Humans ; alpha-Synuclein/genetics* ; Parkinson Disease/pathology*

Genetic Markers ; Genome-Wide Association Study ; HLA-DQ Antigens ; genetics ; HLA-DR Antigens ; genetics ; Humans ; Inflammation ; complications ; genetics ; pathology ; Parkinson Disease ; diagnosis ; genetics ; pathology ; Risk Factors ; Time Factors

The generation of disease-specific induced pluripotent stem cell (iPSC) lines from patients with incurable diseases is a promising approach for studying disease mechanisms and drug screening. Such innovation enables to obtain autologous cell sources in regenerative medicine. Herein, we report the generation and characterization of iPSCs from fibroblasts of patients with sporadic or familial diseases, including Parkinson's disease (PD), Alzheimer's disease (AD), juvenile-onset, type I diabetes mellitus (JDM), and Duchenne type muscular dystrophy (DMD), as well as from normal human fibroblasts (WT). As an example to modeling disease using disease-specific iPSCs, we also discuss the previously established childhood cerebral adrenoleukodystrophy (CCALD)- and adrenomyeloneuropathy (AMN)-iPSCs by our group. Through DNA fingerprinting analysis, the origins of generated disease-specific iPSC lines were identified. Each iPSC line exhibited an intense alkaline phosphatase activity, expression of pluripotent markers, and the potential to differentiate into all three embryonic germ layers: the ectoderm, endoderm, and mesoderm. Expression of endogenous pluripotent markers and downregulation of retrovirus-delivered transgenes [OCT4 (POU5F1), SOX2, KLF4, and c-MYC] were observed in the generated iPSCs. Collectively, our results demonstrated that disease-specific iPSC lines characteristically resembled hESC lines. Furthermore, we were able to differentiate PD-iPSCs, one of the disease-specific-iPSC lines we generated, into dopaminergic (DA) neurons, the cell type mostly affected by PD. These PD-specific DA neurons along with other examples of cell models derived from disease-specific iPSCs would provide a powerful platform for examining the pathophysiology of relevant diseases at the cellular and molecular levels and for developing new drugs and therapeutic regimens.
Alzheimer Disease/genetics/*pathology ; Cell Differentiation ; Cells, Cultured ; Diabetes Mellitus, Type 1/genetics/*pathology ; Drug Discovery/*methods ; Fibroblasts/cytology/metabolism/pathology ; Gene Expression ; Humans ; Induced Pluripotent Stem Cells/cytology/metabolism/*pathology ; Muscular Dystrophy, Duchenne/genetics/*pathology ; Parkinson Disease/genetics/*pathology

The leucine-rich repeat kinase2 (LRRK2) has been identified to be the gene causing autosomal dominant inherited Parkinson's disease(PD)8. The clinical features of this type of PD are similar to those of idiopathic PD, but the pathological changes are diverse. The mutation types and frequencies of the LRRK2 distribute unevenly in different populations. LRRK2 is a large complex protein with multiple functions and expresses widely in human body. Sequence alignment shows that LRRK2 might be a multiple function kinase for substrate phosphorylation and might also act as a scaffolding protein. Further study on the physiological function and pathogenic mechanism of LRRK2 will help to find out the possible pathogenesis and new treatment for PD.
Animals ; Continental Population Groups ; genetics ; Humans ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 ; Mutation ; Parkinson Disease ; genetics ; pathology ; Protein-Serine-Threonine Kinases ; chemistry ; genetics ; metabolism ; Sequence Alignment

Brain ; pathology ; Genome-Wide Association Study ; Humans ; Parkinson Disease ; genetics ; Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases ; genetics ; Proto-Oncogene Proteins c-crk ; genetics

Cholesterol is an essential component for neuronal physiology not only during development stage but also in the adult life. Cholesterol metabolism in brain is independent from that in peripheral tissues due to blood-brain barrier. The content of cholesterol in brain must be accurately maintained in order to keep brain function well. Defects in brain cholesterol metabolism has been shown to be implicated in neurodegenerative diseases, such as Alzheimer's disease (AD), Huntington's disease (HD), Parkinson's disease (PD), and some cognitive deficits typical of the old age. The brain contains large amount of cholesterol, but the cholesterol metabolism and its complex homeostasis regulation are currently poorly understood. This review will seek to integrate current knowledge about the brain cholesterol metabolism with molecular mechanisms.
ATP-Binding Cassette Transporters ; genetics ; metabolism ; Alzheimer Disease ; genetics ; metabolism ; pathology ; Blood-Brain Barrier ; Brain ; metabolism ; pathology ; Cholesterol ; metabolism ; Gene Expression Regulation ; Homeostasis ; Humans ; Huntington Disease ; genetics ; metabolism ; pathology ; Hydroxycholesterols ; metabolism ; Lipid Metabolism ; genetics ; Neurons ; metabolism ; pathology ; Parkinson Disease ; genetics ; metabolism ; pathology ; Receptors, Lipoprotein ; genetics ; metabolism

Parkinson's disease (PD) is an age-related progressive neurodegenerative disease associated with selective loss of dopaminergic neurons. The characteristic hallmark of the disease is intracytoplasmic proteinacious inclusion bodies called Lewy bodies, primarily consisting of a presynaptic protein alpha-synuclein. Oxidative stress-mediated damage to macromolecules have been shown to occur frequently in PD. Oxidative damage to DNA in the form of oxidized guanine (8-oxodG) accumulates in both the mitochondrial and nuclear DNA of dopaminergic neurons of the substantia nigra in PD. 8-oxodG-mediated transcriptional mutagenesis has been shown to have the potential to alter phenotype of cells through production of mutant pool of proteins. This review comprehensively summarizes the role of oxidative stress-mediated damage incurred during neurodegeneration, and highlights the scope of transcriptional mutagenesis event in leading to alpha-synuclein aggregation as seen in PD.
Amino Acid Sequence ; Animals ; Deoxyguanosine/*analogs & derivatives/metabolism ; Humans ; Molecular Sequence Data ; Mutagenesis ; *Oxidative Stress ; Parkinson Disease/*genetics/metabolism/pathology ; Protein Aggregation, Pathological/*genetics/metabolism/pathology ; Substantia Nigra/metabolism/*pathology ; Transcription, Genetic ; alpha-Synuclein/chemistry/*genetics

OBJECTIVETo analyze the clinical and genetic features of a family with Parkinson's disease caused by expansion of CAG triplet repeat in the ATXN2 gene.

METHODSThe CAG/CAA repeat in the ATXN2 gene was analyzed by polymerase chain reaction (PCR) and Sanger sequencing.

RESULTSMolecular testing has documented a pathological heterozygous expansion of the CAG repeat from 33 to 35 in 6 patients and other 8 family members. Two patients had pure CAG triplet repeat expansion in their ATXN2 gene, while others had CAA interruption.

CONCLUSIONExpanded CAG/CAA repeat in the ATXN2 gene is the causative mutation of the disease in this family.The 8 members with expanded CAG/CAA repeat may be asymptomatic patients. It is supposed that the number and configuration of the ATXN2 CAG/CAA repeat expansion may play an important role in the phenotypic variability of Parkinson's disease.

Aged ; Ataxin-2 ; genetics ; Base Sequence ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; pathology ; Pedigree ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; methods ; Trinucleotide Repeat Expansion ; genetics

OBJECTIVETo silence the expression of alpha-synuclein in MN9D dopaminergic cells using vector mediated RNA interference (RNAi) and examined its effects on cell proliferation and viability.

METHODSWe identified two 19-nucleotide stretches within the coding region of the alpha-synuclein gene and designed three sets of oligonucleotides to generate double-stranded (ds) oligos. The ds oligos were inserted into the pENTR/H1/TO vector and transfected into MN9D dopaminergic cells. alpha-Synuclein expression was detected by RT-PCR, real-time PCR, immunocytochemistry staining and Western blot. In addition, we measured cell proliferation using growth curves and cell viability by 3-(4, 5)-dimethylthiahiazo (-z-y1)-3, 5-di- phenytetrazoliumromide (MTT).

RESULTSThe mRNA and protein levels of alpha-synuclein gene were significantly down-regulated in pSH2/alpha-SYN-transfected cells compared with control MN9D and pSH/CON-transfected MN9D cells, while pSH1/alpha-SYN-transfected cells showed no significant difference. Silencing alpha-synuclein expression does not affect cell proliferation but may decrease cell viability.

CONCLUSIONOur results demonstrated pSH2/alpha-SYN is an effective small interfering RNA (siRNA) sequence and potent silencing of mouse alpha-synuclein expression in MN9D cells by vector-based RNAi, which provides the tools for studying the normal function of alpha-synuclein and examining its role in Parkinson's disease (PD) pathogenesis. alpha-Synuclein may be important for the viability of MN9D cells, and loss of alpha-synuclein may induce cell injury directly or indirectly.

Animals ; Cell Line ; Cell Proliferation ; Cell Survival ; drug effects ; genetics ; Down-Regulation ; drug effects ; genetics ; Gene Silencing ; Genetic Vectors ; genetics ; Hybridomas ; Mice ; Mice, Inbred C57BL ; Nerve Degeneration ; genetics ; metabolism ; Neurons ; drug effects ; metabolism ; pathology ; Oligonucleotides ; genetics ; Parkinson Disease ; genetics ; metabolism ; Plasmids ; genetics ; RNA Interference ; RNA, Double-Stranded ; genetics ; pharmacology ; RNA, Small Interfering ; genetics ; Transfection ; methods ; alpha-Synuclein ; genetics ; metabolism

Many major neurodegenerative diseases are associated with proteins misfolding and aggregation, which are also called "neurodegenerative conformational disease". The interaction of gene mutation and environmental factors are probably primary events resulting in oligomer and aggregate formations of proteins. Moreover, the dysfunctions of protein control systems, i.e. the ubiquitin-proteasome system and autophagy-lysosomal system, also contribute to the neurodegenerative process. The present review mainly summarizes protein misfolding and aggregation in the development of neurodegenerative conformational disease and the underling mechanisms, as well as upregulation of heatshock proteins as a promising treatment method for this kind of disease.
Alzheimer Disease ; drug therapy ; genetics ; metabolism ; pathology ; Animals ; Annona ; chemistry ; Autophagy ; Benzeneacetamides ; isolation & purification ; therapeutic use ; Heat-Shock Proteins ; metabolism ; physiology ; Humans ; Mutation ; Neurodegenerative Diseases ; drug therapy ; genetics ; metabolism ; pathology ; Parkinson Disease ; drug therapy ; genetics ; metabolism ; pathology ; Phenols ; isolation & purification ; therapeutic use ; Plants, Medicinal ; chemistry ; Proteasome Endopeptidase Complex ; metabolism ; Protein Folding ; Ubiquitin ; metabolism