(PQ3) Cellular and Molecular Mechanisms Driving Myeloid Compartment Variation in Human Triple Negative Breast Cancer

项目总结 在过去的十年里，许多研究检查了癌症患者的肿瘤和血液隔间。这些 研究增加了我们对癌症免疫变化的了解，但未能完全解释 髓系抗原提呈细胞(MAPC)的功能变异。我们必须先解决这一差距，然后才能 开始全面回答上面提出的问题，因为MAPC控制向T细胞递送癌症抗原 从而启动和调节抗癌免疫。这项提议的首要目标是更好地 了解三阴性乳腺癌(TNBC)免疫反应变异的分子基础 单细胞水平。这种方法可能会重新定义癌症中MAPC的生物学。我们提出了一种系统的 75例TNBC残留癌患者血液和肿瘤免疫状态的研究我们 假设TNBC患者MAPC的转录组和功能变化与特定的 其相关癌细胞的基因组图谱，并有助于对化疗和 疾病的发展。我们提出了两个具体目标：在目标1中，我们将应用单细胞转录技术来 鉴定血液中髓系APC特异性转录图谱。我们将在稳定状态下这样做，作为回应 肿瘤衍生因子的体外激活。这将识别不同的转录和功能模块 这可以预测肿瘤环境改变时的细胞行为。基因集和/或细胞 在血液研究中定义的将应用于肿瘤，以使用基于质谱学的成像来追踪这些细胞 并检查宏观解剖收获的白细胞浸润物的特异性转录图谱。在目标2中， 我们将检查可能与肿瘤中APC的吸引和分化有关的基因。 (趋化因子和细胞因子)。我们将研究癌症基因组异常和转录之间的联系。 APC的变化以及髓系APC对肿瘤产物的体外反应和原位反应 转录档案。我们将检查TNBC患者来源的异种移植组织 残留病变确定癌细胞内在趋化因子/细胞因子谱及其在吸引肿瘤细胞中的作用 将髓系细胞植入人源化小鼠体内。CRISPR/Cas9编辑将应用于癌细胞以 产生变异并验证与APC隔室中的变异和对肿瘤的影响之间的联系 进展和转移。这些研究将为更好地理解免疫 变化有助于TNBC的进取性，并使识别潜在的可操作 残留病的治疗靶点。

PROJECT SUMMARY Numerous studies over the past decade examined the tumor and blood compartment in cancer patients. These studies have added to our understanding of immune alterations in cancer, but have failed to fully explain the functional variation in myeloid antigen presenting cells (mAPCs). We must address this gap before we can begin to fully answer the question posed above because mAPCs control cancer antigen presentation to T cells thereby launching and regulating anti-cancer immunity. The overarching goal of this proposal is to better understand the molecular basis of variation in the immune response to triple negative breast cancer (TNBC) at single cell level. This approach will likely redefine the biology of mAPCs in cancer. We propose a systematic exploration of immune status in blood and tumors in a cohort of 75 TNBC patients with residual cancer. We hypothesize that the variation in transcriptome and function of mAPCs in TNBC patients is linked with specific genomic profiles of their associated cancer cells and contributes to variable response to chemotherapy and disease progression. We propose two specific aims: In Aim 1, we will apply single cell transcriptomics to identify myeloid APC-specific transcriptional profiles in the blood. We will do so at steady state and in response to ex vivo activation with tumor derived factors. This will identify distinct transcriptomics and functional modules that could enable prediction of cell behavior when tumor environments are altered. Gene sets and/or cells defined in blood studies will be applied to tumors to trace these cells using mass spectrometry-based imaging and to examine transcriptional profiles specific to leukocyte infiltrate harvested with macro dissection. In Aim 2, we will examine genes that could be linked with the attraction and differentiation of APCs in the tumor (chemokines and cytokines). We will examine links between cancer genomic aberrations and transcriptional variation in APCs and between the ex vivo response of myeloid APCs to tumor products and in situ transcriptional profiles. We will examine TNBC patient derived xenograft (PDX) tissues established from residual disease to determine cancer-cell intrinsic chemokine/cytokine profiles, and their role in the attraction of myeloid cells when implanted in humanized mice. CRISPR/Cas9 editing will be applied to cancer cells to generate variants and to verify links with the variation in the APCs compartment and impact on tumor progression and metastasis. These studies will lay a foundation for better understanding how immune variations contribute to the aggressiveness of TNBC and enable the identification of potential actionable therapeutic targets in residual disease.