Harnessing the nervous system to overcome resistance to immunotherapy in oral cancer

项目总结/摘要 为了使其生长和转移潜力最大化，实体瘤促进肿瘤内新神经纤维的形成 微环境（TME）。在患有口腔癌、前列腺癌、乳腺癌、胃癌、胰腺癌和其他癌症的患者中， TME中的神经纤维与不良的临床结果相关。我们证明了口腔癌细胞诱导了一种独特的 TME中肿瘤相关神经元（TAN）的异质性组成。神经系统扮演重要角色 在体内平衡和组织炎症反应中的作用。然而，神经对免疫细胞的调节在很大程度上仍然存在。 不清楚我们的长期目标是阐明推动癌症进展的相互神经癌信号，以确定 用于治疗和克服免疫疗法抗性的新靶标。我们的初步数据显示， 通过免疫调节分子的表达直接与免疫细胞沟通， 旁分泌，肾上腺素能依赖性癌细胞信号传导。我们将在拟议项目中测试的总体假设是 TAN诱导了支持肿瘤进展的适应不良免疫应答。这些新形成的，重新编程的 TAN通过多步骤机制调节免疫反应，包括静止神经元的转化 进入发芽细胞，可以渗透并与其他细胞类型相互作用，释放肾上腺素能神经活性分子， 支持免疫抑制微环境的发展。这些步骤中的每一个都可能促进肿瘤 进展和治疗抵抗。拟议的研究是创新的，因为它将利用新的概念， 免疫学和癌症生物学使用先进的模型系统，以深入了解肿瘤的机制 研究进展并基于神经免疫串扰识别癌症治疗的新靶点。这种跨学科 该提案将联合收割机肿瘤学、免疫学、细胞生物学、神经生物学、癌症遗传学、病理学和 在两个实验室（Amit和Calin）的两个特定目标中进行生物统计。目标1：确定 神经元依赖性癌细胞信号传导调节细胞毒性T细胞功能。我们将使用药理学和遗传学 在同基因小鼠模型中结合先进的空间成像技术（蛋白质和RNA）的方法 了解重编程的神经元如何调节细胞毒性T细胞的抗肿瘤活性。解读TAN如何发挥两者的作用 通过肾上腺素能信号传导和免疫检查点表达的抗肿瘤免疫激活和抑制活性 将使我们能够利用安全，负担得起和完善的神经调节方法来克服 癌症中的免疫抑制。目的2：鉴定TAN的胞外囊泡穿梭驱动miRNAs 重编程及其在口腔癌进展中的作用。使用人类来源的感觉神经元和功能 通过基因组方法，我们将研究TAN中免疫调节基因的miRNA依赖性功能可塑性。 完成拟议的研究将为针对神经机制的治疗策略铺平道路 与免疫抑制和对免疫疗法的逆转抗性相关。针对此的治疗方法 预期肿瘤生物学的关键组成部分可以改善患者的生存率、治疗反应和生活质量。

PROJECT SUMMARY/ABSTRACT To maximize their growth and metastatic potential, solid tumors promote the formation of new nerve fibers in the tumor microenvironment (TME). In patients with oral, prostate, breast, gastric, pancreatic, and other cancers, high densities of nerve fibers in the TME are associated with poor clinical outcomes. We proved that oral cancer cells induce a unique heterogeneous composition of tumor-associated neurons (TANs) in the TME. The nervous system plays important roles in homeostasis and inflammatory responses in tissues. However, the regulation of immune cells by nerves remains largely unclear. Our long-term goal is to elucidate the reciprocal nerve-cancer signals that drive cancer progression to identify novel targets for therapy and for overcoming immunotherapy resistance. Our preliminary data show that neurons communicate with immune cells directly through the expression of immunomodulatory molecules and indirectly through paracrine, adrenergic-dependent cancer cell signaling. The overall hypothesis that we will test in the proposed project is that TANs induce a maladaptive immune response that supports tumor progression. These newly formed, reprogrammed TANs regulate the immune response through a multistep mechanism that includes the transformation of quiescent neurons into sprouting cells that can infiltrate and interact with other cell types, release adrenergic neuroactive molecules, and support the development of an immunosuppressive microenvironment. Each of these steps may promote tumor progression and therapy resistance. The proposed research is innovative because it will capitalize on new concepts in immunology and cancer biology using advanced model systems to yield insights into the mechanisms of tumor progression and identify new targets for cancer therapy based on neuro-immune crosstalk. This cross-disciplinary proposal will combine expertise from oncology, immunology, cell biology, neurobiology, cancer genetics, pathology, and biostatistics in two specific aims across the two labs (Amit and Calin). Aim 1: Determine the mechanisms by which neuron-dependent cancer cell signaling regulates cytotoxic T-cell function. We will use pharmacological and genetic approaches combined with advanced spatial imaging techniques (for both protein and RNA) in syngeneic mouse models to understand how reprogrammed neurons regulate cytotoxic T-cell antitumor activity. Deciphering how TANs exert both antitumor immune activation and suppression activity through adrenergic signaling and immune checkpoint expression respectively, will allow us to leverage safe, affordable and well established neuromodulatory approaches to overcome immunosuppression in cancer. Aim 2: Identify the extracellular vesicle-shuttled driver miRNAs of TAN reprogramming and their roles in oral cancer progression. Using human-derived sensory neurons and functional genomic approaches, we will investigate the miRNA-dependent functional plasticity of immunomodulatory genes in TANs. The completion of the proposed studies will pave the way for treatment strategies that target the neuronal mechanisms associated with immunosuppression and reverse resistance to immunotherapy. Therapeutic approaches targeting this critical component of tumor biology are anticipated to improve patients' survival, treatment responses, and quality of life.