Noninvasive assessment of embryo implantation potential using 1H NMR metabonomics

说明(申请人提供)：多胎生育是儿童发病率和长期健康受损的一个重要原因。体外受精(IVF)通过在一个周期内移植多个胚胎，显著增加了多胎生育的数量。目前的形态标准不足以准确识别那些将成功植入的胚胎。我们基于质子核磁共振波谱的初步研究表明，对100%和0%植入病例的培养基样品记录的核磁共振波谱的多元分析表明，两组之间存在统计学上的分离(p=0.009)。此外，将核磁共振谱反卷积到存在于培养介质中的单个化合物的贡献中，表明几种化合物的浓度在100%和0%注入类之间有显著差异。基于这些全面的初步研究，我们因此假设可植入胚胎具有代谢特征，可以在条件培养液中使用1D 1H-核磁共振进行鉴定。为了验证我们的假设，我们建议建立基于核磁共振的培养基谱分析，作为一种非侵入性预测胚胎植入成功的工具。在特定的目标1，我们将开发统计模型，以预测胚胎植入成功的基础上，从一维1H核磁共振波谱记录从0%和100%植入案例。在具体目标2中，我们将通过将我们的基于核磁共振的分析方案应用于部分胚胎植入成功的病例的新介质样本来测试这些预测模型的稳健性。这将使我们能够评估模型的敏感性、特异性和预测值。这项研究的意义在于：(I)建立基于核磁共振的体外受精培养液分析，作为一种非侵入性预测胚胎植入成功的工具，以降低多次试管受精怀孕的可能性，以及(Ii)对早期胚胎发育的生物学获得新的见解。公共卫生相关性：体外受精(IVF)治疗导致多胞胎数量显著增加，严重损害了孕产妇和围产期健康，并增加了社会的医疗保健成本。必须开发非侵入性的工具，允许在每个试管受精周期中选择和移植单个胚胎。我们提出了一项初步研究，利用质子核磁共振波谱(1H-核磁共振)来表征条件培养液中的可植入胚胎的代谢特征。

DESCRIPTION (provided by applicant): Multiple births represent a significant cause of childhood morbidity and long-term compromised health. In vitro fertilization (IVF) treatment has significantly increased the number of multiple births by the transfer multiple embryos in a single cycle. Current morphological criteria are not sufficient to precisely identify those embryos that will implant successfully. Our preliminary studies based on proton nuclear magnetic resonance spectroscopy (1H-NMR) show that multivariate analysis of the NMR spectra recorded for culture media samples from 100% and 0% implantation cases revealed a statistical separation of the two groups (p=0.009). In addition, deconvolution of the NMR spectra into the contributions from individual compounds present in the culture media suggested that the concentrations of several compounds were significantly different between the 100% and 0% implantation classes. Based on these comprehensive preliminary studies, we thus hypothesize that implantable embryos have metabolic characteristics that can be identified in conditioned culture media using 1D 1H-NMR. To test our hypothesis, we propose to establish NMR-based profiling of spent culture media as a tool to non-invasively predict embryo implantation success. In Specific Aim 1 we will develop statistical models to predict embryo implantation success based on 1D 1H NMR spectra recorded for media from 0% versus 100% implantation cases. In Specific Aim 2, we will test the robustness of these predictive models by applying our protocol for NMR-based profiling to new media specimens from cases with partial embryo implantation success. This will enable us to evaluate the models' sensitivity, specificity, and predictive value. The significance of the proposed research arises from (i) establishing NMR-base profiling of IVF media as a tool to non-invasively predict embryo implantation success to reduce the likelihood of multiple IVF pregnancies, and (ii) gaining new insights into the biology of early embryo development. PUBLIC HEALTH RELEVANCE: Treatment with in vitro fertilization (IVF) has resulted in a significant increase in the number of multiple births, significantly compromising both maternal and perinatal health, and increasing health care costs to society. It is imperative to develop non-invasive tools which will allow the selection and transfer of a single embryo in each IVF cycle. We propose a pilot study to characterize the metabolic profile of implantable embryos from conditioned culture media using proton nuclear magnetic resonance spectroscopy (1H-NMR).