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

项目摘要/总结 据广泛估计，90%的癌症相关死亡是由转移引起的。这一统计数字强调了我们无法 一旦癌症扩散到全身，我们需要更好地了解分子水平， 驱动转移的机制。三阴性乳腺癌（TNBC）是乳腺癌的一种亚型， 10-20%的病例。作为一个组，TNBC在许多方面提出了未满足的临床挑战： 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患者在肿瘤表面发生弥漫性转移的风险高四倍。 脑（软脑膜疾病），这是迅速和普遍致命的，和4）非洲血统的妇女有高达 发展TNBC的可能性增加80%-使TNBC成为癌症差异。先前的研究表明， 原发性TNBC具有高度免疫原性，并且免疫浸润与改善的预后相关。然而， 了解TNBC脑转移的免疫环境以及肿瘤-免疫相互作用如何影响 转移潜能认识到这些紧迫的问题，我选择了专注于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R 00培训期间，这将为我提供必要的工具， 尽早过渡到独立。首先，为了可视化肿瘤-免疫相互作用，我将构建一个原位蛋白质图谱， 使用MIBI的TNBC脑转移瘤-一种尖端的多路复用成像方法（AIM 1）。我会用经过验证的图像 分析管道，以定量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结局相关的程度。在AIM 2中，我将识别肿瘤免疫 使用单细胞RNA测序对TNBC脑转移样品进行受体-配体对。最后，在AIM 3中，我将使用 MIBI以使原发性TNBC中的肿瘤-免疫相互作用可视化，以确定原发性肿瘤中的相互作用是否 微环境引发免疫系统对播散的肿瘤细胞的耐受，从而使脑转移成为可能。我会验证 通过测量它们在患者脑脊液（CSF）中的表达来检测相关靶点，所述脑脊液含有脑肿瘤， 相关的无细胞RNA。我提出的博士后研究结果将对公共卫生产生积极影响， 它们将揭示负责启动免疫系统进行转移的关键肿瘤免疫相互作用， 将产生第一个“TNBC脑转移相互作用组”。我的研究结果将有助于发现 目标是降低转移驱动的癌症死亡率。K99职业发展计划和 集中的研究培训将是至关重要的，以扩大我的技能在生物计算-一个必要的组成部分，我的 建议的研究;以及填补我的背景中的任何空白，为我及时成功地过渡到 R 00独立阶段。总的来说，双阶段奖将大大提高我的研究，并确保我的 作为一名知名的癌症研究人员，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.