脆X相关失调症检测方法的研究进展
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摘要
脆性X相关失调症是一类由FMR1基因突变引起的失调症,包括脆X相关震颤/共济失调综合征(FXTAS)、脆性X相关原发性卵巢机能不全(FXPOI)、脆X综合征等。这些失调症绝大多数由FMR1基因5′末端(CGG)n重复片段异常扩增和异常甲基化导致的FMRP蛋白合成减少或缺失所致,尚无明确的临床诊断标准,一般依赖于医师个人评估,本文将对Southern blotting,PCR,蛋白检测等方法做出介绍,以期为临床医师提供参考。
Fragile X-related disorders are a group of disorders caused by mutations in the FMR1 gene,including fragile X-associated tremor/ataxia syndrome(FXTAS),fragile X-associated primary ovarian insufficiency(FXPOI),and fragile X syndrome. Most of these disorders are caused by instability of a CGG-repeat tract at the 5′ end of the FMR1 transcript or abnormal methylation of the CpG residues in fanking region of FMR1 gene,these mutations will lead to the expression of FMPR reduced or silenced. There is no clear clinical diagnostic criteria yet,this article will introduce the detection methods as Southern blotting,PCR,protein detection,in order to provide reference for clinicians.
Fragile X-related disorders are a group of disorders caused by mutations in the FMR1 gene,including fragile X-associated tremor/ataxia syndrome(FXTAS),fragile X-associated primary ovarian insufficiency(FXPOI),and fragile X syndrome. Most of these disorders are caused by instability of a CGG-repeat tract at the 5′ end of the FMR1 transcript or abnormal methylation of the CpG residues in fanking region of FMR1 gene,these mutations will lead to the expression of FMPR reduced or silenced. There is no clear clinical diagnostic criteria yet,this article will introduce the detection methods as Southern blotting,PCR,protein detection,in order to provide reference for clinicians.
引文
[1]詹建英.脆X综合征的分子基础及其认知研究进展[J].国外医学儿科学分册,2005,32(2):111-113.
[2]Hayward BE,Kumari D,Usdin K.Recent advances in assays for the fragile X-related disorders[J].Hum Genet,2017,136(10):1313-1327.doi:10.1007/s00439-017-1840-5.
[3]邬玲仟潘乾,龙志高,等.脆性X综合征的基因诊断与产前诊断[J].遗传,2003,25(2):123-128.
[4]Monaghan KG,Lyon E,Spector EB.ACMG Standards and Guidelines for fragile X testing:a revision to the disease-specific supplements to the Standards and Guidelines for Clinical Genetics Laboratories of the American College of Medical Genetics and Genomics[J].Genet Med,2013,15(7):575-586.doi:10.1038/gim.2013.61.
[5]L.SS.Premature ovarian failure in the fragile X syndrome[J].American Journal of Medical Genetics,2000,97(3):189-194.doi:doi:10.1002/1096-8628(200023)97:3<189::AID-AJMG1036>3.0.CO;2-J.
[6]Tassone F,Adams J,Berry-Kravis EM,et al.CGG repeat length correlates with age of onset of motor signs of the fragile X-associated tremor/ataxia syndrome(FXTAS)[J].Am J Med Genet B Neuropsychiatr Genet,2007,144B(4):566-569.doi:10.1002/ajmg.b.30482.
[7]Loesch D,Hagerman R.Unstable Mutations in the FMR1 Gene and the Phenotypes[A].In:Tandem Repeat Polymorphisms:Genetic Plasticity,Neural Diversity and Disease(Hannan AJ,ed).New York,NY:Springer New York,2012:78-114.
[8]Latham GJ,Coppinger J,Hadd AG,et al.The role of AGGinterruptions in fragile X repeat expansions:a twenty-year perspective[J].Front Genet,2014,5:244.doi:10.3389/fgene.2014.00244.
[9]Nolin SL,Glicksman A,Ersalesi N,et al.Fragile X full mutation expansions are inhibited by one or more AGG interruptions in premutation carriers[J].Genet Med,2015,17(5):358-364.doi:10.1038/gim.2014.106.
[10]Yrigollen CM,Martorell L,Durbin-Johnson B,et al.AGGinterruptions and maternal age affect FMR1 CGG repeat allele stability during transmission[J].J Neurodev Disord,2014,6(1):24.doi:10.1186/1866-1955-6-24.
[11]Oh SY,He F,Krans A,et al.RAN translation at CGG repeats induces ubiquitin proteasome system impairment in models of fragile X-associated tremor ataxia syndrome[J].Hum Mol Genet,2015,24(15):4317-4326.doi:10.1093/hmg/ddv165.
[12]Sellier C,Freyermuth F,Tabet R,et al.Sequestration of DROSHAand DGCR8 by expanded CGG RNA repeats alters microRNAprocessing in fragile X-associated tremor/ataxia syndrome[J].Cell Rep,2013,3(3):869-880.doi:10.1016/j.celrep.2013.02.004.
[13]花茂方,刘小云,王文,等.脆性X综合征FMR1基因CGG重复序列与甲基化的检测[J].中国优生与遗传杂志,2012,20(06):18-20.
[14]Gold B,Radu D,Balanko A,et al.Diagnosis of Fragile Xsyndrome by Southern blot hybridization using a chemiluminescent probe:a laboratory protocol[J].Mol Diagn,2000,5(3):169-178.doi:10.1054/modi.2000.9404.
[15]Yu A,Barron MD,Romero RM,et al.At physiological pH,d(CCG)15 forms a hairpin containing protonated cytosines and a distorted helix[J].Biochemistry,1997,36(12):3687-3699.doi:10.1021/bi9625410.
[16]Filipovic-Sadic S,Sah S,Chen L,et al.A novel FMR1 PCRmethod for the routine detection of low abundance expanded alleles and full mutations in fragile X syndrome[J].Clin Chem,2010,56(3):399-408.doi:10.1373/clinchem.2009.136101.
[17]Hayward BE,Zhou Y,Kumari D,et al.A Set of Assays for the Comprehensive Analysis of FMR1 Alleles in the Fragile X-Related Disorders[J].J Mol Diagn,2016,18(5):762-774.doi:10.1016/j.jmoldx.2016.06.001.
[18]Chen L,Hadd A,Sah S,et al.High-resolution methylation polymerase chain reaction for fragile X analysis:evidence for novel FMR1 methylation patterns undetected in Southern blot analyses[J].Genet Med,2011,13(6):528-538.doi:10.1097/GIM.0b013e31820a780f.
[19]Groth KA,Skakkebaek A,Host C,et al.Clinical review:Klinefelter syndrome--a clinical update[J].J Clin Endocrinol Metab,2013,98(1):20-30.doi:10.1210/jc.2012-2382.
[20]Tassone F,Beilina A,Carosi C,et al.Elevated FMR1 mRNA in premutation carriers is due to increased transcription[J].RNA,2007,13(4):555-562.doi:10.1261/rna.280807.
[21]Hayward BE,Usdin K.Improved Assays for AGG Interruptions in Fragile X Premutation Carriers[J].J Mol Diagn,2017,19(6):828-835.doi:10.1016/j.jmoldx.2017.06.008.
[22]柳延虎,王璐,于黎.单分子实时测序技术的原理与应用[J].遗传,2015,37(03):259-268.
[23]乌日拉嘎,徐海燕,冯淑贞,等.测序技术的研究进展及三代测序的应用[J].中国乳品工业,2016,44(04):33-37.
[24]Kaufmann M,Schuffenhauer A,Fruh I,et al.High-Throughput Screening Using iPSC-Derived Neuronal Progenitors to Identify Compounds Counteracting Epigenetic Gene Silencing in Fragile X Syndrome[J].J Biomol Screen,2015,20(9):1101-1111.doi:10.1177/1087057115588287.
[25]Iwahashi C,Tassone F,Hagerman RJ,et al.A quantitative ELISAassay for the fragile x mental retardation 1 protein[J].J Mol Diagn,2009,11(4):281-289.doi:10.2353/jmoldx.2009.080118.
[26]LaFauci G,Adayev T,Kascsak R,et al.Fragile X screening by quantification of FMRP in dried blood spots by a Luminex immunoassay[J].J Mol Diagn,2013,15(4):508-517.doi:10.1016/j.jmoldx.2013.02.006.
[27]D GA.A fragile X case with an amplification/deletion mosaic pattern[J].Hum Genet,2000,3(106).
[28]邬玲仟,张学,等.医学遗传学[M].北京:人民卫生出版社,2016.
[2]Hayward BE,Kumari D,Usdin K.Recent advances in assays for the fragile X-related disorders[J].Hum Genet,2017,136(10):1313-1327.doi:10.1007/s00439-017-1840-5.
[3]邬玲仟潘乾,龙志高,等.脆性X综合征的基因诊断与产前诊断[J].遗传,2003,25(2):123-128.
[4]Monaghan KG,Lyon E,Spector EB.ACMG Standards and Guidelines for fragile X testing:a revision to the disease-specific supplements to the Standards and Guidelines for Clinical Genetics Laboratories of the American College of Medical Genetics and Genomics[J].Genet Med,2013,15(7):575-586.doi:10.1038/gim.2013.61.
[5]L.SS.Premature ovarian failure in the fragile X syndrome[J].American Journal of Medical Genetics,2000,97(3):189-194.doi:doi:10.1002/1096-8628(200023)97:3<189::AID-AJMG1036>3.0.CO;2-J.
[6]Tassone F,Adams J,Berry-Kravis EM,et al.CGG repeat length correlates with age of onset of motor signs of the fragile X-associated tremor/ataxia syndrome(FXTAS)[J].Am J Med Genet B Neuropsychiatr Genet,2007,144B(4):566-569.doi:10.1002/ajmg.b.30482.
[7]Loesch D,Hagerman R.Unstable Mutations in the FMR1 Gene and the Phenotypes[A].In:Tandem Repeat Polymorphisms:Genetic Plasticity,Neural Diversity and Disease(Hannan AJ,ed).New York,NY:Springer New York,2012:78-114.
[8]Latham GJ,Coppinger J,Hadd AG,et al.The role of AGGinterruptions in fragile X repeat expansions:a twenty-year perspective[J].Front Genet,2014,5:244.doi:10.3389/fgene.2014.00244.
[9]Nolin SL,Glicksman A,Ersalesi N,et al.Fragile X full mutation expansions are inhibited by one or more AGG interruptions in premutation carriers[J].Genet Med,2015,17(5):358-364.doi:10.1038/gim.2014.106.
[10]Yrigollen CM,Martorell L,Durbin-Johnson B,et al.AGGinterruptions and maternal age affect FMR1 CGG repeat allele stability during transmission[J].J Neurodev Disord,2014,6(1):24.doi:10.1186/1866-1955-6-24.
[11]Oh SY,He F,Krans A,et al.RAN translation at CGG repeats induces ubiquitin proteasome system impairment in models of fragile X-associated tremor ataxia syndrome[J].Hum Mol Genet,2015,24(15):4317-4326.doi:10.1093/hmg/ddv165.
[12]Sellier C,Freyermuth F,Tabet R,et al.Sequestration of DROSHAand DGCR8 by expanded CGG RNA repeats alters microRNAprocessing in fragile X-associated tremor/ataxia syndrome[J].Cell Rep,2013,3(3):869-880.doi:10.1016/j.celrep.2013.02.004.
[13]花茂方,刘小云,王文,等.脆性X综合征FMR1基因CGG重复序列与甲基化的检测[J].中国优生与遗传杂志,2012,20(06):18-20.
[14]Gold B,Radu D,Balanko A,et al.Diagnosis of Fragile Xsyndrome by Southern blot hybridization using a chemiluminescent probe:a laboratory protocol[J].Mol Diagn,2000,5(3):169-178.doi:10.1054/modi.2000.9404.
[15]Yu A,Barron MD,Romero RM,et al.At physiological pH,d(CCG)15 forms a hairpin containing protonated cytosines and a distorted helix[J].Biochemistry,1997,36(12):3687-3699.doi:10.1021/bi9625410.
[16]Filipovic-Sadic S,Sah S,Chen L,et al.A novel FMR1 PCRmethod for the routine detection of low abundance expanded alleles and full mutations in fragile X syndrome[J].Clin Chem,2010,56(3):399-408.doi:10.1373/clinchem.2009.136101.
[17]Hayward BE,Zhou Y,Kumari D,et al.A Set of Assays for the Comprehensive Analysis of FMR1 Alleles in the Fragile X-Related Disorders[J].J Mol Diagn,2016,18(5):762-774.doi:10.1016/j.jmoldx.2016.06.001.
[18]Chen L,Hadd A,Sah S,et al.High-resolution methylation polymerase chain reaction for fragile X analysis:evidence for novel FMR1 methylation patterns undetected in Southern blot analyses[J].Genet Med,2011,13(6):528-538.doi:10.1097/GIM.0b013e31820a780f.
[19]Groth KA,Skakkebaek A,Host C,et al.Clinical review:Klinefelter syndrome--a clinical update[J].J Clin Endocrinol Metab,2013,98(1):20-30.doi:10.1210/jc.2012-2382.
[20]Tassone F,Beilina A,Carosi C,et al.Elevated FMR1 mRNA in premutation carriers is due to increased transcription[J].RNA,2007,13(4):555-562.doi:10.1261/rna.280807.
[21]Hayward BE,Usdin K.Improved Assays for AGG Interruptions in Fragile X Premutation Carriers[J].J Mol Diagn,2017,19(6):828-835.doi:10.1016/j.jmoldx.2017.06.008.
[22]柳延虎,王璐,于黎.单分子实时测序技术的原理与应用[J].遗传,2015,37(03):259-268.
[23]乌日拉嘎,徐海燕,冯淑贞,等.测序技术的研究进展及三代测序的应用[J].中国乳品工业,2016,44(04):33-37.
[24]Kaufmann M,Schuffenhauer A,Fruh I,et al.High-Throughput Screening Using iPSC-Derived Neuronal Progenitors to Identify Compounds Counteracting Epigenetic Gene Silencing in Fragile X Syndrome[J].J Biomol Screen,2015,20(9):1101-1111.doi:10.1177/1087057115588287.
[25]Iwahashi C,Tassone F,Hagerman RJ,et al.A quantitative ELISAassay for the fragile x mental retardation 1 protein[J].J Mol Diagn,2009,11(4):281-289.doi:10.2353/jmoldx.2009.080118.
[26]LaFauci G,Adayev T,Kascsak R,et al.Fragile X screening by quantification of FMRP in dried blood spots by a Luminex immunoassay[J].J Mol Diagn,2013,15(4):508-517.doi:10.1016/j.jmoldx.2013.02.006.
[27]D GA.A fragile X case with an amplification/deletion mosaic pattern[J].Hum Genet,2000,3(106).
[28]邬玲仟,张学,等.医学遗传学[M].北京:人民卫生出版社,2016.