Development of lavage EV protein biomarkers for minimally-invasive detection of endometrial cancer

项目总结 早期发现子宫内膜癌可以将5年生存率从20%提高到95%。 然而，目前还没有非侵入性的检测方法。我们建议使用 细胞分泌的胞外小泡(EVS)作为子宫新诊断标志物的来源 灌洗液--一种更针对子宫内膜癌的样本。在生物标志物的范围内 检测，电动汽车为临床分析提供了许多好处，包括非侵入性收集， 适合纵向疾病监测的样品来源，较高的稳定性和样本量， 处理时间更快，成本更低。然而，对exosome和其他EV的分析还没有 真的有可能及早发现子宫内膜癌。生物液EV的常见操作方法 生物标志物分析非常长和繁琐，因为目标水平极低，而目标水平很高 游离蛋白质的背景。为了更好地发现生物标记物和子宫内膜癌 诊断时，需要一种更可靠、更有效的方法，能够丰富潜力 具有更高纯度的癌症相关蛋白。在NIH SBIR第一阶段研究中，我们将实施我们的 一种快速、重现性好、回收率高达95%、99.9%的EVS富集法 从灌洗液样本中分析子宫内膜癌的纯度。我们已经进行了初步的 Wavage EV验证和生物标记物发现实验，并选择了十几个 能够有效区分子宫内膜癌和子宫内膜癌的高质量蛋白质靶点 非癌症对照组。以下目标将在提案的第一阶段完成：目标1： 确定已发现的子宫内膜癌生物标记物从灌洗液EVS到 辨别疾病，精炼面板。目标2：评估和实施新的子宫 灌洗液收集装置，用于快速有效地隔离和测量EV。通过完成 这个项目，来自Wavage EVS的非侵入性子宫内膜癌生物标记物发现平台将 将开发一种微创早期子宫内膜癌检测方法，并将验证其有效性 这可以克服目前方法的局限性，因此可能会有大量的公众 对健康的影响和市场潜力。

PROJECT SUMMARY Early detection of endometrial cancer can increase the 5-year survival rate from <20% to >95%. However, currently there are no non-invasive methods for its detection. We propose the use of cell-secreted extracellular vesicles (EVs) as the source of new diagnostic biomarkers from uterine lavage – a sample that is much more targeted to endometrial cancer. Within the scope of biomarker detection, EVs offer numerous benefits for clinical analysis, including non-invasive collection, a suitable sample source for longitudinal disease monitoring, higher stability and sample volumes, faster processing times and lower cost. However, the analysis of exosomes and other EV has not really been possible for early endometrial cancer detection. The common procedures for biofluid EV biomarker analysis are very long and cumbersome due to extremely low target levels and high background of free proteins. In order to enable better biomarker discovery and endometrial cancer diagnosis, a more reliable and efficient approach is needed, capable of enriching potential cancer-associated proteins with higher purity. In this NIH SBIR Phase I study, we will implement our novel method for fast and reproducible enrichment of EVs with >95% recovery yield and >99.9% purity for analysis of endometrial cancer from lavage samples. We have already carried out initial lavage EV validation and biomarker discovery experiments, and selected over a dozen of high-quality protein targets that enable effective differentiation of endometrial cancer from non-cancer controls. The following aims will be completed in the Phase I of the proposal: Aim #1: Determine the feasibility of the discovered endometrial cancer biomarkers from lavage EVs to differentiate the disease and refine the panel. Aim #2: Evaluate and implement the novel uterine lavage collection device for fast and effective EV isolation and measurement. By the completion of this project, the non-invasive endometrial cancer biomarker discovery platform from lavage EVs will be developed, and a minimally invasive early endometrial cancer detection assay will be validated that can overcome the limitations of current approaches, and thus could have an enormous public health impact and market potential.