摘要
背景
帕金森病(Parkinson's disease, PD)是最常见的神经系统退行性疾病之一,其病因与发病机制尚不明确,目前普遍认为可能与遗传、环境及年龄老化等多种因素相关,其发病率及患病率均随着年龄的增长而增加,65岁以上人群患病率约为1-2%,流行病学调查显示约10-15%的PD患者有阳性家族史。临床上以静止性震颤、运动迟缓、肌强直和姿势平衡障碍等为特征,主要病理改变为中脑黑质多巴胺能神经元选择性变性坏死和神经元胞浆中出现嗜酸性包涵体(Lewy小体)。研究发现约50%的黑质多巴胺能神经元丢失即可导致PD患者运动障碍症状的出现。
中脑多巴胺能神经元的正常发育涉及多种信号分子的共同调控,其中有两个转录因子在决定中脑多巴胺能神经元的形态、终末分化和存活发挥着重要作用,一个是孤儿核受体Nurrl,另一个是成对样同源结构域转录因子3(Paired-like homeodomain transcription factor-3, Pitx3)。国外有研究发现PITX3基因的单核苷酸多态性(single nucleotide polymorphism, SNP)可能增加罹患PD的危险性,目前国内尚无检测PITX3基因单核苷酸多态性与PD患者相关性的研究。
目的
探讨PITX3基因rs2281983、rs4919621、rs3758549三个单核苷酸多态性与中国人群散发性帕金森病(sporadic Parkinson's disease,SPD)的相关性。
方法
应用PCR-限制性片段长度多态性分析(Restriction fragment length polymorphism, RFLP)和DNA直接测序方法检测了512例中国散发性帕金森病患者和506例正常对照者PITX3基因rs2281983、rs4919621、rs3758549三个单核苷酸多态性,并进行多态性分析。
结果
对中国512例SPD患者(其中170例早发性PD,342例迟发性PD)和506例正常对照者检测了PITX3基因rs2281983、rs4919621、rs3758549三个单核苷酸多态性。SPD组rs2281983位点的基因型、等位基因频率分布与正常对照组比较差异无显著性(基因型：x2=0.217,P=0.897；等位基因频率：x2=0.128,P=0.721)。rs4919621位点在两组比较中同样无显著性差异(基因型：x2=0.949,P=0.622；等位基因频率：x2=0.318,P=0.573)。rs3758549位点的基因型、等位基因频率在SPD组与正常对照组比较差异有显著性(基因型：x2=7.868,P=0.020；等位基因频率：贮=5.475,P=0.019,OR=1.317,95%CI：1.045-1.660),且LOPD组与正常对照组比较同样有统计学意义(基因型：x2=9.843,P=0.007；等位基因频率：x2=4.603,P=0.032,OR=1.322,95%CI：1.024-1.707),EOPD组与正常对照组比较无显著差异性。
结论
PITX3基因的rs3758549多态可能是中国人群迟发性帕金森病的危险因素。不支持rs2281983,rs4919621多态为中国人群散发性帕金森病的危险因素。
Background
Parkinson's disease (PD) is one of the most common neurodegenerative diseases, the etiology and pathogenesis of which is unclear yet. It is widely accepted that genetic, environmental factors and progress of aging may be related to the mechanism of onset of PD. The incidence and prevalence rates increase with age and the prevalence of people over 65 years is about 1-2%. Epidemiological investigations revealed that about 10-15% of PD patients had positive family history. Clinically, PD is characterized by resting tremor, bradykinesia, rigidity, and postural instability. The pathology feature of it is the selective degeneration of the dopaminergic neurons in the substantia nigra and the formation of cytoplasmic eosinophilic inclusion bodies (Lewy body) in dopaminergic neurons in the brain. Study found that lossing about 50% of dopaminergic neurons can lead movement disorders in PD patients.
The normal development of midbrain dopaminergic neurons involves in co-regulation of multiple signaling molecules, including two transcription factors which play a crucial role in determining the dopaminergic neurons phenotype, terminal differentiation and survive. One is the orphan nuclear receptor Nurrl, the other is the paired-like homeodomain transcription factor Pitx3. Studies have found the single nucleotide polymorphism (SNP) of PITX3 gene may increase the risk for Parkinson's disease. There are still no research on Chinese people has been carried on.
Objective
Detect the prevalence of rs2281983, rs4919621, rs3758549 polymorphisms of PITX3 gene in Chinese patients with sporadic Parkinson's disease, to explore the association between them.
Method
We utilized PCR-Restricted Fragments Length Polymorphism (RFLP) and DNA sequencing to identify genotype of rs2281983, rs4919621, rs3758549 varients of PITX3 gene in 512 Chinese patients with SPD (including 170 cases of early onset PD,342 cases of late onset PD) and 506 unrelated healthy controls, then carried out an association analysis.
Result
There is no statistical difference in genotype or allele frequencies of rs2281983 variant in PITX3 gene between sporadic PD patient group and healthy control group (genotype:χ2=0.217, P=0.897; allele:χ2=0.128, P=0.721). It's the same of rs4919621 variant in PITX3 gene SPD patient group and healthy control group (genotype:χ2=0.949, P=0.622;allele:χ2=0.318, P=0.573). Rs3758549 variant in PITX3 gene get a statistical difference in genotype and allele frequencies between sporadic PD patient group and healthy control group (genotype:χ2=7.868, P=0.020; allele:χ2=5.475, P=0.019, OR=1.317,95%CI:1.045-1.660), and there is also a significant difference between LOPD patient group and healthy control group(genotype:χ2=9.843, P=0.007; allele:χ2=4.603, P=0.032, OR=1.322,95%CI:1.024-1.707). There is no statistical difference in genotype or allele frequencies when EOPD patient group compared with healthy control group.
Conclusion
1. Rs3758549 polymorphism in PITX3 gene is a risk for Chinese patients with LOPD.
2. Neither rs2281983 nor rs4919621 polymorphisms in PITX3 gene is a risk factor for the onset of PD in China.
引文
[1]Farrer MJ. Genetics of Parkinson's disease:paradigm shifts and future prospects. Nat Rev Genet,2006,7:306-318
[2]Sherer TB, Betarbet R, Greenamyre JT. Environment, mitochondria, and Parkinson's disease. Neuroscientist.2002,8:192-197
[3]Falin, S. Description of Parkinson's disease as a clinical syndrome. Ann N Y Acad Sci.2003,991,1-14
[4]Zhang ZX, Roman GC, Hong Z, et al. Parkinson's disease in China:Prevalence in Beijing, Xi'an and Shanghai. Lancet,2005.365:595-597
[5]Payami H, Larsen K, Bernard S, et al. Increased risk of Parkinson's disease in parents and siblings of patients. Ann Nenrol,1994,36(4):659-661
[6]Payami H, and Zareparsi, S. Genetic epidemiology of Parkinson's disease. Psychiatry Neurol.1998,11:98-106
[7]Leroy E, Boyer R, Auburger G, et al. The ubiquitin pathway in Parkinson's disease. Nature,1998,395(6701):451-452
[8]Paisan-Ruiz C, Jain S, Evans EW, et al. Cloning of the gene containing mutations that cause PARK8-linked Parkinson's disease. Neurol,2004,4(4): 595-600
[9]Lautier C, Goldwurm S, Durr A, et al. Mutations in the GIGYF2(TNRC 15)gene at the PARKll locus in familial Parkinson disease. Am J Hum Genet, 2008,82(4):822-33
[10]Kitada T, Asakawa S, Hattori N, et al. Mutations in the parkin gene cause automsomal recessive juvenile parkinsonism. Nature,1998,392(6676): 605-608
[11]Bonifati V, Rizzu P, vall Baren MJ, et al. Mutations in the DJ-1 gene associated with autosomal recessive early-onset Parkinsonism. Science,2003.299(5604): 256-259
[12]Valente EM, Abou-Sleiman PM, Caputo B, et al. Hereditary early-onset Parkinson's disease caused by mutations in PINKI. Science,2004,304(5674):1 158-1160
[13]Bonifati V. Genetics of Parkinson's disease. Hinerca Med,2005,96(3):175-86
[14]Strauss KM, Martins LM, Plun-Favreau H, et al. Loss of function mutations in the gene encoding Omi/HtrA2 in Parkinson's disease. Hum Mol Genet,2005, 14:2099-2111
[15]Coro Paisan-Ruiz, Kailash P, Bhatia, et al. Characterization of PLA2G6 as a Locus for Dystonia-Parkinsonism. Ann Neurol,2009,65:19-23
[16]A Di Fonzo, M.C.J. Dekker P, et al. FBX07 mutations cause autosomal recessive, early-onset parkinsonian-pyramidal syndrome. Neurology,2009,72: 127-136
[17]Singleton AB, Farter M, Johnson J, et al. Alpha-Synuclein locus triplication causes Parkinson's disease. Science,2003,302:841
[18]Chartier-Harlin MC, Kachergus J, Roumier C, ct al. Alpha-synuclein locus duplication as a cause of familial Parkinson's disease. Lancet,2004,364: 1167-1169
[19]Satake W, Nakabayashi Y, Mizuta I, et al. Genome-wide association study identifies common variants at four loci as genetic risk factors for Parkinson's disease. Neuro genetic,2009,41(12):1303-1307
[20]Fearnley JM, Lees AJ. Aging and parkinson's disease:substantia nigra regional electivity. Brain,1991,114:2283-2301
[21]Smidt MP, Smits SM, Peter JH, et al. Molecular mechanisms underlying midbrain dopamine neuron development and function. Eur J Pharmacol, 2003,480 (123):75-88
[22]Semina EV, Reiter RS, Murray JC, et al. Isolation of a new homeobox gene belonging to the Pitx/Rieg family:expression during lens development and mapping to the aphakia region on mouse chromosome 19. Hum Mol Genet, 1997,6:2109-2116
[23]Smidt MP, van Schaick HS, Lanctot C, et al. A homeodomain gene Ptx3 has highly restricted brain expression in mesencephalic dopaminergic neurons. Proc Natl Acad Sci,1997,94:13305-13310
[24]Vitalis T, Cases O, Engelkamp D, et al. Defect of tyrosine hydroxylase-immunoreacive neurons of mice lacking the transcription factor Pax6. Neurosci, 2000,20(17):6501-6516
[25]Varnum DS, Stevens LC. Aphakia, a new mutation in the mouse. Hered, 1968,59:147-150
[26]Semina EV, Murray JC, Reiter R, et, al. Deletion in the promoter region and altered expression of Pitx3 homeobox gene in aphakia mice. Hum Mol Genet, 2000,9(11):1575-1585
[27]Rieger DK, Reichenberger E, McLean W, et, al. A double-deletion mutation in the Pitx3 gene causes arrested lens development in aphakia mice. Genomics, 2001,72 (1):61-72
[28]Smidt MP, Smits SM, Bouwmeester H, et al. Early developmental failure of substantia nigra dopamine neurons in mice lacking the homeodomain gene Pitx3. Development,2004,131(5):1145-1155
[29]Nunes I, Tovamasian LT, Silva RM, et al. Pitx3 is required for development of substantia nigra dopaminergic neurons. Proe Natl Acad Sci,2003.100(7): 4245-4250
[30]Van Den Munckhof P, Luk KC, Ste-Marie L, et al. Pitx3 is required for motor activity and for survival of a subset of midbrain dopaminergic neurons. Development,2003,130(11):2535-2542
[31]Hwang DY, Fleming SM, Ardayfio P, et al.3,4-dihydroxy-phenylalanine reverses the motor deficits in Pitx3-deficient aphakin mice:behavioral characterization of a novel genetic model of Parkinson's disease. Neurosci, 2005,25(8):2132-2137
[32]Fuchs J, Mueller JC, Lichtner P, et al. The transcription factor PITX3 is associatedwith sporadic Parkinson's disease. Neurobiol Aging,2009,30(5): 731-738
[33]Bergman O, Hakansson A, Westberg L, et al.2008. PITX3 polymorphism is associated with early onset Parkinson's disease. Neurobiol Aging,2010,31(1): 114-117
[34]Haubenberger D, Reinthaler E, Mueller JC, et al. Association of transcription factor polymorphisms PITX3 and EN1 with Parkinson's disease. Neurobiol Aging.2009 Apr 2 [Epub ahead of print]
[35]Le W, Nguyen D, Lin XW, et al. Transcription factor PITX3 gene in Parkinson's disease. Neurobiol Aging.2009 Apr 24 [Epub ahead of print]
[36]Hughes AJ, Daniel SE, Kilford L, et al. Accuracy of clinical diagnosis of idiopathic Parkinson's disease:a clinico-pathological study of 100 cases. Neurol Neurosurg Psychiatry,1992,55(3):181-184
[37]Li Z, Zhang Z, He Z,et al. A partition-ligation-combination-subdivision EM algorithm for haplotype inference with multiallelic markers:update of the SHEsis. Cell Res,2009,19(4):519-23
[38]Ito G, Okai T, Fujino G, et al. GTP Binding is essential to the protein kinase activity of LRRK2, a causative gene product for familiar Parkinson's disease. Biochemistry,2007,46(5):1380-1388
[39]Semina EV, Reiter RS, Leysens NJ,et al. Cloning and characterization of a novel bicoid-related homeobox transcription factor gene, RIEG, involved in Rieger syndrome. Nature Genet,1996,14,392-399
[40]Zilinski CA, Shah R, Lane ME, et al. Modulaion of zebrafish pitx3 expression in the primordial of the pituitary, lens, olfactory epithelium and cranial ganglia by hedgehog and nodal signaling. Genesis,2005,41(1):33-40
[41]Shi X, Bosenko DV, Zinkevich NS, et al. Zebrafish pix3 is necessary for normal lens and retinal development. Mech. Dev,2005,122(4):513-527
[42]Jacobs FM, van der Linden AJ, Wang Y, et al. Identification of Dlkl, Ptpru and Klhll as novel Nurrl target genes in meso-diencephalic dopamine neurons. Development,2009,136(14):2363-73
[43]Cazorla P. Smidt, MP, O'Malley, KL, Burbach, JP. A response element for the homeodomain transcription factor Ptx3 in the tyrosine hydroxylase gene promoter. Neurochem,2000,74:1829-1837
[44]Lebel M, Gauthier Y, Moreau A, et al. Pitx3 activates mouse tyrosine hydroxylase promoter via a high-affinity binding site. Neurochem,2001,77: 558-567
[45]Smidt M, Asbreuk C, Cox J, et al. A second independent pathway for development of mesencephalic dopaminergic neurons requires Lmxlb. Nat Neurosci,2000,3(4):337-41
[46]Peng C, Fan S, Li X, Fan X et al. Overexpression of pitx3 upregulates expression of BDNF and GDNF in SH-SY5Y cells and primary ventral mesencephalic cultures. FEBS Le,2007,581(7):1357-61
[47]Yang D, Peng C, Li X, et al. Pitx3-transfected astrocytes secrete brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor and protect dopamine neurons in mesencephalon cultures. Neurosci Res,2008,86(15): 3393-3400
[48]Kim J, Inoue K, Ishii J, et al. A MicroRNA feedback circuit in midbrain dopamine neurons. Science,2007,317:1220-1224
[49]Cazorla P, Smidt MP, O'Malley KL, Burbach JP. A response element for the homeodomain transcription factor Ptx3 in the tyrosine hydroxylase gene promoter. Neurochem,2000,74:1829-1837
[50]Jacobs FM, Smits SM, Noorlander CW, et al. Retinoic acid counteracts developmental defects in the substantia nigra caused by Pitx3 deficiency. Development,2007,134:2673-2684
[1]Fahn S. Description of Parkinson's disease as a clinical syndrome. Ann NY Acad Sci,2003,991:-14
[2]Zhang ZX, Roman GC, Hong Z, et al. Parkinson's disease in China:Prevalence in Beijing, Xi'an and Shanghai. Lancet,2005,365:595-597
[3]Fearnley JM, Lees AJ. Ageing and Parkinson's disease:substantia nigra regional selectivity. Brain,1991,114 (5):2283-2301
[4]Lamonerie T, Tremblay JJ, Lanctot C, et al. Ptxl, a bicoid-related homeobox transcription factor involved in transcription of the pro-opiomelanocortin gene. Genes Dev,1996,10:1284-1295
[5]Gage PJ, Camper SA. Pituitary homeobox 2, a novel member of the bicoid-related family of homeobox genes, is a potential regulator of anterior structure formation. Hum Mol Genet,1997,6:457-464
[6]Semina EV, Reiter RS, Murray JC. Isolation of a new homeobox gene belonging to the Pitx/Rieg family:expression during lens development and mapping to the aphakia region on mouse chromosome 19. Hum Mol Genet,1997,6:2109-2116
[7]Smidt MP, van Schaick HS, Lanctot C, et al. A homeodomain gene Ptx3 has highly restricted brain expression in mesencephalic dopaminergic neurons. Proc Natl Acad Sci,1997,94:13305-13310
[8]Semina EV, Reiter RS, Leysens NJ,et al. Cloning and characterization of a novel bicoid-related homeobox transcription factor gene, RIEG, involved in Rieger syndrome. Nature Genet,1996,14,392-399
[9]Zilinski CA, Shah R, Lane ME, et al. Modulaion of zebrafish pitx3 expression in the primordial of the pituitary, lens, olfactory epithelium and cranial ganglia by hedgehog and nodal signaling. Genesis,2005,41(1):33-40
[10]Shi X, Bosenko DV, Zinkevich NS, et al. Zebrafish pix3 is necessary for normal lens and retinal development. Mech Dev,2005,122(4):513-527
[11]Varnum DS, Stevens LC. Aphakia, a new mutation in the mouse. Hered,1968,59: 147-150
[12]Semina EV, Murray J C, Reiter R, et al. Deletion in the promoter region and altered expression of Pitx3 homeobox gene in aphakia mice. Hum Mol Genet, 2000,9(11):1575-1585
[13]Rieger DK, Reichenberger E, McLean W, et al. A double-deletion mutation in the Pitx3 gene causes arrested lens development in aphakia mice. Genomics,2001, 72 (1):61-72
[14]Addison PKF, Berry V, Ionides ACW et al. Posterior polar cataract is the predominant consequence of a recurrent mutation in the PITX3 gene. Ophthalmol, 2005,89(2):138-141
[15]Coulon V, L'Honore A, Ouimette JF et al. A muscle-specific promoter directs pitx3 gene expression in skeletal muscle cells. Biol Chem,2007,282(45): 33192-33200
[16]Martinat C, Bacci JJ, Leete T, et al. Cooperative transcription activation by Nurrl and Pitx3 induces embryonic stem cell maturation to the midbrain dopamine neuron phenotype. Proc Natl Acad Sci,2006,103(8):2874-9
[17]Jacobs FM, van Erp S, van der Linden AJ, et al. Pitx3 potentiates Nurrl in dopamine neuron terminal differentiation through release of SMRT-mediated repression. Development,2009,136(4):531-40
[18]Jacobs FM, van der Linden AJ, Wang Y, et al. Identification of Dlkl, Ptpru and Klhll as novel Nurrl target genes in meso-diencephalic dopamine neurons. Development,2009,136(14):2363-73
[19]Smidt MP, Smits SM, Bouwmeester H, et al. Early developmental failure of substantia nigra dopamine neurons in mice lacking the homeodomain gene Pitx3. Development,2004,131(5):1145-1155
[20]Nunes I, Tovamasian LT, Silva RM, et al. Pitx3 is required for development of substantia nigra dopaminergic neurons. Proe Natl Acad Sci,2003,100(7): 4245-4250
[21]Van Den Munckhof P, Luk KC, Ste—Marie L, et al. Pitx3 is required for motor activity and for survival of a subset of midbrain dopaminergic neurons. Development,2003,130(11):2535-2542
[22]Hwang DY, Fleming SM, Ardayfio P, et al.3,4-dihydroxy-phenylalanine reverses the motor deficits in Pitx3-deficient aphakin mice:behavioral characterization of a novel genetic model of Parkinson's disease. Neurosci,2005,25(8):2132-2137
[23]Cazorla P. Smidt, MP, O'Malley, KL, Burbach, JP. A response element for the homeodomain transcription factor Ptx3 in the tyrosine hydroxylase gene promoter. Neurochem,2000,74:1829-1837
[24]Lebel M, Gauthier Y, Moreau A, et al. Pitx3 activates mouse tyrosine hydroxylase promoter via a high-affinity binding site. Neurochem,2001,77:558-567
[25]Smidt M, Asbreuk C, Cox J, et al. A second independent pathway for development of mesencephalic dopaminergic neurons requires Lmxlb. Nat Neurosci,2000,3(4):337-41
[26]Seroogy, KB, Lundgren KH, Tran TM, et al. Dopaminergic neurons in rat ventral midbrain express brain-derived neurotrophic factor and neurotrophin-3 mRNAs. Comp Neurol,1994,342:321-334
[27]Lin LF, Doherty DH, Lile JD, et al. GDNF:a glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons. Science,1993,260:1130-1132
[28]Peng C, Fan S, Li X, Fan X et al. Overexpression of pitx3 upregulates expression of BDNF and GDNF in SH-SY5Y cells and primary ventral mesencephalic cultures. FEBS Le,2007,581(7):1357-61
[29]Yang D, Peng C, Li X, et al. Pitx3-transfected astrocytes secrete brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor and protect dopamine neurons in mesencephalon cultures. Neurosci Res,2008,86(15): 3393-3400
[30]Jacobs FM, Smits SM, Noorlander CW, et al. Retinoic acid counteracts developmental defects in the substantia nigra caused by Pitx3 deficiency. Development,2007,134:2673-2684
[31]Kim J, Inoue K, Ishii J, et al. A MicroRNA feedback circuit in midbrain dopamine neurons. Science,2007,317:1220-1224
[32]Fuchs J, Mueller JC, Lichtner P, et al. The transcription factor PITX3 is associatedwith sporadic Parkinson's disease. Neurobiol Aging,2009,30(5): 731-738
[33]Bergman O, Hakansson A, Westberg L, et al.2008. PITX3 polymorphism is associated with early onset Parkinson's disease. Neurobiol Aging,2010,31(1): 114-117
[34]Haubenberger D, Reinthaler E, Mueller JC, et al. Association of transcription factor polymorphisms PITX3 and EN1 with Parkinson's disease. Neurobiol Aging.2009 Apr 2. [Epub ahead of print]
[35]Le W, Nguyen D, Lin XW, et al. Transcription factor PITX3 gene in Parkinson's disease. Neurobiol Aging.2009 Apr 24. [Epub ahead of print]
