Single cell expression analysis of primate-specific retroviruses-derived HPAT lincRNAs in viable human blastocysts identifies embryonic cells co-expressing genetic markers of multiple lineages.
Single cell expression analysis of primate-specific retroviruses-derived HPAT lincRNAs in viable human blastocysts identifies embryonic cells co-expressing genetic markers of multiple lineages.
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DOI：
10.1016/j.heliyon.2018.e00667
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发表时间：
2018-06
期刊：
影响因子：
4
通讯作者：
Sebastiano V
中科院分区：
文献类型：
--
作者：
Glinsky G;Durruthy-Durruthy J;Wossidlo M;Grow EJ;Weirather JL;Au KF;Wysocka J;Sebastiano V
染色体不稳定性和非整倍性在人类胚胎中非常频繁地发生，损害正常的胚胎发生，并导致细胞周期停滞，细胞活力丧失，以及50-80%的卵裂期胚胎发育失败。这种高频率的细胞灭绝事件代表了一个重大的实验障碍，挑战了从人类植入前胚胎中分离的单个细胞的分析。我们进行了单细胞表达谱的241个细胞回收32人胚胎在早期和晚期阶段的可行的人胚泡（VHB）分化。仅基于人多能性相关转录物（HPAT）的表达模式进行胚胎细胞的分类，所述人多能性相关转录物代表在人胚胎干细胞中高度表达并调节核重编程和多能性诱导的灵长类特异性转座元件衍生的lincRNA家族。然后，我们通过分析从100多个人类胚胎中回收的1，708个单个细胞和从40多个小鼠胚胎中回收的259个小鼠细胞的转录组来验证我们的发现。从VHB分化的单细胞表达分析推断的HPAT的表达引导的人胚胎发育时空重建使得能够鉴定共表达关键多能性调控基因和三种主要谱系的遗传标记的端粒酶阳性胚胎细胞。后续验证分析证实，在人类胚胎谱系分离之前，端粒酶阳性细胞共表达多个谱系的遗传标记。在这篇文章中报道的观察结果支持了一个假设，即从端粒酶阳性的前谱系细胞中产生胚胎谱系和胚外组织的发育途径表现出多谱系前体表型。
Chromosome instability and aneuploidies occur very frequently in human embryos, impairing proper embryogenesis and leading to cell cycle arrest, loss of cell viability, and developmental failures in 50–80% of cleavage-stage embryos. This high frequency of cellular extinction events represents a significant experimental obstacle challenging analyses of individual cells isolated from human preimplantation embryos. We carried out single cell expression profiling of 241 individual cells recovered from 32 human embryos during the early and late stages of viable human blastocyst (VHB) differentiation. Classification of embryonic cells was performed solely based on expression patterns of human pluripotency-associated transcripts (HPAT), which represent a family of primate-specific transposable element-derived lincRNAs highly expressed in human embryonic stem cells and regulating nuclear reprogramming and pluripotency induction. We then validated our findings by analyzing transcriptomes of 1,708 individual cells recovered from more than 100 human embryos and 259 mouse cells from more than 40 mouse embryos at different stages of preimplantation embryogenesis. HPAT's expression-guided spatiotemporal reconstruction of human embryonic development inferred from single-cell expression analysis of VHB differentiation enabled identification of telomerase-positive embryonic cells co-expressing key pluripotency regulatory genes and genetic markers of three major lineages. Follow-up validation analyses confirmed the emergence in human embryos prior to lineage segregation of telomerase-positive cells co-expressing genetic markers of multiple lineages. Observations reported in this contribution support the hypothesis of a developmental pathway of creation embryonic lineages and extraembryonic tissues from telomerase-positive pre-lineage cells manifesting multi-lineage precursor phenotype.