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

项目摘要 早期发现子宫内膜癌可以提高5年生存率从<20% to >95%。 然而，目前还没有非侵入性的检测方法。我们建议使用 细胞分泌的细胞外囊泡（EV）作为子宫内膜癌新诊断生物标志物的来源 灌洗-一种更有针对性的子宫内膜癌样本。在生物标志物范围内 检测，EV为临床分析提供了许多好处，包括非侵入性收集， 用于纵向疾病监测的合适样品源、更高的稳定性和样品体积， 更快的处理时间和更低的成本。然而，对外来体和其他EV的分析还没有 真的有可能用于早期子宫内膜癌的检测。生物流体EV的常见程序 生物标志物分析由于极低的目标水平和高的 游离蛋白质的背景。为了更好地发现生物标志物和子宫内膜癌 诊断，需要一种更可靠和有效的方法，能够丰富潜在的 纯度更高的癌症相关蛋白。在这项NIH SBIR I期研究中，我们将实施我们的 一种快速、可重复富集EV的新方法，回收率&gt;95%，回收率&gt;99.9% 用于分析来自灌洗样品的子宫内膜癌的纯度。我们已经进行了初步的 灌洗EV验证和生物标志物发现实验，并选择了十几个 高质量的蛋白质靶点，能够有效区分子宫内膜癌和 非癌症对照。本提案第一阶段将完成以下目标：目标1： 确定从灌洗EV中发现的子宫内膜癌生物标志物的可行性 区分疾病并完善面板。目的#2：评估和实施新型子宫切除术 用于快速有效EV隔离和测量灌洗收集装置。通过完成 这个项目，来自灌洗EV的非侵入性子宫内膜癌生物标志物发现平台将 将开发一种微创早期子宫内膜癌检测方法， 这可以克服目前方法的局限性，因此可以有一个巨大的公众， 健康影响和市场潜力。

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.