Quantitative assessment of pre-metastatic immune subversion as a risk factor for melanoma relapse

项目总结/摘要 在理解实体瘤首先发生的免疫学悖论方面存在根本性的差距。 通过淋巴管转移到免疫丰富的淋巴结。在此过程中，免疫 监测功能被禁用，最终允许转移性传播到它应该在的精确位置 首先清除肿瘤引流的前哨淋巴结（SLN）。虽然具有明显的预后意义，但很明显， 单独的SLN转移状态不足以确定患者的风险或复发。因此，我们认为， 了解这种慢性肿瘤介导的区域性免疫反应的机制， 免疫抑制，通常称为SLN中的转移前小生境（PMN）形成， 对开发改进的预后和治疗工具以检测和逆转癌症的重要见解 在转移性进展的自然史的早期传播。我们的长期目标是发展 治疗策略能够克服SLN PMN的免疫妥协，从而破坏 癌症转移的第一阶段因此，当前工作的目标是从机制上 直接询问原发性肿瘤淋巴流出物的亚细胞成分的过程 介导PMN形成。中心假说提出，在实体瘤中，亚细胞介质来源于 来自原发性肿瘤微环境的肿瘤细胞通过血管系统活跃地运输， 以在肿瘤引流SLN中产生PMN的方式。这一假设是根据以下事实提出的： 申请人实验室产生的初步数据;即发现和表征 人淋巴细胞外囊泡（L-EV），其具有经证实的调节 免疫功能基本原理断言，在阐明定义PMN的因素和特征时， 在SLN中形成，获得的知识将是重要的，因为它将识别组织病理学生物标志物， 可以帮助患者在SLN中存在黑色素瘤细胞之外进行风险分层。以强为导 初步数据，这一假设将在两个特定的目标进行测试：1）确定机制，淋巴 亚细胞因子促进转移前SLN中的免疫功能障碍， S100 A9）; 2）评估这些免疫调节因子在预测复发风险中的预后效用 通过考虑全面的，互动的细胞景观，定义了免疫学 SLN受损该方法是创新的，因为它将使用机制驱动的模型来识别亚细胞 因子从以前未表征的生物流体，人淋巴（目标1），补充了一种新的， 多重生物标志物成像方法，以便以定量方式调查SLN免疫学景观， 并以空间保留的方式最终将预测特征转化为临床适用的平台 (Aim 2）。这些发现将导致对有复发风险和需要治疗的早期患者的精确定义。 早期干预。

PROJECT SUMMARY/ABSTRACT There is a fundamental gap in understanding the immunological paradox by which solid tumors first metastasize via lymphatic channels into the immune-rich lymph node. During this process, immune surveillance is disabled, ultimately allowing metastatic dissemination to the precise location where it should be first eliminated—the tumor-draining, sentinel lymph node (SLN). While notably prognostic, it is clear the metastatic status of the SLN alone is not sufficient in determining patient’s risk or relapse. Therefore, understanding the mechanisms underlining this chronic process of tumor mediated regional immunosuppression, commonly referred to as pre-metastatic niche (PMN) formation in the SLN will lend significant insights for developing improved prognostic and therapeutic tools to detect and reverse cancer dissemination early in the natural history of metastatic progression. Our long term goal is to develop therapeutic strategies capable of overcoming the immune compromise of the SLN PMN and thereby disrupt the first stage of cancer metastasis. Therefore, the objective of the current work is to mechanistically interrogate the process by which the subcellular component of the primary tumor lymphatic effluent directly mediates PMN formation. The central hypothesis proposes that in solid tumors, subcellular mediators derived from the primary tumor microenvironment actively traffic through the lymphatics and in a cargo-dependent manner create a PMN in the tumor-draining SLN. This hypothesis has been formulated on the basis of preliminary data produced in the applicant’s laboratory; namely the discovery and characterization of human lymphatic extracellular vesicles (L-EV) which have a demonstrated capacity to modulate immune function. The rationale asserts that in elucidating the factors and signatures that define PMN formation in the SLN, the knowledge gained will be significant as it will identify histopathologic biomarkers that could aid in patient risk stratification beyond the presence of melanoma cells in the SLN. Guided by strong preliminary data, this hypothesis will be tested in two specific aims: 1) identify mechanisms whereby lymphatic subcellular factors promote immune dysfunction in the pre-metastatic SLN beyond those already identified (i.e. S100A9); 2) evaluate the prognostic utility of these immune-modulating factors in predicting risk of recurrence by considering the comprehensive, interactive cellular landscape that defines the immunologically compromised SLN. The approach is innovative as it will use a mechanism-driven model to identify subcellular factors from a previously uncharacterized biological fluid, human lymph (Aim 1), complemented by a novel, multiplexed biomarker imaging approach in order to survey the SLN immunological landscape in a quantitative and spatially preserved manner to ultimately translate predictive features into clinically amenable platforms (Aim 2). Such findings will result in a refined definition of early-stage patients at risk of relapse and in need of earlier interventions.