title: Adenovirus expressing an NPC1-GFP fusion gene corrects neuronal and nonneuronal defects associated with Niemann pick type C disease.
abstract: Niemann Pick type C (NPC) disease is an autosomal recessive disorder characterized by abnormal cholesterol metabolism and accumulation in lysosomal and endosomal compartments. Although peripheral organs are affected, the progressive neurodegeneration in the brain is typically most deleterious, leading to dystonia, ataxia, seizures, and premature death. Although the two genes underlying this disorder in humans and mouse models of the disease have been identified (NPC1 in 95% and NPC2/HE1 in 5% of human cases), their cellular roles have not Been fully defined, and there is currently no effective treatment for this disorder. To help address these issues, we constructed a recombinant adenovirus, Ad(NPC1-GFP), which contains a cDNA encoding a mouse NPC1 protein with a green fluorescent protein (GFP) fused to its C-terminus. Fluorescence microscopy and cholesterol trafficking assays demonstrate that the GFP-tagged NPC1 protein is functional and detectable in cells from different species (hamster, mouse, human) and of different types (ovary-derived cells, fibroblasts, astrocytes, neurons from peripheral and central nervous systems) in vitro. Combined with results from time-lapse microscopy and in vivo brain injections, our findings suggest that this adenovirus offers advantages for expressing NPC1 and analyzing its cellular localization, movement, functional properties, and beneficial effects in vitro and in vivo.
authors: Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
authors: C A
authors: CA
authors: Paul
authors: P C
authors: PC
authors: Reid
authors: A K
authors: AK
authors: Boegle
authors: B
authors: B
authors: Karten
authors: M
authors: M
authors: Zhang
authors: Z-G
authors: ZG
authors: Jiang
authors: D
authors: D
authors: Franz
authors: L
authors: L
authors: Lin
authors: T Y
authors: TY
authors: Chang
authors: J E
authors: JE
authors: Vance
authors: J
authors: J
authors: Blanchette-Mackie
authors: R A
authors: RA
authors: Maue
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001253:Astrocytes / Q000378:metabolism; D001692:Biological Transport / Q000502:physiology; D001921:Brain / Q000378:metabolism; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D006224:Cricetinae; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000378:metabolism; D000077296:Niemann-Pick C1 Protein; D009542:Niemann-Pick Diseases / Q000235:genetics*; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D002784:Cholesterol