The role of eosinophils and oxidative stress in esophageal malignancy

嗜酸性粒细胞和氧化应激在食管恶性肿瘤中的作用

• 批准号: 10292942
• 负责人: Yash Choksi
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-10-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292942
• 关键词: 3-Dimensional; Adoptive Cell Transfers; Aftercare; Alcohol consumption; Apoptosis; Area; Biology; Blood Vessels; Bone Marrow; CCL11 gene; Carcinoma; Cells; Cellular Structures; Chemotactic Factors; Chemotherapy and/or radiation; Coculture Techniques; Combination Drug Therapy; Culture Techniques; Data; Deglutition; Diagnosis; Disease; Dysplasia; Electrons; Eosinophilia; Eotaxin; Epithelial; Epithelial Cells; Equilibrium; Esophageal Intraepithelial Neoplasia; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Esophageal carcinoma; Esophagectomy; Esophagus; High Prevalence; Homeostasis; Human; Hydrogen Peroxide; IL8 gene; Immune; Immune checkpoint inhibitor; Immunotherapy; Infiltration; Inflammatory Response; Interleukin-10; Interleukin-2; Interleukin-5; Lead; Light; Lipids; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of esophagus; Mediating; Metastatic Neoplasm to Lymph Nodes; Methods; Modeling; Modification; Mus; Operative Surgical Procedures; Organoids; Oxidants; Oxidative Stress; Oxides; Pathway interactions; Patients; Peroxidases; Play; Population; Production; Reactive Oxygen Species; Reduced Glutathione; Regulation; Role; Signal Transduction; Squamous cell carcinoma; Stage at Diagnosis; TNF gene; Testing; Therapeutic Intervention; Thiocyanates; Time; Tissues; Tumor Biology; Tumor Immunity; Water; advanced disease; antioxidant enzyme; cellular targeting; clinically relevant; cytokine; cytotoxic; eosinophil; eosinophil peroxidase; esophageal squamous cell cancer; glutathione peroxidase; health related quality of life; human disease; improved; insight; mouse model; neoplasm registry; neoplastic cell; oxidation; recruit; smoking prevalence; tumor; tumor initiation; tumor microenvironment; tumor progression; tumorigenesis; tumorigenic

The poor survival, high prevalence of smoking and alcohol use, and advanced stage at diagnosis of esophageal squamous cell carcinoma (ESCC) in the VA population necessitates further study into new management strategies for squamous cell carcinoma. Over the past few years, emerging immunotherapy approaches, such as adoptive cell transfer, have highlighted the importance of understanding and manipulating the tumor microenvironment. The tumor microenvironment in ESCC is understudied. In this proposal, we focus on determining the role of eosinophils, a key component of the microenvironment, in ESCC. We have generated preliminary data demonstrating the presence of eosinophils in human ESCC tumors but not in tumor-adjacent tissue. We then established and characterized the 4-nitroquinolone-1-oxide (4-NQO) murine model of as a feasible model for determining the function of eosinophils in the tumor microenvironment. We show that, in the 4-NQO model, eosinophils are constantly being produced by the bone marrow, recruited specifically to areas of esophageal dysplasia as in human ESCC, and not present in areas without dysplasia. Additionally, Eotaxin-1 (CCL11), a potent eosinophil chemoattractant, is induced in areas of dysplasia. These observations suggest that eosinophils play a tumor specific role and are not the result of a more general inflammatory response. While the 4-NQO model will be employed to generate a broader understanding of eosinophils’ role in tumorigenesis, sophisticated co-culture techniques the we have developed provide mechanistic insights into eosinophil activation and function in tumor biology. In this proposal we show that eosinophils co-cultured with esophageal cells increases apoptosis, decreases viability, and alters the cytokine profile of epithelial cells, increasing expression of TNF-a and IL-2, IL-8, and IL-10. Furthermore, eosinophils, which produce reactive oxygen species (ROS), have increased expression of eosinophil peroxidase after co-culture with epithelial cells. This is significant as eosinophil peroxidase is known to catalyze the oxidation of halides and thiocyanate, resulting in the release of cytotoxic reactive oxidant species. Taken together, these data suggest that eosinophils may decrease viability ESCC cells via release of ROS. Glutathione Peroxidase 1 (Gpx1), an abundant antioxidant enzyme expressed by eosinophils, catalyzes the reduction of hydrogen peroxide or lipid peroxidases to water. Here we independently propose that Gpx1 regulates ROS production by eosinophils, and the decrease in expression of Gpx1 by eosinophils after co-culture with esophageal epithelial cells suggests it is modified by the eosinophil- epithelial interaction. In light of these data and the recruitment of eosinophils specifically to areas of dysplasia and carcinoma in ESCC, our overarching hypothesis is that eosinophils are cytotoxic to tumor cells and thus are protective in ESCC. The specific aims for this project are: 1. To determine the role of eosinophils in ESCC tumorigenesis. We will utilize the 4-NQO model of ESCC and compare esophageal tumorigenesis in Ccl11-/- versus WT mice after treatment with 4-NQO. We will then determine the impact of eosinophil depletion and hyper-eosinophilia on WT mice treated with 4-NQO at different time points. 2. To define the effect of eosinophils on epithelial signaling and homeostasis. We will determine the effect of eosinophils on normal and dysplastic epithelium by co-culturing eosinophils with mouse organoids. More specifically, we will define the role of Gpx1 on eosinophil function and activation by co-culturing Gpx1-/- eosinophils with both wild-type organoids and tumoroids. These studies have the potential to improve understanding of eosinophil biology within the context of ESCC, which could lead to specific therapeutic interventions targeting eosinophil function.

生存率低，吸烟和饮酒的高流行率，以及诊断食管癌时的晚期， VA人群中的鳞状细胞癌（ESCC）需要进一步研究新的治疗方法 鳞状细胞癌的治疗策略在过去的几年里，新兴的免疫治疗方法，如 作为过继性细胞转移，强调了理解和操纵肿瘤的重要性。 微环境ESCC中的肿瘤微环境研究不足。在本提案中，我们重点关注 确定ESCC中嗜酸性粒细胞（微环境的关键成分）的作用。我们已经生成 初步数据表明，嗜酸性粒细胞存在于人食管鳞癌肿瘤中，但在癌旁组织中不存在。 组织.然后，我们建立并表征了4-硝基喹诺酮-1-氧化物（4-NQO）小鼠模型， 确定肿瘤微环境中嗜酸性粒细胞功能的可行模型。我们表明，在 4-在NQO模型中，嗜酸性粒细胞不断由骨髓产生，特异性地募集到NQO区域。 食管发育不良，如人ESCC中的食管发育不良，并且不存在于没有发育不良的区域。此外，Eotaxin-1 （CCL 11），一种有效的嗜酸性粒细胞化学引诱物，在发育不良的区域被诱导。这些观察提示 嗜酸性粒细胞发挥肿瘤特异性作用，并且不是更普遍的炎症反应的结果。而 4-NQO模型将用于更广泛地理解嗜酸性粒细胞在肿瘤发生中的作用， 我们开发的复杂的共培养技术提供了对嗜酸性粒细胞的机制性见解， 在肿瘤生物学中的激活和功能。在这个建议中，我们表明嗜酸性粒细胞与食管癌细胞共培养， 细胞凋亡增加，活力降低，改变上皮细胞的细胞因子谱， TNF-α和IL-2、IL-8和IL-10的表达。此外，产生活性氧的嗜酸性粒细胞 (ROS)在与上皮细胞共培养后，嗜酸性粒细胞过氧化物酶的表达增加。这是重要 因为已知嗜酸性粒细胞过氧化物酶催化卤化物和硫氰酸盐的氧化，导致 细胞毒性活性氧化剂种类。综上所述，这些数据表明，嗜酸性粒细胞可能会降低生存能力， ESCC细胞通过释放ROS。谷胱甘肽过氧化物酶1（Gpx 1）是一种在大肠杆菌中表达的丰富的抗氧化酶， 通过嗜酸性粒细胞，催化过氧化氢或脂质过氧化物酶还原为水。这里我们 独立提出Gpx 1调节嗜酸性粒细胞产生ROS， 与食管上皮细胞共培养后嗜酸性粒细胞的gpx 1表明它被嗜酸性粒细胞修饰， 上皮相互作用根据这些数据以及嗜酸性粒细胞在异型增生区域的募集 和癌，我们的总体假设是嗜酸性粒细胞对肿瘤细胞具有细胞毒性， 因此在ESCC中是保护性的。该项目的具体目标是：1。为了确定嗜酸性粒细胞的作用 在ESCC肿瘤发生中的作用我们将利用ESCC的4-NQO模型，比较 4-NQO处理后Ccl 11-/-vs WT小鼠。然后我们将确定嗜酸性粒细胞减少的影响 和在不同时间点用4-NQO处理的WT小鼠的高嗜酸性粒细胞增多。2.定义的效果 嗜酸性粒细胞对上皮信号传导和稳态的影响。我们将确定嗜酸性粒细胞对正常 和发育不良的上皮细胞。更具体地说，我们将定义 通过Gpx 1-/-嗜酸性粒细胞与两种野生型共培养，Gpx 1对嗜酸性粒细胞功能和活化的作用 类器官和类肿瘤。这些研究有可能提高对嗜酸性粒细胞生物学的理解， ESCC的背景，这可能导致针对嗜酸性粒细胞功能的特定治疗干预。