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、吸烟和饮酒患病率高、食管癌诊断时已处于晚期 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几年中不断出现，例如免疫治疗方法。 作为过继细胞转移，强调了理解和操纵肿瘤的重要性 在本提案中，我们重点研究了 ESCC 中的肿瘤微环境。 确定嗜酸性粒细胞（微环境的关键组成部分）在 ESCC 中的作用。 初步数据表明，人类食管鳞癌肿瘤中存在嗜酸性粒细胞，但肿瘤邻近区域不存在嗜酸性粒细胞 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。 我们证明，在肿瘤微环境中确定嗜酸性粒细胞功能的可行模型。 4-NQO 模型中，骨髓不断产生嗜酸性粒细胞，专门招募到以下区域 食管发育不良与人类食管鳞癌一样，并且不存在于没有发育不良的区域。 (CCL11) 是一种有效的嗜酸性粒细胞趋化剂，在发育异常区域被诱导。 嗜酸性粒细胞发挥肿瘤特异性作用，而不是更普遍的炎症反应的结果。 4-NQO 模型将用于更广泛地了解嗜酸性粒细胞在肿瘤发生中的作用， 我们开发的复杂共培养技术提供了对嗜酸性粒细胞的机制见解 在肿瘤生物学中的激活和功能在该提案中，我们展示了嗜酸性粒细胞与食管共培养。 细胞增加凋亡，降低活力，并改变上皮细胞的细胞因子谱，增加 TNF-a 和 IL-2、IL-8 和 IL-10 的表达，以及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，这是显着的。 因为已知嗜酸性粒细胞过氧化物酶会催化卤化物和硫氰酸盐的氧化，从而导致释放 总而言之，这些数据表明嗜酸性粒细胞可能会降低活力。 ESCC 细胞通过释放 ROS 来表达，这是一种丰富的抗氧化酶。 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。 独立提出Gpx1调节嗜酸性粒细胞ROS的产生，以及ROS表达的减少 与食管上皮细胞共培养后，嗜酸性粒细胞检测到的 Gpx1 表明它被嗜酸性粒细胞修饰。 根据这些数据和嗜酸性粒细胞特异性向发育异常区域的募集。 和食管鳞癌中的癌症，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，并且 因此对 ESCC 具有保护作用。该项目的具体目标是： 1. 确定嗜酸性粒细胞的作用。 我们将利用 ESCC 的 4-NQO 模型来比较食管肿瘤的发生。 Ccl11-/- 与 4-NQO 治疗后的 WT 小鼠相比，我们将确定嗜酸性粒细胞耗竭的影响。 2. 确定不同时间点用 4-NQO 治疗的 WT 小鼠的嗜酸性粒细胞增多症的效果。 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的影响。 更具体地说，我们将定义嗜酸性粒细胞与小鼠类器官共培养的发育不良上皮。 通过将 Gpx1-/- 嗜酸性粒细胞与野生型共培养，Gpx1 对嗜酸性粒细胞功能和激活的作用 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措施。