Chen, Huei-Ying; Esquenazi, Susana et al. (2012) NMDA receptor-dependent afterdepolarizations are curtailed by carbonic anhydrase 14: regulation of a short-term postsynaptic potentiation. J Neurosci 32:16754-62
Rowan, Daniel J; Tomatsu, Shunji; Grubb, Jeffrey H et al. (2012) Long circulating enzyme replacement therapy rescues bone pathology in mucopolysaccharidosis VII murine model. Mol Genet Metab 107:161-72

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