鄭清文 詹國(guó)棟 唐 春 畢 欣 趙薇薇 鄒小兵

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·論著·

CHD8基因新發(fā)突變的孤獨(dú)癥譜系障礙1例報(bào)道并文獻(xiàn)復(fù)習(xí)

鄭清文1詹國(guó)棟1唐 春1畢 欣2趙薇薇2鄒小兵1

目的 提高對(duì)孤獨(dú)癥譜系障礙(ASD)CHD8基因突變患兒臨床表型和基因型的認(rèn)識(shí)。方法回顧性分析1例存在CHD8基因突變的ASD患兒的臨床資料，并文獻(xiàn)復(fù)習(xí)。結(jié)果①男，3歲3月齡，頭圍55.0 cm(>4s)，身高107.0 cm(>2s)，體重23.5 kg(>2s)。有典型的ASD表現(xiàn)，運(yùn)動(dòng)明顯落后，有慢性便秘史。②提取患兒及其父母靜脈血DNA，以二代高通量測(cè)序技術(shù)對(duì)4 813個(gè)臨床相關(guān)基因的外顯子區(qū)進(jìn)行測(cè)序，共檢測(cè)到變異8 982個(gè)，經(jīng)篩選流程篩選ASD相關(guān)的EHMT1、PCDH9、NLGN4X的錯(cuò)義突變，CHD8的無(wú)義突變(c.307C>T, p.Gln103*)有致病可能。Sanger測(cè)序顯示患兒EHMT1、PCDH9的突變來(lái)源于父親，NLGN4X的突變來(lái)源于母親，患兒父母未發(fā)現(xiàn)CHD8的突變。③目前國(guó)外共報(bào)道了CHD8基因與神經(jīng)發(fā)育障礙有關(guān)的15個(gè)重要突變；CHD8基因突變患者的臨床表型包括ASD表現(xiàn)(87%)、身材高大(86%)、大頭畸形(80%)、慢性便秘或間歇性腹瀉(80%)、運(yùn)動(dòng)落后(67%)和睡眠異常(67%)等，本文病例臨床表型與文獻(xiàn)報(bào)道較為一致。結(jié)論CHD8基因是ASD重要的易感基因；ASD患兒合并大頭畸形、生長(zhǎng)過(guò)快，且有慢性便秘或間歇性腹瀉時(shí)應(yīng)考慮CHD8基因突變可能，可行基因檢測(cè)協(xié)助診斷。

孤獨(dú)癥譜系障礙；CHD8基因； 大頭畸形； 生長(zhǎng)過(guò)快； 二代測(cè)序技術(shù)

1 病例資料

母親孕早期出現(xiàn)陰道流血，服中藥及臥床休息后流血停止；定期產(chǎn)檢，未見(jiàn)異常，無(wú)妊娠期發(fā)熱，否認(rèn)貓、狗、放射線等不良接觸史。否認(rèn)家族中有神經(jīng)精神發(fā)育異常者。

查體：身高107.0 cm(>2s)，體重23.5 kg(>2s)，頭圍55.0 cm(>4s)。前額寬且突出。心音有力，律齊，未聞及病理性雜音；腹軟，未捫及包塊，未見(jiàn)臍疝、腹股溝疝；脊柱、指/趾端未見(jiàn)畸形；手掌皮紋未見(jiàn)異常。

實(shí)驗(yàn)室檢查：肝功能、血脂、甲狀腺功能、血漿皮質(zhì)醇(上午8時(shí))、胰島素樣生長(zhǎng)因子-1未見(jiàn)異常；左手X線正位片示：過(guò)度生長(zhǎng)，骨齡發(fā)育正常(圖1)。睡眠腦電圖、頭顱MRI、聽力檢查、腹部和心臟超聲未見(jiàn)異常。FMR1基因CGG重復(fù)次數(shù)正常；染色體基因芯片分析(CytoScan HD芯片，Affymetrix，USA)未發(fā)現(xiàn)致病性拷貝數(shù)變異(CNVs)。

圖1 患兒左手X線正位片

Fig 1 Anterioposterior radiograph of the patient′s left hand

Notes Overgrowth but normal bone age

孤獨(dú)癥診斷會(huì)談問(wèn)卷(ADI-R)：社會(huì)互動(dòng)異常15分(截止分10分)，溝通異常17分(截止分8分)，局限、重復(fù)、刻板的行為模式3分(截止分3分)，36月齡前發(fā)展異常的跡象3分(截止分1分)。孤獨(dú)癥診斷觀察量表(ADOS)：溝通7分[孤獨(dú)癥截止分4分，孤獨(dú)癥譜系障礙(ASD)截止分2分]，相互性社會(huì)互動(dòng)7分(孤獨(dú)癥截止分7分，ASD截止分4分)，游戲1分，刻板行為和局限興趣2分。Gesell發(fā)育評(píng)估：粗動(dòng)作發(fā)育齡30月齡，發(fā)育商76；細(xì)動(dòng)作發(fā)育齡22.5月齡，發(fā)育商57；應(yīng)物能發(fā)育齡25月齡，發(fā)育商64；言語(yǔ)能23月齡，發(fā)育商59；應(yīng)人能30月齡，發(fā)育商76。診斷為ASD。

經(jīng)患兒家長(zhǎng)同意和我院倫理委員會(huì)審核，采用EDTA抗凝管抽取患兒及其父母靜脈血各2 mL，并抽提基因組DNA(QIAamp DNA提取試劑盒，QIAGEN公司)。提取的DNA用DNA酶片段化后用磁珠法純化，隨后進(jìn)行PCR擴(kuò)增并連接上接頭序列，經(jīng)TruSight One Sequencing Panel(llumina Inc，美國(guó))2次捕獲及純化，再經(jīng)PCR擴(kuò)增和純化，獲得的最終文庫(kù)在MiSeq測(cè)序儀(illumina Inc，USA)上進(jìn)行測(cè)序。其中TruSight One Sequencing Panel根據(jù)人類基因突變數(shù)據(jù)庫(kù)(HGMD Professional)、在線人類孟德爾遺傳數(shù)據(jù)庫(kù)(OMIM)、GeneTests網(wǎng)站 (www.genetests.org)、Illumina等公司的其他商業(yè)化試劑盒的信息，納入了4 813個(gè)與已知臨床表型相關(guān)的基因，其中包含102個(gè)ASD相關(guān)基因(http://www.illumina.com/products/trusight-one-sequencing-panel.ilmn)。

實(shí)驗(yàn)產(chǎn)生1.0 Gb數(shù)據(jù)，平均測(cè)序深度為114.8×，10×以上的捕獲區(qū)覆蓋率達(dá)98.10%，平均插入片段大小237 bp，共檢測(cè)到變異8 982個(gè)。經(jīng)篩選流程篩選(圖2)，并結(jié)合患兒臨床資料和生物信息學(xué)軟件預(yù)測(cè)結(jié)果，對(duì)各個(gè)基因的功能、變異情況以及遺傳模式進(jìn)行分析發(fā)現(xiàn)，ASD相關(guān)的EHMT1、PCDH9、NLGN4X的錯(cuò)義突變、CHD8的無(wú)義突變(c.307C>T, p.Gln103*)有致病可能(表1)。圖3為二代高通量測(cè)序發(fā)現(xiàn)患兒CHD8基因存在突變。

圖2 高通量數(shù)據(jù)篩選流程圖

Fig 2 The filtering process of high-throughput sequencing data in this study

Notes Internal database of KingMed Center for Clinical Laboratory Co., Ltd.

表1 本文病例ASD相關(guān)基因候選變異總結(jié)Tab 1 Summary of candidate causative variants in ASD related genes in this patient

Notes 1)1000 Genome Project

圖3 本文病例二代高通量測(cè)序發(fā)現(xiàn)CHD8基因存在突變

Fig 3CHD8 mutation was identified via next-generation sequencing

Sanger測(cè)序的PCR引物序列及退火溫度如表2所示。該家系Sanger測(cè)序結(jié)果(圖4)顯示，患兒EHMT1、PCDH9的突變來(lái)源于父親，NLGN4X的突變來(lái)源于母親。雙親未發(fā)現(xiàn)CHD8的突變，患兒攜帶的可能為新發(fā)突變，不能除外其父母為性腺嵌合體的可能。

表2 本文病例可疑致病基因外顯子編碼區(qū)引物序列及退火溫度Tab 2 PCR primers and Tm used in this study

2 文獻(xiàn)復(fù)習(xí)

以“((“autistic disorder”[MeSH Terms] OR (“autistic”[All Fields] AND “disorder”[All Fields]) OR “autistic disorder”[All Fields] OR “autism”[All Fields]) OR (“autistic disorder”[MeSH Terms] OR (“autistic”[All Fields] AND “disorder”[All Fields]) OR “autistic disorder”[All Fields] OR “autistic”[All Fields])) AND CHD8[All

圖4 本文病例及其父母可疑致病基因突變位點(diǎn)Sanger測(cè)序圖

Fig 4 Sanger sequencing validation of candidate causative variants in this family

Notes A:CHD8，c.307C>T, p.Gln103* was found in proband; B:EHM1, c.2970G>7,9.Gln990His were found in proband and his father; C:PCDH9, c.2886C>G,p.His962Gln were found in proband and his father; D:NLGN4X, c.1486G>A, p.Val496Ile were found in proband and his mother

Fields]”為檢索式檢索PubMed數(shù)據(jù)庫(kù)，以檢索詞“CHD8”與“孤獨(dú)癥”檢索萬(wàn)方數(shù)據(jù)庫(kù)和中國(guó)知網(wǎng)，檢索時(shí)間均為建庫(kù)至2015年1月31日，共檢索到英文文獻(xiàn)13篇、中文文獻(xiàn)9篇；排除其中不相關(guān)文獻(xiàn)10篇，12篇文獻(xiàn)中關(guān)于“CHD8基因突變與ASD”的臨床相關(guān)研究3篇。

Bernier等[1]通過(guò)對(duì)3 730例臨床診斷為發(fā)育遲緩(DD)或ASD的患者進(jìn)行CHD8基因測(cè)序，并結(jié)合O′Roak的研究小組前期外顯子測(cè)序(209例ASD患者)[2]和靶基因檢測(cè)(2 446例ASD患者)[3]的結(jié)果，匯總了CHD8已知的與神經(jīng)發(fā)育障礙有關(guān)的15個(gè)重要突變(表3)，而在累計(jì)近9 000名正常對(duì)照中未發(fā)現(xiàn)CHD8基因的截?cái)嗤蛔?包括無(wú)義突變、錯(cuò)義突變、移碼突變、剪接突變、整碼突變等)。本文患兒的CHD8基因無(wú)義突變c.307C>T, p.Gln103*是新發(fā)現(xiàn)的突變，為首次報(bào)道。對(duì)既往Bernier等[1]和O′Roak等[2,3]已報(bào)道的15例CHD8基因突變患者的臨床表型進(jìn)行總結(jié)，結(jié)果如表4所示。本文患兒大頭畸形、前額突出，生長(zhǎng)較同齡兒童快，有典型的ASD表現(xiàn)，語(yǔ)言、運(yùn)動(dòng)和智力落后于同齡兒童，且有慢性便秘病史，其臨床表型與表4描述的臨床表型較為一致。

表3 已知與神經(jīng)發(fā)育障礙有關(guān)的CHD8基因重要突變

Tab 3Summary of CHD8 mutations associated with neurodevelopmental disorders (from 5′ to 3′)

ProbandSexMUT3)HGVS4)Inheritance12714.p11)MNsp.Ser62Xdenovo13986.pl1)MFsp.Tyr747XdenovoNij0234862)FFsp.Val984XInheritedmaternalAPP_109580-1002)MNsp.Glu1114Xdenovo11654.p11)FSpc.3519-2A>Gdenovo13844.p11)MNsp.Gln1238Xdenovo14016.p11)MNsp.Arg1337XdenovoNij07-066462)MMnsp.Arg1797GlnInheritedpaternalTroina26592)MFsp.Glu1932SerfsX3denovo12991.p11)MFsp.Glu2103ArgfsX3denovo12752.p11)FFsp.Leu2120ProfsX6denovoTroina20372)FFsp.Glu2136argfsX6denovoNij0108782)MAap.Lys2287delUnknown14233.p11)MFsp.Asn2371LysfsX2denovo14406.p11)MAap.His2498deldenovoOurcaseMNsp.Gln307Xdenovo

Notes MUT:mutation. 1) Patient′s mutation was previously reported by O′Roak et al[2,3];2) Patient′s mutation was previously reported by Bernier et al[1];3) Ns: nonsense; Fs: Frameshift; Sp: splice; Mns: missense-near-splice site; Aa: single amino-acid deletion;4) HGVS: human genome variant sequence

3 討論

ASD的臨床表型和基因型十分復(fù)雜，迄今確切發(fā)病機(jī)制尚不清楚，但已明確遺傳因素在其中發(fā)揮重要的作用。

表4 CHD8突變患者的臨床表型[%(n/N)]

Tab 4 Brief description of phenotypic features of patients with CHD8 mutation[%(n/N)]

PhenotypicfeaturesPositiveOurcaseNeurodevelopmentalabnormalityASD87(13/15)+Intellectualdisability60(9/15)+Delayedspeech60(9/15)+Regression47(7/15)-OvergrowthMacrocephaly80(12/15)+Tall86(12/14)+Overweight21(3/14)+DistinctfacesProminentsupraorbitalridge67(6/9)-Broadforehead44(4/9)+Hypertelorism67(6/9)-Down-slantedpalpebralfissures67(6/9)-Pointedchin44(4/9)-Largeears,fleshyearlobes44(4/9)-GastrointestinalproblemsChronicconstipation,intermittentdiar-rhea80(12/15)+SleepproblemsDifficultiesinfallingasleep,frequentnight-timeawakenings67(10/15)-BehavioralabnormalityAttentionalproblems,anxietyproblems73(11/15)+MotordelayGrossmotordelay,impairedfinemotorcoordinationskills67(10/15)+OthersCesareansection33(5/15)+Birthinduction/augmentation40(6/15)-

Notes +: present; -: absent

將異質(zhì)性明顯的疾病根據(jù)某些特征分為不同的亞型，可提高研究對(duì)象的同質(zhì)性，有利于病因的發(fā)現(xiàn)和致病機(jī)制的闡述。既往研究發(fā)現(xiàn)，ASD群體中大頭圍(>同年齡同性別P97)的比例為11%～30%[4～6]，高于正常人群，且頭圍與ASD的某些臨床表型如社交障礙的嚴(yán)重程度、智力低下等似乎存在聯(lián)系[7, 8]，提示可將大頭圍作為ASD的一種分型標(biāo)準(zhǔn)。

近年來(lái)從亞型的角度分析，已發(fā)現(xiàn)某些基因(如PTEN[9, 10]、CHD8[1]、RAB39B[11]等)在伴大頭畸形的ASD患者發(fā)病中起重要作用。本文患兒大頭畸形、生長(zhǎng)過(guò)快，有明顯的ASD特征，伴隨運(yùn)動(dòng)發(fā)育落后和慢性便秘，F(xiàn)MR1基因CGG重復(fù)次數(shù)正常，初步排除了脆性X染色體綜合征，染色體基因芯片分析未發(fā)現(xiàn)致病性CNVs的存在。為明確診斷，運(yùn)用二代高通量測(cè)序方法對(duì)4 813個(gè)臨床相關(guān)基因的外顯子區(qū)進(jìn)行測(cè)序，重點(diǎn)關(guān)注與ASD有關(guān)的基因，發(fā)現(xiàn)患兒EHMT1、PCDH9、NLGN4X、CHD8基因存在突變，其中EHMT1、PCDH9來(lái)源于未患病父親，且dbSNP及千人基因組數(shù)據(jù)庫(kù)有收錄(EHMT1：rs143669310，0.0014；PCDH9：rs192811737，0.0018)，不考慮為患兒的致病原因。NLGN4X的突變來(lái)源于母親，該基因突變引起的ASD可認(rèn)為是X連鎖隱性遺傳，但未見(jiàn)報(bào)道過(guò)度生長(zhǎng)、大頭畸形等表型。結(jié)合本文患兒的臨床表型認(rèn)為NLGN4X的變異可能并不是其致病原因，而根據(jù)既往關(guān)于CHD8基因的文獻(xiàn)報(bào)道[1～3]，CHD8雜合無(wú)義突變c.307C>T,p.Gln103*(NM_020920.3)使編碼產(chǎn)物變短而破壞蛋白質(zhì)原有功能，更有可能是患兒ASD相關(guān)的重要遺傳學(xué)改變。

CHD8基因位于14q11.2區(qū)，總長(zhǎng)約46 kb，由37個(gè)外顯子組成；其編碼產(chǎn)物通過(guò)結(jié)合β-catenin募集組蛋白H1，改變?nèi)旧|(zhì)空間結(jié)構(gòu)而發(fā)揮轉(zhuǎn)錄抑制作用，是Wnt/β-catenin信號(hào)通路重要的調(diào)節(jié)物[12, 13]，而該通路對(duì)神經(jīng)元增殖和分化、突起生長(zhǎng)和突觸形成作用較大[14～18]，與ASD發(fā)病顯著相關(guān)[19～21]。O′Roak等于2012年首次對(duì)209例ASD患者及其未患病同胞進(jìn)行外顯子組測(cè)序，發(fā)現(xiàn)2例患者存在CHD8基因突變[2]；隨后相關(guān)研究已證實(shí)，CHD8新發(fā)的截?cái)嗤蛔兪茿SD發(fā)病的重要危險(xiǎn)因素[1, 3, 22]。此外，Talkowski等[23]在攜帶染色體平衡易位的ASD患者中發(fā)現(xiàn)了CHD8的破壞，也提示了該基因?qū)τ贏SD發(fā)病的重要作用。

Sugathan等[24]在神經(jīng)前體細(xì)胞模擬CHD8基因功能性的雜合缺失發(fā)現(xiàn)，CHD8基因調(diào)節(jié)許多功能各異但與ASD相關(guān)的基因(SCN2A、DLG2、SHANK3、MBD3等)，以及與突觸形成、神經(jīng)元分化、細(xì)胞黏附等有關(guān)的基因(LAMA4、NCAM1、MEGF10等)。突變的CHD8可通過(guò)影響上述基因而影響神經(jīng)系統(tǒng)的正常發(fā)育。chd8基因敲除的斑馬魚模型再現(xiàn)了大頭畸形[1, 24]和胃腸道動(dòng)力不足[1]的臨床表型。通過(guò)神經(jīng)元特殊染色發(fā)現(xiàn)，前腦和中腦神經(jīng)元前體細(xì)胞的過(guò)度增生導(dǎo)致這2個(gè)部位體積增大，造成頭部過(guò)度生長(zhǎng)；而發(fā)育過(guò)程中腸道神經(jīng)叢增殖或是遷移障礙導(dǎo)致腸道神經(jīng)元減少，影響正常的胃腸道功能。細(xì)胞和動(dòng)物研究結(jié)果均說(shuō)明CHD8破壞會(huì)導(dǎo)致神經(jīng)元發(fā)育的缺陷。

針對(duì)CHD8基因異常的ASD患者，除通過(guò)特殊教育和訓(xùn)練提高認(rèn)知、社交和適應(yīng)能力外，還應(yīng)關(guān)注患者的胃腸道問(wèn)題和睡眠狀況。此外，既往研究表明，CHD8與胃腸道、皮膚等部位多種腫瘤密切相關(guān)[25～28]；Bernier的研究中1例41歲的CHD8突變患者合并腫瘤，另1例將突變傳給患兒的父親罹患直腸癌[1]，這提示注意腫瘤的早期征象可能是CHD8突變患者日后定期隨訪的又一重點(diǎn)。

總之，CHD8基因是ASD重要的易感基因；ASD患兒合并大頭畸形、生長(zhǎng)過(guò)快，伴有慢性便秘或間歇性腹瀉時(shí)應(yīng)考慮存在CHD8基因突變可能，可行基因檢測(cè)協(xié)助診斷。

[1]Bernier R, Golzio C, Xiong B, et al. Disruptive CHD8 mutations define a subtype of autism early in development. Cell, 2014, 158(2): 263-276

[2]O′Roak BJ, Vives L, Girirajan S, et al. Sporadic autism exomes reveal a highly interconnected protein network of de novo mutations. Nature, 2012, 485(7397): 246-250

[3]O′Roak BJ, Vives L, Fu W, et al. Multiplex targeted sequencing identifies recurrently mutated genes in autism spectrum disorders. Science, 2012, 338(6114): 1619-1622

[4]Woodhouse W, Bailey A, Rutter M, et al. Head circumference in autism and other pervasive developmental disorders. J Child Psychol Psychiatry, 1996, 37(6): 665-671

[5]van Daalen E, Swinkels SH, Dietz C, et al. Body length and head growth in the first year of life in autism. Pediatr Neurol,2007,37(5):324-330

[6]Lainhart JE, Bigler ED, Bocian M, et al. Head circumference and height in autism: a study by the Collaborative Program of Excellence in Autism. Am J Med Genet A, 2006, 140(21): 2257-2274

[7]Davis JM, Keeney JG, Sikela JM, et al. Mode of genetic inheritance modifies the association of head circumference and autism-related symptoms: a cross-sectional study. PLoS One, 2013, 8(9): e74940

[8]Sacco R, Militerni R, Frolli A, et al. Clinical, morphological, and biochemical correlates of head circumference in autism. Biol Psychiatry, 2007, 62(9): 1038-1047

[9]Marchese M, Conti V, Valvo G, et al. Autism-epilepsy phenotype with macrocephaly suggests PTEN, but not GLIALCAM, genetic screening. BMC Med Genet,2014,15:26

[10]Klein S, Sharifi-Hannauer P, Martinez-Agosto JA. Macrocephaly as a clinical indicator of genetic subtypes in autism. Autism Res, 2013, 6(1): 51-56

[11]Giannandrea M, Bianchi V, Mignogna ML, et al. Mutations in the small GTPase gene RAB39B are responsible for X-linked mental retardation associated with autism, epilepsy, and macrocephaly. Am J Hum Genet, 2010, 86(2): 185-195

[12]Thompson BA, Tremblay V, Lin G, et al. CHD8 is an ATP-dependent chromatin remodeling factor that regulates beta-catenin target genes. Mol Cell Biol, 2008, 28(12): 3894-3904

[13]Nishiyama M, Skoultchi AI, Nakayama KI. Histone H1 recruitment by CHD8 is essential for suppression of the Wnt-beta-catenin signaling pathway. Mol Cell Biol, 2012, 32(2): 501-512

[14]Machon O, Backman M, Machonova O, et al. A dynamic gradient of Wnt signaling controls initiation of neurogenesis in the mammalian cortex and cellular specification in the hippocampus. Dev Biol, 2007, 311(1): 223-237

[15]Zhou CJ, Borello U, Rubenstein JL, et al. Neuronal production and precursor proliferation defects in the neocortex of mice with loss of function in the canonical Wnt signaling pathway. Neuroscience, 2006, 142(4): 1119-1131

[16]Salinas PC. Wnt signaling in the vertebrate central nervous system: from axon guidance to synaptic function. Cold Spring Harb Perspect Biol, 2012, 4(2)[17]Farias GG, Godoy JA, Cerpa W, et al. Wnt signaling modulates pre- and postsynaptic maturation: therapeutic considerations. Dev Dyn, 2010, 239(1): 94-101

[18]Speese SD, Budnik V. Wnts: up-and-coming at the synapse. Trends Neurosci, 2007, 30(6): 268-275

[19]Krumm N, O′Roak BJ, Shendure J, et al. A de novo convergence of autism genetics and molecular neuroscience. Trends Neurosci, 2014, 37(2): 95-105

[20]Okerlund ND, Cheyette BN. Synaptic Wnt signaling-a contributor to major psychiatric disorders?. J Neurodev Disord, 2011, 3(2): 162-174

[21]Hormozdiari F, Penn O, Borenstein E, et al. The discovery of integrated gene networks for autism and related disorders. Genome Res, 2015, 25(1): 142-154

[22]Neale BM, Kou Y, Liu L, et al. Patterns and rates of exonic de novo mutations in autism spectrum disorders. Nature, 2012, 485(7397): 242-245

[23]Talkowski ME, Rosenfeld JA, Blumenthal I, et al. Sequencing chromosomal abnormalities reveals neurodevelopmental loci that confer risk across diagnostic boundaries. Cell, 2012, 149(3): 525-537

[24]Sugathan A, Biagioli M, Golzio C, et al. CHD8 regulates neurodevelopmental pathways associated with autism spectrum disorder in neural progenitors. Proc Natl Acad Sci U S A, 2014, 111(42): E4468-E4477

[25]Sawada G, Ueo H, Matsumura T, et al. CHD8 is an independent prognostic indicator that regulates Wnt/beta-catenin signaling and the cell cycle in gastric cancer. Oncol Rep, 2013, 30(3): 1137-1142

[26]Tahara T, Yamamoto E, Madireddi P, et al. Colorectal carcinomas with CpG island methylator phenotype 1 frequently contain mutations in chromatin regulators. Gastroenterology, 2014, 146(2): 530-538

[27]Lawrence MS, Stojanov P, Mermel CH, et al. Discovery and saturation analysis of cancer genes across 21 tumour types. Nature, 2014, 505(7484): 495-501

[28]Kim MS, Chung NG, Kang MR, et al. Genetic and expressional alterations of CHD genes in gastric and colorectal cancers. Histopathology, 2011, 58(5): 660-668

(本文編輯：丁俊杰)

《中國(guó)循證兒科雜志》主要欄目及其特點(diǎn)

述評(píng)：在高度精練又不失全面地評(píng)價(jià)某一臨床或基礎(chǔ)研究熱點(diǎn)問(wèn)題的基礎(chǔ)上，前瞻性地指出研究可能的切入點(diǎn)和方向。

專家對(duì)談錄：是國(guó)內(nèi)外或境內(nèi)外專家共同參與的談話式欄目。選題為最新的基礎(chǔ)或臨床研究問(wèn)題。

論著：本刊論著文稿的內(nèi)容應(yīng)遵循以下類型醫(yī)學(xué)研究報(bào)告規(guī)范撰寫。

隊(duì)列研究、病例對(duì)照研究和橫斷面研究以觀察性流行病學(xué)研究報(bào)告的質(zhì)量(Strengthening the reporting of observational studies in epidemiology，STROBE) 為標(biāo)準(zhǔn)，STROBE原文見(jiàn)www.strobe-statement.org。STROBE中文解讀見(jiàn)《中國(guó)循證兒科雜志》，2010，5(3)：223-227

觀察性研究的Meta 分析(Meta-analysis of observational studies in epidemiology，MOOSE)報(bào)告規(guī)范原文見(jiàn)JAMA,2000, 283(15): 2008-2012。隨機(jī)對(duì)照試驗(yàn)Meta分析報(bào)告質(zhì)量(the quality of reporting of Meta-analysis of randomized controlled trial, QUORPM)原文見(jiàn)Lancet,1999,354(9193):1896-1900。MOOSE和QUOROM中文解讀見(jiàn)《中國(guó)循證兒科雜志》，2010，5(1)：60-63

非劣效性和等效性隨機(jī)對(duì)照試驗(yàn)的報(bào)告規(guī)范(Consolidated standards of reporting trials,CONSORT)原文見(jiàn)http://www.consort-statement.org。CONSORT中文解讀見(jiàn)《中華流行病學(xué)雜志》，2006，12(27)：1086-1088

非隨機(jī)對(duì)照設(shè)計(jì)報(bào)告規(guī)范(Transparent reporting of evaluations with nonrandomized designs，TREND)原文見(jiàn)Am J Public Health,2004,94(3):361-366。TREND中文解讀見(jiàn)《中華流行病學(xué)雜志》，2006，4(27)：408-410

診斷試驗(yàn)準(zhǔn)確性研究的報(bào)告規(guī)范(Standards for reporting of diagnostic accuracy,STARD)原文見(jiàn)www.stard-statement.org。STARD中文解讀見(jiàn)《中華流行病學(xué)雜志》，2006，10(27)：910-912

隨機(jī)對(duì)照試驗(yàn)報(bào)告規(guī)范(CONSORT)原文見(jiàn)http://www.consort-statement.org。CONSORT中文解讀見(jiàn)《中國(guó)循證兒科雜志》，2010，5(2)：146-150

遺傳關(guān)聯(lián)性研究報(bào)告規(guī)范(STREGA)原文見(jiàn)http://www.strega-statement.org。STREGA中文解讀見(jiàn)《中國(guó)循證兒科雜志》，2010，5(4)：304-307

病例報(bào)告：應(yīng)遵循病例報(bào)告的報(bào)告規(guī)范(CARE)撰寫，原文見(jiàn)http://www.care-statement.org/index.html。CARE中文解讀見(jiàn)《中國(guó)循證兒科雜志》，2014，9(3)：216-219

A novel CHD8 gene mutation in a patient with autism spectrum disorder and literature review

ZHENGQing-wen1,ZHANGuo-dong1,TANGChun1,BIXin2,ZHAOWei-wei2,ZOUXiao-bing1

(1TheThirdAffiliatedHospitalofSunYat-senUniversity,Guangzhou510630; 2KingMedCenterforClinicalLaboratoryCo.,Ltd.,Guangzhou510330,China)

ZOU Xiao-bing，E-mail:zouxb@vip.tom.com; ZHAO Wei-wei，E-mail:lab-zhaoweiwei@kingmed.com.cn

ObjectiveTo draw attention to the phenotype and genotype of patients with autism spectrum disorders (ASD) carryingCHD8 mutation.MethodsThe clinical data of one patient with ASD carryingCHD8 mutation, including clinical manifestations, laboratory findings, genetic testing results and family investigation were summarized and analyzed, and relevant literatures aboutCHD8 mutation were reviewed in this article.Results①The 3-year-and-3-month boy was presented with macrocephaly (head circumference was 55.0 cm,Z>4s), overgrowth (height was 107.0 cm,Z>2s; weight was 23.5 kg,Z>2s) and typical autistic behavior as well as significant motor delay and gastrointestinal(GI) problems (chronic constipation). No additional patient was found in his family. ②Targeting 4 813 genes associated with known clinical phenotypes, a total of 8 982 variances were found in the patient via next-generation high-throughput DNA sequencing. Missense mutations of autism-related genesEHMT1,PCDH9,NLGN4Xand nonsense mutation ofCHD8 [c.307C>T, p.Gln103*(NM_020920.3)] were probably pathogenic after filtering unrelated variances.EHMT1,PCDH9 mutations were identified in his father andNLGN4Xmutation in his mother, butCHD8 mutation was not identified in his parents via Sanger sequencing. ③To date a total of 15 independentCHD8 mutations associated with neurodevelopmental disorders has been reported worldwide. Common phenotypic features of patients withCHD8 disruptions include autism(87%), overgrowth(86%), macrocephaly(80%), GI complaints(80%), motor delay(67%, especially fine motor coordination) and sleep problems(67%).ConclusionThe mutation ofCHD8 is an important risk factor of ASD. ASD patients with macrocephaly, overgrowth and GI problems should be considered as carryingCHD8 mutation and genetic testing is recommended to confirm the diagnosis.

Autism spectrum disorder ;CHD8; Macrocephaly; Overgrowth; Next-generation sequencing

國(guó)家重點(diǎn)基礎(chǔ)研究發(fā)展計(jì)劃(973計(jì)劃)：2012CB517901

1 中山大學(xué)附屬第三醫(yī)院 廣州，510630; 2 廣州金域醫(yī)學(xué)檢驗(yàn)中心 廣州,510330

鄒小兵，E-mail:zouxb@vip.tom.com；趙薇薇，E-mail:lab-zhaoweiwei@kingmed.com.cn

10.3969/j.issn.1673-5501.2015.03.012

2015-03-03

2015-05-27)