Antioxidant regulation of intestinal homeostasis and disease

肠道稳态和疾病的抗氧化调节

• 批准号: 10553707
• 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 2023-04-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10553707
• 关键词: 3-Dimensional; Active Sites; Adenocarcinoma; Affect; Amino Acids; Animal Model; Antioxidants; Attenuated; Bone Marrow; Bone Marrow Transplantation; Carcinoma; Cell Culture Techniques; Cell physiology; Cells; Cellular biology; Colitis; Complement; Crohn' s disease; Cytoplasm; Data; Development; Doctor of Philosophy; Dysplasia; Ensure; Epidemiology; Epithelial Cells; Epithelium; FDA approved; Flow Cytometry; GPX2 gene; Gastroenterology; Gastrointestinal Diseases; Goals; Hematopoietic; Homeostasis; Hydrogen Peroxide; Ileocolitis; Immune; Immune response; Immunological Models; Immunology; In Vitro; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Injury; Institution; Intestinal Diseases; Intestines; Macrophage; 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; Training; Ulcerative Colitis; Work; career; career development; chemokine; cohort; 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; murine colitis; programs; protective effect; recruit; response; response to injury; selenium deficiency; selenoenzyme; selenoprotein; stem cell function; stem cells; stemness; therapeutic target; therapeutically effective; timeline; 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.

项目摘要 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 独立的GI研究员。该研究计划将着重阐明谷胱甘肽过氧化物酶1 （GPX1），一种普遍表达的硒酶和潜在的抗氧化剂，在炎症性肠病（IBD）中 和结肠炎相关的发育不良（CAD）。使用gpx1 - / - 小鼠引人入胜的初步数据表明，与 许多抗氧化剂的损失加剧了鼠结肠炎，GPX1的丧失给右旋 硫酸钠（DSS）诱导的结肠炎。 GPX1缺乏症也会增加3D器官中的存活率和干性 并改变组织培养细胞中的代谢，这可能会促进再生和伤口愈合。 GPX1表达也会修饰免疫电池功能，因为GPX1 - / - 骨髓来源的巨噬细胞具有 对“ M2”刺激的反应增强并提高了迁移能力。这些结果在一起表明GPX1 通过改变上皮和免疫功能来增强炎症损伤。 基于这些发现，该提议的假设是GPX1在炎症性肠中有害 通过改变干细胞功能，氧化还原稳态和免疫调查的疾病。此外，抑制GPX1 活动可能是一种有效的治疗策略。该假设将以两个特定的目的进行检验，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 发育不良。第一个目标将调查上皮功能，利用Short博士的10多年经验 在上皮细胞生物学和屏障功能中。第二个目标将通过 确定GPX1损失如何改变免疫细胞的募集，分化和功能，并确定这些方式如何 变化改变了肠道损伤反应。除了是定义的逻辑“下一步”实验 gpx1功能，这些实验为进一步的short博士的发展提供了完美的框架 有助于肠道疾病的粘膜免疫学，包括流式细胞仪的新训练， 趋化因子分析，骨髓移植和T细胞转移结肠炎模型。 Short博士的职业 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增强发展。 她的心理委员会由Drs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（Vanderbilt），医学院和胃肠病学部门进行 高度支持Short博士的学术生涯，并充分支持她的应用。 Short博士的最终目标是成为一名专注于机制的独立学术研究员 调节IBD和与结肠炎相关癌症的发育和严重程度，这可以改善治疗 这些患者的选择。有趣的是，本应用程序中提出的两个具体目标都将评估GPX1 使用Tiorponin的治疗靶标，该靶标是FDA批准且耐受良好的靶标。在一起，这些训练 经验将确保Short博士被中毒，以指导IBD的全面独立研究计划。