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北大学者发明新技术助力胚胎植入前无创检测

时间:2019-07-04 17:00来源:未知 作者:admin 点击:
由于生育年龄推迟、社会压力增大、不良生活方式等因素,一部分人遭遇了难以自然生育的困境,继而发展了人类辅助生殖技术(Assisted Reproductive Technology,ART),其代表的技术,如试

  由于生育年龄推迟、社会压力增大、不良生活方式等因素,一部分人遭遇了难以自然生育的困境,继而发展了人类辅助生殖技术(Assisted Reproductive Technology,ART),其代表的技术,如试管婴儿技术被誉为20世纪医学界的一大奇迹,给许多无法自然受孕的夫妇带去了希望。

  然而要想成功妊娠,孕育出健康的胎儿,胚胎移植前基因检测是不可或缺的。其中,胚胎植入前染色体非整倍性检测(PGT-A)是指在胚胎植入之前利用多种技术检查早期胚胎的染色体数目和结构是否正常。

  目前,临床上最常用的染色体非整倍性检测方法是取用胚胎滋养外胚层的细胞进行组织活检。尽管利用该方法筛选出整倍体胚胎后能够提高胚胎移植的成功率,但关于这一方法的准确性、安全性以及适宜的人群,始终存在争议。

  6月24日,北京大学生物医学前沿创新中心谢晓亮教授及其合作者在美国《国家科学院院刊》(PNAS)上发文称,基于严格的方法,比较植入前无创基因检测和滋养外胚层活检两种方法对染色体非整倍性进行检测,最终证明利用胚胎培养液游离DNA的无创基因检测(niPGT-A)比滋养外胚层活检(TE)具有更高的准确性和可靠性。

  中国人民解放军总医院(301医院)生殖中心姚元庆主任表示,“谢晓亮教授和Racowsky教授的工作为植入前胚胎无创检测的临床转化奠定了坚实的理论基础,在这些工作的基础上,开展基于二代测序的胚胎培养液游离DNA非整倍体检测的多中心、大样本、随机对照的临床试验将为无创植入前胚胎检测的临床应用提供循证医学证据,最终将这一创新技术应用于临床实践成为可能。”

  整倍体/非整倍体嵌合体可能导致滋养外胚层活检结果不准确,图片来自PNAS

  胚胎滋养外胚层活检的准确性与胚胎嵌合体相关。嵌合体指的是在同一个胚胎中存在两种及以上的不同基因型的细胞。在胚胎中,多达30-40%的人类胚泡是嵌合体,其中整倍体/非整倍体嵌合体的发生率在2.0-2.9%和14.0%-17.3%之间,这将不可避免地导致滋养外胚层活检的结果不能反映内细胞团以及其最终形成的胎儿的染色体整倍性。

  除了对准确性的担忧外,滋养外胚层活检的安全性也值得关注。虽然目前尚未有证据明确指出滋养外胚层活检对胚胎有害,但其安全性仍然是未知的。因为取出滋养外胚层细胞可能会对胚胎带来潜在的伤害,甚至影响胚胎的着床能力。

  由于滋养外胚层活检存在上述局限性,2016年谢晓亮及其合作者首次开发出了一种无创的植入前非整倍体基因检测方法(niPGT-A),利用胚胎培养液中游离的DNA进行基因检测。但由于结果一致性较差,这种方法当时未被看好,并且被认为不如传统的滋养外胚层活检技术。

  在这项研究中,研究人员首先收集了52个冷冻捐赠的人类囊胚,这些胚囊之前曾经过外部实验室的滋养外胚层活检测试。随后研究人员将这些囊胚解冻后,在微滴培养基中单独培养24小时,收集培养基和胚胎,扩增并测序。

  之后,研究者对比了利用培养液游离 DNA的无创基因检测和滋养外胚层活检得到的染色体倍性结果,以及对整个胚胎测序得到的真实结果。通过使用严格的程序来设置嵌合体阈值,研究者优化了无创基因检测结果的假阴性率和假阳性率,其阳性预测值和特异性均远高于滋养外胚层活检。此外,无创基因检测的胚胎倍性和染色体拷贝数的一致率均高于滋养外胚层活检。

  这些结果表明,无创基因检测不易发生与胚胎镶嵌相关的错误,并且比滋养外胚层活检更可靠。

  接下来,研究者将继续改进该方法,提升准确性。未来该方法将有望取代滋养外胚层活检技术,为使用试管婴儿技术的夫妇带来更为安全可靠的胚胎植入前检测。

  2019年6月24日,《美国科学院院报》在线发表了谢晓亮教授和哈佛大学布列根和妇女医院(Brigham and Women’s Hospital)Catherine Racowsky教授团队完成的无创植入前胚胎检测的最新研究成果,采用二代测序技术,该研究检测了42个捐赠的整个胚胎的非整倍性,并分别与同一胚胎的胚胎滋养层活检的二代测序结果和胚胎培养液游离DNA的二代测序结果进行了比较。

  结果显示,胚胎培养液游离DNA检测比胚胎活检更准确地代表胚胎的非整倍性。这一研究结果的发表对于无创植入前胚胎检测技术的临床应用具有重要意义。

  多年来,通过胚胎的非整倍性检测,选择具有发育和种植潜能的胚胎进行移植,提高体外受精和胚胎移植的成功率,降低多胎率的发生是辅助生殖技术的热点问题,也存在一定的争议,主要是胚胎活检造成损伤的安全性风险问题以及胚胎嵌合造成的几个活检胚胎细胞的检测结果不能反映整个胚胎整倍性的问题。

  2016年,谢晓亮教授的团队在《美国科学院院报》上报告了首例基于胚胎培养液游离DNA二代测序的植入前胚胎检测的“试管婴儿”出生。在无创植入前胚胎检测的探索上走出了第一步,在避免胚胎活检所造成的可能的安全性问题上提供了解决方案,但源自卵丘细胞或是颗粒细胞母源DNA和胚胎嵌合对检测结果的干扰,仍然是无创植入前胚胎检测的瓶颈问题。

  在谢晓亮教授和Racowsky教授的最新研究中,通过改进单精子卵胞浆显微注射前的卵母细胞脱颗粒细胞技术和胚胎培养液收集时间的调整,基本解决了母源DNA对检测结果的干扰,并通过以整个胚胎检测结果作为标准的方法,发现胚胎培养液游离DNA检测与整个胚胎检测结果的符合程度显著高于胚胎活检的结果,从而为回答胚胎培养液游离DNA检测的准确性问题,提供了有力证据。

  我们认为,无创植入前胚胎的发育和着床潜能检测将全面改变辅助生殖技术的临床策略。

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n={captchaFlag:!0,captchaApp:e.getConfig(captchaAppId,i360),smDeviceId:i()};t=a.extend(n,t),t.password=r(t.password),t.passwordAgain=r(t.passwordAgain)t.password,o(t,{emailActiveFlag:loginEmailActiveFlag,passwordAgain:rePassword,smsToken:smscode,nickname:nickName,username:userName,agreeLicence:is_agree});var s=new c(t,{},!0);return s.post(s.I360+/reg/doregAccount).done(function(){u()})},signIn:function(t){var n={o:sso,m:login,lm:mobile==t.type?1:0,captFlag:1,rtype:data,validatelm:e.getConfig(signIn.mobile.isMustUseMobileSignIn,!1)?1:0,isKeepAlive:!1,captchaApp:e.getConfig(captchaAppId,i360),userName:t.account,smDeviceId:i()};returnmobile==t.type?t.acctype=2:t.password=r(t.password),a.when().then(function(){return t.tokene.sync.getToken(t.account).done(function(e){t.token=e.token})}).then(function(){return(new c(a.extend(n,t),{},!0)).post().done(function(){u()})}).then(function(t){return e.sync.setCookie(t.s)}).then(function(){return e.getUserInfo(!1)})},signOut:function(t){var n=e.getConfig(supportHttps,l),r=https==e.getConfig(protocol,null).toLowerCase();void 0===t!0===t?t=e.getConfig(domainList,[]):a.isArray(t)(t=[t]);var i,s=[];return a.each(t,function(e,t){a.inArray(t,n)>

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