Do Tumor-Immune Interactions Prime Systemic Tolerance of Triple-Negative Breast Cancer Brain Metastases?

项目摘要/摘要 据广泛估计，90%的癌症相关死亡是由转移引起的。这一统计数据突显了我们的无能 一旦癌症在体内扩散，我们就需要更好地了解分子 驱动转移的机制。三阴性乳腺癌是乳腺癌的一种亚型，发生于 10%-20%的病例。作为一个群体，TNBCs在许多方面构成了一个未被满足的临床挑战：(1)TNBCs代表 最具侵袭性和最具转移性的乳腺癌亚型，(2)高达46%的晚期TNBC患者将发展为 脑转移，(3)TNBC患者发生脑表面弥漫性转移的风险是对照组的四倍 脑(软脑膜疾病)，这是迅速和普遍致命的，4)非洲血统的妇女有高达 发展为TNBC的可能性高出80%-使TNBC成为癌症差异。此前的研究表明， 原发TNBC具有高度的免疫原性，免疫浸润与改善预后有关。然而，几乎没有什么是 已知TNBC脑转移的免疫环境以及肿瘤-免疫相互作用的影响 转移潜能。认识到这些紧迫的问题，我选择了把我的职业生涯作为一个独立的癌症 研究人员使用系统生物学方法来揭示转移和种族背后的分子机制- 具体的癌症差异。将执行全面的指导、研究和职业发展计划 在K99和R00培训期间，这将为我提供必要的工具来 向独立的早期过渡。首先，为了可视化肿瘤与免疫的相互作用，我将构建一张原位蛋白质图 使用MIBI的TNBC脑转移--一种尖端的多路成像方法(AIM 1)。我将使用经过验证的成像 分析管道以量化肿瘤免疫微环境的组成和空间结构以及 确定这些特征与患者预后的相关性程度。在AIM 2中，我将识别肿瘤免疫 用单细胞RNA测序技术对TNBC脑转移瘤样本进行受体-配体对。最后，在AIM 3中，我将使用 MIBI可视化原发TNBCs中的肿瘤-免疫相互作用以确定原发肿瘤中的相互作用 微环境导致脑转移的播散性肿瘤细胞对免疫系统的耐受性。我会验证的 通过检测相关靶点在患者脑脊液(CSF)中的表达，脑脊液中含有脑肿瘤- 相关的无细胞RNA。我提议的博士后研究结果将对公共卫生产生积极影响，因为 他们将揭示负责启动免疫系统以防止转移的关键肿瘤-免疫相互作用，以及 将产生首个“TNBC脑转移相互作用体”。我的结果将导致新分子的发现 这些目标的首要目标是降低因转移而导致的癌症死亡率。K99职业发展计划和 有重点的研究培训对于扩展我在生物计算方面的技能至关重要--生物计算是我 建议的研究；以及填补我背景中的任何空白，为我及时和成功地过渡到 R00独立阶段。总体而言，双阶段奖将显著加强我的研究，并确保我的 作为一名公认的癌症研究人员，继续为公共卫生做出贡献，并增加科学的多样性。

PROJECT ABSTRACT/SUMMARY It is widely estimated that 90% of cancer-related deaths are caused by metastasis. This statistic underscores our inability to manage cancer once it disseminates through the body, and our need to better understand the molecular mechanisms that drive metastasis. Triple-negative breast cancer (TNBC) is a subtype of breast cancer that occurs in 10-20% of cases. Taken as a group, TNBCs pose an unmet clinical challenge in many ways: (1) TNBCs represents the most aggressive and most metastatic subtype of breast cancer, (2) up to 46% of late-stage TNBC patients will develop brain metastases, (3) TNBC patients are at four-times higher risk of developing diffuse metastases on the surface of the brain (leptomeningeal disease), which is rapidly and universally fatal, and 4) women of African ancestry have an up to 80% higher likelihood of developing TNBC – making TNBC a cancer disparity. Previous studies demonstrate that primary TNBC is highly immunogenic, and immune infiltration is associated with improved prognosis. However, little is known about the immune environment in TNBC brain metastases and how tumor-immune interactions effect metastatic potential. Recognizing these pressing issues, I have chosen to focus my career as an independent cancer researcher on using a systems biology approach to uncover molecular mechanisms that underlie metastasis and race- specific cancer disparities. A comprehensive mentoring, research, and career development plan will be executed over the course of the K99 and R00 training period, which will provide me with the necessary tools to make an early transition to independence. First, to visualize tumor-immune interactions, I will construct an in-situ protein map of TNBC brain metastases using MIBI – a cutting-edge multiplexed imaging method (AIM 1). I will use a validated imaging analysis pipeline to quantitate the composition and spatial architecture of the tumor-immune microenvironment and determine the extent to which these features correlate with patient outcomes. In AIM 2, I will identify tumor-immune receptor-ligand pairs using single-cell RNA-sequencing on TNBC brain metastases samples. Lastly, in AIM 3, I will use MIBI to visualize tumor-immune interactions in primary TNBCs to determine whether interactions in the primary tumor microenvironment prime immune system tolerance of disseminated tumor cells enabling brain metastases. I will validate relevant targets by measuring their expression in patient cerebral spinal fluid (CSF), which contains brain-tumor- associated cell-free RNA. The results of my proposed postdoctoral research will positively impact public health as they will reveal key tumor-immune interactions responsible for priming the immune system for metastasis, and will generate the first “TNBC brain metastasis interactome”. My results will lead to the discovery of new molecular targets with the primary goal of reducing metastasis-driven cancer mortality. The K99 career development plan and focused research training will be critical to expand my skillset in biocomputation – a necessary component of my proposed research; as well as fill any gaps in my background, preparing me for a timely and successful transition to the R00 independent phase. Overall, the dual-phase award will significantly enhance my research, and ensure my continued contribution to public health and increasing diversity in science, as an established cancer researcher.