DJ-1基因及其与帕金森病关系的研究进展

帕金森病是目前世界上最常见的神经系统退行性病变之一,DJ-1基因是一个与帕金森病相关的基因,其致病机制还不完全清楚.很多研究人员对DJ-1蛋白的功能进行了研究,目前已经发现DJ-1蛋白具有抗氧化、调节转录、参与能量代谢、抑制细胞凋亡等作用,本文总结了DJ-1基因的结构、突变形式、生物学作用及其与帕金森病发生的关系,以及DJ-1基因与其他导致帕金森病的基因之间的相互作用.对研究和了解帕金森病及其基因治疗具有重要意义.     

DJ-1在肿瘤中作用的研究进展

DJ-1基因是一种新的线粒体依赖癌基因。当前在帕金森病领域,对DJ-1基因的研究较多,认为其与人类家族性早发型帕金森病相关。近来研究发现DJ-1基因与人类多种恶性肿瘤的发生、进展及预后等密切相关,可作为癌症发生的预警标志物,但也有DJ-1在肿瘤中低表达的报道。     

DJ-1基因与帕金森氏病的研究进展

[摘要] 帕金森氏病是目前世界上最常见的神经系统退行性病变之一，DJ-1是一个与帕金森氏病相关的基因，其致病的作用机制不完全清楚。很多研究人员对DJ-1的蛋白功能进行了研究，目前已经发现DJ-1蛋白具有抗氧化、调节转录、参与能量代谢、抑制细胞凋亡等作用，本文总结了DJ-1的结构、突变形式、生物学作用与帕金森病发生的关系，以及DJ-1基因与其它导致帕金森氏病的基因之间的相互作用。对研究和了解帕金森氏病及其基因治疗具有重要意义。     

DJ-1在帕金森病和肿瘤中抗氧化作用的研究进展

氧化应激在帕金森病多巴胺能神经元变性中起着重要作用,活性氧(reactive oxygen species,ROS)产生的来源和机制包括多巴胺本身的代谢,线粒体功能障碍等。DJ-1在帕金森病和心力衰竭中起抗氧化作用,参与氧化应激保护细胞。DJ-1是癌症相关蛋白,它参与细胞内不同的信号通路。DJ-1可以调控氧化应激反应,对肿瘤细胞的侵袭转移行为产生了明确的影响,因此可以推测, DJ-1作为一种抗氧化剂可以影响肿瘤的治疗和复发。现在已证实DJ-1与癌症之间有着密切关系,但是它是如何作用的而改变细胞存活的信号通路的详细机制仍然未知。本文综述DJ-1在帕金森病,心衰,肿瘤中抗氧化机制,以及该靶点在肿瘤治疗中的研究进展。     

DJ-1抗氧化应激相关机制及其与帕金森发病的关系

帕金森病(Parkinson's disease,PD)是一种以黑质纹状体多巴胺能神经元选择性丢失和残存神经元胞质中出现嗜酸性包涵体即Lewy体为典型病理特征的进行性神经变性疾病。DJ-1基因突变后可引起常染色体隐性遗传的早发家族性PD。DJ-1蛋白的确切功能到目前为止还不清楚,有研究表明,DJ-1可通过自身氧化、调节抗氧化基因的转录,抑制ASK1活性,激活ERK1/2信号通路,阻止线粒体损伤所致的细胞凋亡。本文将主要对DJ-1抗氧化应激相关机制及其与帕金森病的关系简单综述。     

DJ-1蛋白在缺血性中风发病中的作用研究进展

人帕金森病蛋白7/DJ-1蛋白(PARK7/DJ-1)为帕金森病相关蛋白,近年来研究发现其可通过调节细胞内线粒体转位和自噬、利用细胞外旁分泌或自分泌方式保护缺血神经元、介导Nrf2/ARE通路上调谷胱甘肽、调节PTEN/PI3K/AKT信号通路调节缺血再灌注(IR)损伤,从而保护缺血后脑组织,有望成为缺血性卒中新的生物标志物及治疗靶点。本文从DJ-1蛋白概况、DJ-1蛋白在缺血性中风发病中的作用等方面对近年来DJ-1蛋白在缺血性中风发病中的研究进展进行归纳总结,并分析其作为缺血性卒中治疗靶点的优势及目前研究中存在的不足。     

Parkinson病遗传学研究进展

帕金森病是最常见的伴有运动障碍的神经变性病,虽然该病多为散发病例,但至少有10个以上的位点和它相关。帕金森病的遗传学研究为理解它的发病机制提供重要线索。新近又发现PINK1的W4370DA突变和DJ-1基因的敲除及T497C突变是遗传性帕金森病的病因。     

Parkinson病遗传学研究进展

帕金森病是最常见的伴有运动障碍的神经变性病，虽然该病多为散发病例，但至少有10个以上的位点和它相关。帕金森病的遗传学研究为理解它的发病机制提供重要线索。新近又发现PINK1的W4370DA突变和DJ-1基因的敲除及T497C突变是遗传性帕金森病的病因。     

DJ-1缺失增加出生前脂多糖暴露引起的出生后小鼠多巴胺能神经元丢失(英文)

目的:观察DJ-1缺失和出生前脂多糖(lipopolysaccharide,LPS)暴露对出生后小鼠多巴胺能(dopam-inergic,DA)系统和神经炎症的影响。方法:妊娠第10.5 d DJ-1基因敲除的小鼠腹腔注射脂多糖(10,000 EU/kg体重)后自然分娩的后代在4月和14月龄时处死。免疫组织化学染色结合体视学计数定量黑质酪氨酸羟化酶(Tyrosine hydroxylase,TH)和CD11b阳性细胞数量。高效液相色谱法测定纹状体DA代谢物水平。免疫荧光法测定TNF-α和IL-1β蛋白水平。结果:4月和14月龄的DJ-1基因敲除的小鼠均没有明显的DA神经元减少和脑内神经炎症改变。出生前接触过LPS的4月和14月龄野生型小鼠,黑质DA能神经元分别减少13.6%和23.1%,纹状体DA含量分别减少19.0%和26.2%,同时神经炎症改变明显。出生前接触过LPS的4月和14月龄DJ-1基因敲除小鼠,黑质DA能神经元分别减少22.5%和35.4%,纹状体DA含量分别减少34.3%和39.3%,同时脑内炎症反应更明显。结论:以上结果提示遗传因素缺失和环境因素可能共同引发帕金森病发生,进一步验证了帕金森病多因素损伤引发的发病假说。     

MicroRNA在建立DJ-1基因敲减细胞模型中的应用

 目的 与传统的siRNA方法建立基因敲减模型不同,探讨合成microRNA(miRNA)在DJ-1基因敲减细胞模型中的应用。方法 构建针对DJ-1基因的合成miRNA载体,合成DJ-1基因的siRNA干扰片断。在脂质体介导下,将上述两种小干扰RNA载体或片断分别转染MN9D细胞系,应用real-time PCR和Western blots检测目的基因DJ-1的mRNA和蛋白表达。结果 与Control组相比,转染合成microRNA的MN9D细胞中,DJ-1的mRNA水平下调90%（p<0.05）,蛋白水平下调70%~85%(p<0.05);而转染siRNA的MN9D细胞中,DJ-1的mRNA水平下调50%~70%（p<0.05）,蛋白水平下调20%~50%（p<0.05）。结论 microRNA与siRNA均可作为基因敲减的重要研究手段。与传统的siRNA相比,合成microRNA对目的DJ-1的干扰效率更为显著。     

Detection of Parkin (PARK2) and DJ1 (PARK7) mutations in early-onset Parkinson disease: Parkin mutation frequency depends on ethnic origin of patients

Mutations in the Parkin (PARK2) and the DJ1 (PARK7) gene cause early-onset Parkinson disease (EOPD). We tested 75 Serbian EOPD patients for mutations in both genes by conventional mutational screening (SSCP/dHPLC/sequencing) to detect small sequence alterations and by gene dosage studies (quantitative PCR) to reveal deletions or multiplications of one or more exons. A compound heterozygous Parkin mutation (exon deletion and point mutation; [c.836_972del]+[c.1411C>T]; +1 is first nucleotide of GenBank AB009973.1) was identified in a patient who showed a relatively benign course after a disease onset at 41 years. Another case had a heterozygous exon deletion in DJ1 ([c.253_322del]+[?]) and presented with an age at onset of 45 years and a rapid disease course. In conclusion, Parkin mutations are surprisingly rare in our Serbian EOPD sample, suggesting that the mutation rate depends on the ethnic origin of the patients. Although DJ1 mutations appear to be rare, we confirm their role in EOPD and demonstrate the importance of gene dosage studies.     

Novel homozygous p.E64D mutation in DJ1 in early onset Parkinson disease (PARK7)

Mutations in the parkin gene have been identified as a common cause of autosomal recessive inherited Parkinson disease (PD) associated with early disease manifestation. However, based on linkage data, mutations in other genes contribute to the genetic heterogeneity of early-onset PD (EOPD). Recently, two mutations in the DJ1 gene were described as a second cause of autosomal recessive EOPD (PARK7). Analyzing the PARK7/DJ1 gene in 104 EOPD patients, we identified a third mutation, c.192G>C (p.E64D), associated with EOPD in a patient of Turkish ancestry and characterized the functional significance of this amino acid substitution. In the patient, a substantial reduction of dopamine uptake transporter (DAT) binding was found in the striatum using [(18)F]FP-CIT and PET, indicating a serious loss of presynaptic dopaminergic afferents. His sister, homozygous for E64D, was clinically unaffected but showed reduced dopamine uptake when compared with a clinically unaffected brother, who is heterozygous for E64D. We demonstrate by crystallography that the E64D mutation does not alter the structure of the DJ1 protein, however we observe a tendency towards decreased levels of the mutant protein when overexpressed in HEK293 or COS7 cells. Using immunocytochemistry in contrast to the homogenous nuclear and cytoplasmic staining in HEK293 cells overexpressing wild-type DJ1, about 5% of the cells expressing E64D and up to 80% of the cells expressing the recently described L166P mutation displayed a predominant nuclear localization of the mutant DJ1 protein.     

Pathogenic mutations in Parkinson disease

Parkinson disease (PD; Parkinson's) is the second most common neurodegenerative disease, characterized by the progressive loss of dopamine neurons and the accumulation of Lewy bodies. Increasing evidence suggests that deficits in mitochondrial function, oxidative and nitrosative stress, the accumulation of aberrant or misfolded proteins, and ubiquitin-proteasome system (UPS) dysfunction may represent the principal molecular pathways that commonly underlie the pathogenesis. The relative role of genetic and environmental factors has been the focus of research and debate. The recent discovery of a number of disease-causing genes (SNCA, Parkin/PARK2, UCHL1, PINK1, DJ1/PARK7, and LRRK2) in familial and sporadic forms of PD has provided considerable insights into the pathophysiology of this complex disorder. The frequency of these gene mutations may vary according to ethnicity and to the specific gene. A gene dosage effect is observed in some cases, and the phenotype of some of the mutation carriers closely resembles typical PD. Penetrance of some of the recurrent mutations is incomplete and may vary with age. Further research to unravel the etiopathology could identify biochemical or genetic markers for potential neuroprotective trials.     

Unexpected mitochondrial matrix localization of Parkinson's disease‐related DJ‐1 mutants but not wild‐type DJ‐1

DJ‐1 has been identified as a gene responsible for recessive familial Parkinson's disease (familial Parkinsonism), which is caused by a mutation in the PARK7 locus. Consistent with the inferred correlation between Parkinson's disease and mitochondrial impairment, mitochondrial localization of DJ‐1 and its implied role in mitochondrial quality control have been reported. However, the mechanism by which DJ‐1 affects mitochondrial function remains poorly defined, and the mitochondrial localization of DJ‐1 is still controversial. Here, we show the mitochondrial matrix localization of various pathogenic and artificial DJ‐1 mutants by multiple independent experimental approaches including cellular fractionation, proteinase K protection assays, and specific immunocytochemistry. Localization of various DJ‐1 mutants to the matrix is dependent on the membrane potential and translocase activity in both the outer and the inner membranes. Nevertheless, DJ‐1 possesses neither an amino‐terminal alpha‐helix nor a predictable matrix‐targeting signal, and a post‐translocation processing‐derived molecular weight change is not observed. In fact, wild‐type DJ‐1 does not show any evidence of mitochondrial localization at all. Such a mode of matrix localization of DJ‐1 is difficult to explain by conventional mechanisms and implies a unique matrix import mechanism for DJ‐1 mutants.     

家族性帕金森病α-共核蛋白基因第3、4外显子的初步研究

目的 分析家族性帕金森病与α-共核蛋白基因的相关性，在其第3、4外显子中寻找相关突变或多态。方法 收集中国帕金森病家系，采用单链构象多态性(single strand conformational polymorphism，SSCP)与异源双链(heteroduplex analysis，HA)分析相结合的方法，筛查α-共核蛋白第3、4外显子中是否存在致病性突变。结果 SSCP和HA分析第3、4外显子未见单双链泳动异常。基因测序发现第4内含子5′端的第23位和67位分别插入1个c和t。结论 (1)α-共核蛋白基因的第3、4外显子不是中国家族性帕金森病的突变热点；(2)发现α-共核蛋白基因第4内含子在不同人群中有两个多态位点(23ins c和67ins t)。     

DJ‐1 is a Parkinson's disease susceptibility gene in southern Italy

De Marco EV, Annesi G, Tarantino P, Nicoletti G, Civitelli D, Messina D, Annesi F, Arabia G, Salsone M, Condino F, Novellino F, Provenzano G, Rocca FE, Colica C, Morelli M, Scornaienchi V, Greco V, Giofrè L, Quattrone A. DJ‐1 is a Parkinson's disease susceptibility gene in southern Italy. Mutations in the gene DJ‐1 have been shown to be a rare cause of early‐onset Parkinson's disease (EOPD). Since DJ‐1 mutations have been found in patients with Parkinson's disease (PD) from southern Italy, we aimed to investigate whether polymorphisms within the DJ‐1 gene could represent a risk factor for sporadic PD. First, we genotyped 294 patients with PD and 298 controls coming from southern Italy to assess the distribution of the insertion/deletion (Ins/Del) polymorphism. In a second phase, we identified five single‐nucleotide polymorphisms (SNPs) useful to delimit a region potentially involved and genotyped all patients and controls for these markers. All the markers analyzed were significantly associated with PD at both allelic and genotypic level. The most significant association with the disease was found at the Ins/Del polymorphism (p = 0.0001; adjusted odds ratio (OR ) = 2.05; confidence interval (CI ) = 1.36–3.08). When we considered a three‐marker sliding window, we found a highly significant association between the disease and the haplotypes including markers rs17523802, Ins/Del, and rs3766606 (p = 0.0007) and markers Ins/Del, rs3766606 and rs7517357 (p = 0.0054). Our results indicate that polymorphisms located in a region spanning 3535 bp from the promoter to the intron 2 of the DJ‐1 gene confer risk to sporadic PD in southern Italy.     

Measurement of recombinase activity in a set of related Abelson murine leukemic virus pre-B cell lines: DJ/DJ lines have more recombinase activity than do VDJ/VDJ lines

The VDJ recombination potential of a number of Abelson murine leukemic virus transformed fetal liver cell lines derived from (C57BL/6 x BALB/c) F1 mice was measured. The specific developmental stage of each line was determined using Southern blot analysis to ascertain their rearrangement status at the immunoglobulin heavy chain locus (DJ/DJ, VDJ/DJ or VDJ/VDJ). It was observed that DNA from DJ/DJ lines gave many more 'subhaploid' bands hybridizing with JH than did DNA from VDJ/VDJ lines. While the lack of appropriate substrate (VDJ/VDJ lines have exhausted the normal IgH substrate) contributes to the decrease in 'subhaploid bands', this result indicates that the rate of ongoing immunoglobulin heavy chain gene rearrangement was higher in the DJ/DJ lines. However, when the lines were examined using the assay developed by Hesse et al. (4) to measure VDJ recombinase activity, it was found that although all lines had recombinase activity, the DJ/DJ lines had four times more VDJ recombinase activity than did the VDJ/VDJ lines.     

Genetic etiology of Parkinson disease associated with mutations in the SNCA,PARK2, PINK1, PARK7, and LRRK2 genes: a mutation update

To date, molecular genetic analyses have identified over 500 distinct DNA variants in five disease genes associated with familial Parkinson disease; α‐synuclein (SNCA), parkin (PARK2), PTEN‐induced putative kinase 1 (PINK1), DJ‐1 (PARK7), and Leucine‐rich repeat kinase 2 (LRRK2). These genetic variants include ～82% simple mutations and ～18% copy number variations. Some mutation subtypes are likely underestimated because only few studies reported extensive mutation analyses of all five genes, by both exonic sequencing and dosage analyses. Here we present an update of all mutations published to date in the literature, systematically organized in a novel mutation database ( http://www.molgen.ua.ac.be/PDmutDB ). In addition, we address the biological relevance of putative pathogenic mutations. This review emphasizes the need for comprehensive genetic screening of Parkinson patients followed by an insightful study of the functional relevance of observed genetic variants. Moreover, while capturing existing data from the literature it became apparent that several of the five Parkinson genes were also contributing to the genetic etiology of other Lewy Body Diseases and Parkinson‐plus syndromes, indicating that mutation screening is recommendable in these patient groups. Hum Mutat 31:763–780, 2010. © 2010 Wiley‐Liss, Inc.     

常染色体隐性遗传性早发型帕金森综合征DJ1基因突变研究

目的探讨常染色体隐性遗传性早发型帕金森综合征（autosomal recessive early-onset Parkinsonism，AR—EP）DJ1基因的突变特点。方法 应用聚合酶链反应结合DNA直接序列分析方法，对11个常染色体隐性遗传性早发型帕金森综合征家系先证者的DJ1基因进行突变研究。结果本组AR-EP患者未发现DJ1基因的致病突变，在内含子区发现6个多态，分别为IVS1→15T→C、IVS4＋30T→G、TVS4＋45G→A、IVS4＋46G→A、IVS5＋31G→A和g．168-185del，其中3个（IVS1-15→C、IVS4＋45G→A、IVS4＋46G→A）为新发现的多念。结论中国人常染色体隐性遗传性早发型帕金森综合征患者DJ1基因突变可能罕见。     

Complete screening for glucocerebrosidase mutations in Parkinson disease patients from Portugal

Mutations in the gene encoding beta-glucocerebrosidase, a lysosomal degrading enzyme, have recently been associated with the development of Parkinson disease.Here we report the results found in a cohort of Portuguese Parkinson disease patients and healthy age-matched controls for mutations in the aforementioned gene. This screening was accomplished by sequencing the complete open-reading frame, as well as intron/exon boundaries, of the glucocerebrosidase gene, in a total of 230 patients and 430 controls.We have found an increased number of Parkinson disease patients presenting mutations in GBA when compared to controls.These results, together with recent literature, clearly suggest a role of glucocerebrosidase in the development of Parkinson disease.