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: 10515327
• 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/10515327
• 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; Coculture Techniques; Combination Drug Therapy; Data; Deglutition; Diagnosis; Disease; Dysplasia; Electrons; Eosinophilic Infiltrate; Eotaxin; Epithelial Cells; Epithelium; 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; Invaded; 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; Radiation; 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.

低存活率、高吸烟率和高饮酒率以及晚期食管炎的诊断 退伍军人中的鳞状细胞癌(ESCC)需要进一步研究新的治疗方法 鳞癌的治疗策略。在过去的几年里，新兴的免疫治疗方法，如 作为采用细胞转移，已经强调了了解和操纵肿瘤的重要性 微环境。食管鳞癌的肿瘤微环境研究较少。在这份提案中，我们重点关注 确定嗜酸性粒细胞在ESCC中的作用，嗜酸性粒细胞是微环境的关键组成部分。我们已经产生了 初步数据显示在人食管癌肿瘤中存在嗜酸性粒细胞，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)的AS小鼠模型。 确定嗜酸性粒细胞在肿瘤微环境中功能的可行模型。我们表明，在 4-NQO模型中，嗜酸性粒细胞不断地由骨髓产生，并专门招募到 食道异型增生与人类食管鳞癌相似，在无异型增生的区域中不存在。此外，嗜酸性粒细胞趋化因子-1 CCL11是一种有效的嗜酸性粒细胞趋化物质，在异型增生的区域被诱导。这些观察结果表明 嗜酸性粒细胞发挥肿瘤特异性作用，并不是更普遍的炎症反应的结果。而当 4-NQO模型将用于产生对嗜酸性粒细胞在肿瘤发生中的作用的更广泛的理解， 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械性见解 肿瘤生物学中的激活和功能。在这项建议中，我们展示了嗜酸性粒细胞与食道共培养 细胞增加了细胞的凋亡率，降低了存活率，改变了上皮细胞的细胞因子谱，增加了 肿瘤坏死因子-α和IL-2、IL-8、IL-10的表达。此外，产生活性氧的嗜酸性粒细胞 (ROS)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。这一点意义重大 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，导致释放 细胞毒性活性氧化剂的种类。综上所述，这些数据表明嗜酸性粒细胞可能会降低存活率。 ESCC细胞通过释放ROS。谷胱甘肽过氧化物酶1(GPX1)是一种表达丰富的抗氧化酶 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。在这里我们 独立提出GPX1调节嗜酸性粒细胞产生ROS，并减少其表达 嗜酸性粒细胞与食道上皮细胞共培养后检测到的GPX1蛋白被嗜酸性粒细胞修饰。 上皮性相互作用。根据这些数据和嗜酸性粒细胞在不典型增生区域的招募情况 和ESCC中的癌，我们的主要假设是嗜酸性粒细胞对肿瘤细胞和 因此在ESCC中是保护性的。这个项目的具体目标是：1.确定嗜酸性粒细胞的作用 在食管癌发生发展中的作用。我们将利用食管鳞癌的4-NQO模型，比较 CCL11-/-与WT小鼠经4-NQO处理后比较。然后我们将确定嗜酸性粒细胞耗尽的影响 4-NQO处理WT小鼠不同时间点的高嗜酸性粒细胞数。2.界定 嗜酸性粒细胞对上皮信号和动态平衡的影响。我们将测定嗜酸性粒细胞对正常人的影响 和发育不良的上皮细胞，通过与小鼠器官共培养嗜酸性粒细胞。更具体地说，我们将定义 GPX1与野生型嗜酸性粒细胞共培养对嗜酸性粒细胞功能和活化的影响 有机体和类肿瘤。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。