Antioxidant regulation of intestinal homeostasis and disease

肠道稳态和疾病的抗氧化调节

• 批准号: 10334557
• 负责人: Sarah Palmer Short
• 金额: $ 14.53万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10334557
• 关键词: 3-Dimensional; Active Sites; Adenocarcinoma; Affect; Amino Acids; Animals; Antioxidants; Attenuated; Bone Marrow; Bone Marrow Transplantation; Carcinoma; Cell Culture Techniques; Cell physiology; Cells; Cellular biology; Colitis; Complement; Crohn' s disease; Cytoplasm; Data; Development; Disease; Doctor of Philosophy; Dysplasia; Ensure; Epidemiology; Epithelial; Epithelial Cells; FDA approved; Flow Cytometry; GPX2 gene; Gastroenterology; Gastrointestinal Diseases; Goals; Hematopoietic; Homeostasis; Hydrogen; Ileocolitis; Immune; Immune response; Immunological Models; Immunology; In Vitro; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Injury; Institution; Intestinal Diseases; Intestines; Lead; Malignant Neoplasms; Mediating; Medicine; Mentors; Metabolism; Micronutrients; Mitochondria; Modeling; Mucous Membrane; Mus; Natural regeneration; Organoids; Oxidation-Reduction; Oxidative Stress; Patients; Proteins; Reduced Glutathione; Regulation; Research; Research Activity; Research Personnel; Role; Selenium; Selenocysteine; Severities; Severity of illness; Single Nucleotide Polymorphism; Sodium Dextran Sulfate; Stimulus; T-Lymphocyte; Testing; Therapeutic; TimeLine; Training; Ulcerative Colitis; Work; base; career; career development; chemokine; colitis associated cancer; dextran sulfate sodium induced colitis; disorder risk; epithelial injury; experience; experimental study; glutathione peroxidase; improved; in vivo; intestinal epithelium; intestinal homeostasis; intestinal injury; macrophage; murine colitis; programs; protective effect; recruit; response; response to injury; selenium deficiency; selenoenzyme; selenoprotein; stem cell function; stem cells; stemness; therapeutic target; therapeutically effective; tissue/cell culture; wound healing

Project Summary This proposal details a 5-year training plan to aid the continued develop of Dr. Sarah Short, Ph.D. into an independent GI researcher. This research plan will focus on elucidating the role of glutathione peroxidase 1 (GPx1), a ubiquitously expressed selenoenzyme and potent antioxidant, in inflammatory bowel disease (IBD) and colitis-associated dysplasia (CAD). Compelling preliminary data using Gpx1-/- mice indicates that unlike many antioxidants whose loss exacerbates murine colitis, loss of GPx1 confers striking protection from dextran- sodium sulfate (DSS)-induced colitis. GPx1 deficiency also increases survival and stemness in 3D organoids and alters metabolism in tissue culture cells, which may additionally promote regeneration and wound healing. GPx1 expression also modifies immune cell function, as Gpx1-/- bone marrow-derived macrophages have heightened response to “M2” stimuli and decreased migratory ability. Together, these results suggest that GPx1 augments inflammatory injury through alterations in both epithelial and immune cell function. Based on these finding, the hypothesis of this proposal is that GPx1 is detrimental in inflammatory bowel disease by altering stem cell function, redox homeostasis, and immune responses. Further, inhibiting GPx1 activity may be an effective therapeutic strategy. This hypothesis will be tested in two specific aims to determine how GPx1 contributes to intestinal epithelial cell homeostasis, oxidative stress, colitis, and colitis-associated dysplasia. The first aim will investigate epithelial function, capitalizing on Dr. Short’s over 10 years of experience in epithelial cell biology and barrier function. The second aim will complement epithelial-based studies by determining how GPx1 loss alters immune cell recruitment, differentiation, and function, and identify how these changes modify intestinal injury responses. In addition to being the logical “next step” experiments in defining GPx1 function, these experiments provide the perfect framework to further Dr. Short’s development in aspects of mucosal immunology which contribute to intestinal diseases, and will include new training in flow cytometry, chemokine analysis, bone marrow transplantation, and the T-cell transfer colitis model. Dr. Short’s career development will be further enhanced by regular discussions with primary mentor, Dr. Christopher Williams, and her mentoring committee consisting of Drs. Keith Wilson, Jeremy Goettel, and Sean Davies. All studies and training will take place at Vanderbilt, and the institution, Department of Medicine, and Gastroenterology Division are highly supportive of Dr. Short’s academic career and fully support her application. Dr. Short’s ultimate goal is to become an independent academic researcher focusing on mechanisms which regulate development and severity of IBD and colitis-associated cancer that can lead to improved therapeutic options for these patients. Interestingly, both specific aims proposed in this application will evaluate GPx1 as a therapeutic target using tiopronin, which is FDA-approved and well-tolerated. Together, these training experiences will ensure Dr. Short is poised to direct a well-rounded independent research program in IBD.

项目概要 该提案详细介绍了一项为期 5 年的培训计划，以帮助 Sarah Short 博士的持续发展。进入一个 独立地理标志研究员。该研究计划将重点阐明谷胱甘肽过氧化物酶 1 的作用 (GPx1) 是一种在炎症性肠病 (IBD) 中普遍表达的硒酶和有效抗氧化剂 和结肠炎相关发育不良（CAD）。使用 Gpx1-/- 小鼠获得的令人信服的初步数据表明，与 许多抗氧化剂的丢失会加剧小鼠结肠炎，GPx1 的丢失可提供对葡聚糖的显着保护 硫酸钠（DSS）诱发的结肠炎。 GPx1 缺乏也会增加 3D 类器官的存活率和干性 并改变组织培养细胞的新陈代谢，这可能另外促进再生和伤口愈合。 GPx1 表达还会改变免疫细胞功能，因为 Gpx1-/- 骨髓来源的巨噬细胞具有 对“M2”刺激的反应增强，迁移能力下降。总之，这些结果表明 GPx1 通过改变上皮细胞和免疫细胞功能来增强炎症损伤。 基于这些发现，该提案的假设是 GPx1 对炎症性肠有害 通过改变干细胞功能、氧化还原稳态和免疫反应来预防疾病。此外，抑制GPx1 活动可能是一种有效的治疗策略。该假设将在两个具体目标中进行测试以确定 GPx1 如何促进肠上皮细胞稳态、氧化应激、结肠炎和结肠炎相关疾病 发育不良。第一个目标是利用 Short 博士 10 多年的经验来研究上皮功能 上皮细胞生物学和屏障功能。第二个目标将补充基于上皮的研究 确定 GPx1 丢失如何改变免疫细胞的招募、分化和功能，并确定这些如何改变 变化会改变肠道损伤反应。除了定义逻辑上的“下一步”实验之外 GPx1功能，这些实验为Short博士进一步发展提供了完美的框架 粘膜免疫学有助于肠道疾病，并将包括流式细胞术的新培训， 趋化因子分析、骨髓移植和 T 细胞转移结肠炎模型。肖特博士的职业生涯 通过与主要导师克里斯托弗·威廉姆斯博士的定期讨论，将进一步促进发展， 她的指导委员会由博士组成。基思·威尔逊、杰里米·戈特尔和肖恩·戴维斯。所有研究和 培训将在范德比尔特大学、该机构、医学系和胃肠病学部门进行 高度支持肖特博士的学术生涯并全力支持她的申请。 肖特博士的最终目标是成为一名独立的学术研究员，专注于研究机制 调节 IBD 和结肠炎相关癌症的发展和严重程度，从而改善治疗 这些患者的选择。有趣的是，本申请中提出的两个具体目标都将 GPx1 评估为 使用硫普罗宁作为治疗靶点，硫普罗宁已获得 FDA 批准且耐受性良好。这些训练共同 丰富的经验将确保 Short 博士能够指导 IBD 领域全面的独立研究项目。