Assessment of microvessel density and tumor proximity for prognosis of cancer

描述(申请人提供)：肿瘤微环境(TME)对肿瘤进展、复发和转移有重要作用。目前，还没有一种标准化的方法来同时定量TME的主要成分，如淋巴血管侵犯，以及预测肿瘤的局部复发和转移。这个项目的主要目标是开发一种利用基于量子点(QD)的免疫组织化学荧光(IHF)来量化TME签名的技术。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，包括局部区域复发和转移的风险。我们最近开发了基于量子点的IHF技术，用于多重生物标志物的量化。我们的初步研究支持我们完成这项研究的目标，即开发一种分子成像技术来量化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。预计使用这项技术，HNC病理学家将能够更准确地识别预后不良的高危患者。这将为头颈部癌症外科医生、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。这项技术还有可能在适当的修改后应用于其他类型的癌症的治疗和护理。该项目将涉及2个具体目标和5个可衡量的里程碑。目的：制备和验证特异性识别血管和淋巴管的QD抗体结合物，以及福尔马林固定石蜡包埋(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结合物；(2)建立TME标记密度及其到肿瘤巢的距离的定量方法；(3)通过与传统IHC的比较，验证基于QD的IHF的有效性。目的：探讨微血管密度和肿瘤近似性是否可以作为判断预后的生物标志物，并探讨其与喉癌局部复发和淋巴结转移的关系。这一目标的里程碑是(4)使用来自有或没有淋巴结转移和复发疾病的HNC患者的100个FFPE样本完成TME信号的染色和量化；(5)完成将微血管密度和肿瘤邻近程度与HNC患者的临床特征相关联的统计分析。

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.