Understanding the impact of ovarian stimulation on oocyte and embryo quality by tandem RNA and protein expression analysis of oocytes and preimplantation embryonic stages

通过卵母细胞和植入前胚胎阶段的串联 RNA 和蛋白质表达分析了解卵巢刺激对卵母细胞和胚胎质量的影响

• 批准号: 279728933
• 负责人: Privatdozent Dr. Michele Boiani
• 金额: --
• 依托单位: Institut für Biostatistik und Informatik in Medizin und Alternsforschung
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/279728933?language=en
• 关键词: Understanding; impact; ovarian; stimulation; oocyte

Ovarian stimulation with gonadotropins is a key component of assisted reproductive technologies (ARTs), aimed at increasing the number of oocytes and embryos, and, thereby, the chance of establishing a pregnancy. However, independent studies suggest that ovarian stimulation has detrimental effects on oocyte and embryo quality. Indeed, ovarian stimulation has been shown to lead to a significantly lower proportion of normal embryos, as well as decreased implantation and post-implantation development rates. Here, we aim to combine high-throughput transcriptomics and proteomics with statistical and computational modeling in a holistic approach, to analyze preimplantation development and understand the effects of ovarian stimulation on oocyte and embryo quality in the mouse model of human reproduction. The mouse has contributed more than any other model to the advancement of ARTs. Experimentally, we will quantitatively measure RNA, mRNA poly(A) tail lengths, and protein using RNA-seq, PAL-seq, and SILAC LC-MS/MS, respectively, across the mouse oocyte and derived embryos at seven different developmental stages (1-cell, 2-cell (early and late), 4-cell, 8-cell, morula and blastocyst), with and without a background of ovarian stimulation. Computationally, we will develop and apply novel statistical methods and network-based approaches to analyze these comprehensive data and describe the gene expression cascade in early embryonic development, with and without a background of ovarian stimulation. In particular, we will propose several regression methods to (1) describe protein expression levels as a function of concomitant and precedent mRNA levels, as well as other sequence and functional properties of the protein/mRNA considered; (2) identify the transcription factors with crucial roles throughout embryonic development; (3) predict the targets of noncoding RNA and evaluate their contribution to the gene expression cascade; and (4) assess allele-specific expression, to get an insight into the time course of embryonic genome activation (EGA). Additionally, we will test the hypothesis that control mechanisms involving the poly(A) tail of mRNA are a main contributor to the discrepancies between transcriptomic and proteomic expression patterns during early development. Finally, we will use network biology approaches to identify gene/protein interactions that may underlie developmental progression, and exploit comparative genomics to establish the relevance of our results for human ARTs. With more and more women postponing childbearing to later stages in their lives and turning to ARTs, our study makes a fundamental contribution to understand the basic molecular and cellular aspects of early embryonic development, and to evaluate the impact of common ART treatments on oocyte and embryo quality.

促性腺激素刺激卵巢是辅助生殖技术（ARTs）的关键组成部分，旨在增加卵母细胞和胚胎的数量，从而增加怀孕的机会。然而，独立研究表明卵巢刺激对卵母细胞和胚胎质量有不利影响。事实上，卵巢刺激已被证明会导致正常胚胎比例显著降低，并降低着床和着床后的发育率。在此，我们旨在将高通量转录组学和蛋白质组学与统计和计算建模相结合，全面分析人类生殖小鼠模型的着床前发育，了解卵巢刺激对卵母细胞和胚胎质量的影响。老鼠比任何其他模型对ARTs的发展贡献都要大。在实验中，我们将分别使用RNA-seq、PAL-seq和SILAC LC-MS/MS定量测量小鼠卵母细胞和衍生胚胎在7个不同发育阶段（1细胞、2细胞（早期和晚期）、4细胞、8细胞、桑葚胚和囊胚）的RNA、mRNA poly(A)尾巴长度和蛋白质，以及有无卵巢刺激背景。在计算方面，我们将开发和应用新颖的统计方法和基于网络的方法来分析这些综合数据，并描述在有或没有卵巢刺激背景下早期胚胎发育中的基因表达级联。特别是，我们将提出几种回归方法来(1)将蛋白质表达水平描述为伴随和先前mRNA水平的函数，以及所考虑的蛋白质/mRNA的其他序列和功能特性；(2)确定在胚胎发育过程中起关键作用的转录因子；(3)预测非编码RNA的靶点，评估其在基因级联表达中的作用；(4)评估等位基因特异性表达，以了解胚胎基因组激活（EGA）的时间过程。此外，我们将验证涉及mRNA多(A)尾的控制机制是早期发育过程中转录组和蛋白质组表达模式差异的主要原因。最后，我们将使用网络生物学方法来识别可能导致发育进程的基因/蛋白质相互作用，并利用比较基因组学来确定我们的结果与人类art的相关性。随着越来越多的女性推迟生育到生命的后期并转向ART，我们的研究对了解早期胚胎发育的基本分子和细胞方面以及评估常见ART治疗对卵母细胞和胚胎质量的影响做出了根本性的贡献。

项目成果

An integrated genome-wide multi-omics analysis of gene expression dynamics in the preimplantation mouse embryo

• DOI: 10.1038/s41598-019-49817-3
• 发表时间: 2019-09-16
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Israel, Steffen;Ernst, Mathias;Taher, Leila
• 通讯作者: Taher, Leila

A framework for TRIM21-mediated protein depletion in early mouse embryos: recapitulation of Tead4 null phenotype over three days

• DOI: 10.1186/s12864-019-6106-2
• 发表时间: 2019-10-21
• 期刊: BMC GENOMICS
• 影响因子: 4.4
• 作者: Israel, Steffen;Casser, Ellen;Boiani, Michele
• 通讯作者: Boiani, Michele

66 Unexpected Protein Dynamics During the Oocyte-to-Embryo Transition in Mice: a Mass Spectrometry and RNA Sequencing Tandem Study

66 小鼠卵母细胞到胚胎转变过程中意外的蛋白质动态：质谱和 RNA 测序串联研究

• DOI: 10.1071/rdv30n1ab66
• 发表时间: 2017
• 期刊: Reproduction, Fertility and Development
• 作者: Israel S;Ernst M;Psathaki OE;Drexler HC;Casser E;Suzuki Y;Makalowski W;Fuellen G;Boiani M;Taher L
• 通讯作者: Taher L

O-169 Conventional ovarian stimulation with gonadotropins depletes the developmental proteome of mouse oocytes, reducing their size and compromising their fetal yield

O-169 使用促性腺激素进行常规卵巢刺激会消耗小鼠卵母细胞的发育蛋白质组，从而减小其大小并损害其胎儿产量

• DOI: 10.1093/humrep/deab127.050
• 发表时间: 2021
• 期刊: Human Reproduction
• 影响因子: 6.1
• 作者: Boiani M;Drexler HC;Fuellen G;Israel S;Makalowski W;Suzuki Y;Taher L
• 通讯作者: Taher L