Supplementary for Assessment of Microvessel Density and Tumor Proximity for Prognosis of Cancer

描述（由申请人提供）：肿瘤微环境（Tumor microenvironment， TME）对肿瘤的进展、复发和转移起着至关重要的作用。目前，还没有标准化的方法来同时量化TME的主要成分，如淋巴血管侵袭，并预测癌症的局部复发和转移。本项目的主要目标是利用基于量子点（QD）的免疫组织荧光（IHF）技术开发一种量化TME特征的技术。我们期望TME成分的单一或多重生物标志物可用于评估预后，包括局部复发和转移的风险。我们最近开发了基于量子点的IHF技术，用于定量多种生物标志物。我们的初步研究支持我们完成本研究的目的，即开发一种分子成像技术，用于量化TME的主要特征，作为评估预后和预测复发和转移的工具。在这个项目中，我们将专注于头颈癌（HNC）的技术开发。预计使用该技术，HNC病理学家将能够更准确地识别预后不良的高风险患者。这将为头颈癌外科医生、放射肿瘤学家和内科肿瘤学家提供有价值的信息，以做出适当的治疗决定。这项技术也有潜力应用于治疗和护理其他类型的癌症适当修改。该项目将涉及2个具体目标和5个可衡量的里程碑。目的1：制备并验证特异性识别血液和淋巴血管的QD-抗体（Ab）偶联物，以及福尔马林固定和石蜡包埋（FFPE）癌组织中的EGFR。这一目标的里程碑是：(1)完成和优化4个具有高结合亲和力和选择性的QD-Ab偶联物；(2)建立TME信号密度及其与肿瘤巢距离的量化方法；(3)通过与传统免疫组化方法的比较，验证了基于量子点的免疫组化方法。目的2：探讨微血管密度和肿瘤邻近度是否可以作为HNC的预后生物标志物，并将其与局部复发和淋巴结转移联系起来。该目标的里程碑是(4)使用100个来自有或无淋巴结转移和复发性疾病的HNC患者的FFPE样本完成TME特征的染色和定量；(5)完成微血管密度、肿瘤邻近度与HNC患者临床特征的相关性统计分析。

DESCRIPTION (provided by applicant): Tumor microenvironment (TME) makes an essential contribution to tumor progression, recurrence, and metastasis. Currently, there is no standardized method to quantify major components of TME simultaneously, such as lymphovascular invasion, and to predict locoregional recurrence and metastasis of cancer. The primary goal of this project is to develop a technology for quantification of the TME signatures by using quantum dot (QD)-based immunohistofluorescence (IHF). We expect that either single or multiplexed biomarkers for the TME components can be used to assess prognosis including risk of the locoregional recurrence and metastasis. We have recently developed QD-based IHF technology for quantification of multiplexed biomarkers. Our preliminary study support us to accomplish the objective of this study, that is, to develop a molecular imaging technology for quantification of major signatures of TME as a tool for assessment of the prognosis and prediction of the recurrence and metastasis. In this project, we will focus on technique development using head and neck cancer (HNC). It is anticipated that using this technology, HNC pathologists will be able to more accurately identify patients who are at high risk for poor prognosis. This would provide valuable information to head and neck cancer surgeons, radiation oncologist, and medical oncologists to make appropriate treatment decisions. This technology also has the potential to be applied to the treatment and care of other types of cancer with appropriate modifications. The project will involve 2 Specific Aims with 5 measurable milestones. Aim 1: To prepare and validate QD- antibody (Ab) conjugates which specifically recognize blood and lymphatic vasculatures, and EGFR in formalin-fixed and paraffin embedded (FFPE) cancer tissues. Milestones for this aim are (1) completion and optimization of 4 QD-Ab conjugates with high binding affinity and selectivity; (2) establishment of the quantification method for both density of the TME signatures and their distance to tumor nests; (3) validation of the QD-based IHF by comparing this method with the conventional IHC. Aim 2: To examine whether microvessel density and tumor proximity can serve as a prognostic biomarkers and correlate them with locoregional recurrence and lymph node metastasis of HNC. Milestones for this aim are (4) completion of staining and quantification of the TME signatures using 100 FFPE samples from HNC patients with and without lymph node metastasis and recurrent disease; (5) completion of statistical analysis to correlate microvessel density and tumor proximity with clinical characteristics of HNC patients.