Modeling oxidative stress and DNA damage using GI organotypic culture systems

使用胃肠道器官培养系统模拟氧化应激和 DNA 损伤

基本信息

批准号：
8697174
负责人：
JOHN P. LYNCH
金额：
$ 8万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-24 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8697174
关键词：
Acids Acute Acute Graft Versus Host Disease Adoption Adoptive Transfer Allogeneic Bone Marrow Transplantation American Anabolism BMP4 Barrett Esophagus Bone Marrow Transplantation Cell Culture System Cell Culture Techniques Cell Lineage Cell Proliferation Cells Chronic Clinical Colon Colon Carcinoma Cox Models Cyst DNA Damage DNA Methylation Detection Development Differentiation and Growth Dinoprostone Disease Disease Progression Disease model Dysplasia Effector Cell Eicosanoids Enterocytes Environment Enzymes Epigenetic Process Epithelial Epithelial Cells Epithelium Esophageal Esophagitis Esophagus Event Experimental Models Fibroblasts Fostering Gastroesophageal reflux disease Gastrointestinal tract structure Gene Expression Gene Expression Profile Gene Mutation Graft Rejection Growth Health Personnel Healthcare Healthcare Systems Human Immune In Vitro Inflammation Inflammatory Inflammatory Response Intestines Malignant Neoplasms Mediating Metaplasia Metaplastic Modeling Morphology Mucins Mus Mutation NADPH Oxidase Neoplasms Oncogenes Oxidative Stress Pain Pathogenesis Patient Care Patients Pattern Phenotype Prevention Prevention strategy Process Production Prostaglandin-Endoperoxide Synthase Prostaglandins Reflux Small Intestines Stem cells System T-Lymphocyte Testing Tissue Engineering Tissue Model Transplantation Ulcerative Colitis Work base cytokine disability graft vs host disease human disease human tissue improved in vivo innovation intestinal epithelium keratinocyte novel novel therapeutics oxidative DNA damage prevent response success tissue culture tissue/cell culture

项目摘要

DESCRIPTION (provided by applicant): Acute and chronic inflammatory environments of the gastrointestinal tract like gastroesophageal reflux disease (GERD), intestinal graft vs. host disease (GVHD), acute rejection after small intestine transplantation, and IBD conditions like Ulcerative colitis (UC), are fairly common human diseases. Disease models based on human cell and tissue culture systems that recapitulate in vivo growth and differentiation patterns would enhance our understanding of disease progression and improve prevention and detection strategies. This is an important objective of this proposal. In these disorders, epithelial cel oxidative stress is a key pathogenic factor for disease progression. This oxidative stress is partly from endogenous enzymes (Cyclooxygenases and NADPH oxidases) that induce DNA damage and mutations, and alter DNA methylation patterns, which together contributes to the development of metaplasia and cancer. We have begun to explore this using organotypic culture systems to model Cox-2 in BE pathogenesis. When we ectopically express Cox-2 in normal human esophageal keratinocytes, we observe the development of intestinal mucin-filled cysts. We propose to extend this success in physiologically relevant directions. Our main objective will be to test the hypothesis that the organotypic culture systems can be modified to model acute and chronic oxidative stress in esophageal and intestinal epithelium that physiologically resembles in vivo events in GERD, BE, GVHD, small bowel transplant rejection, and UC. We propose to test this by pursuing the following Specific Aims: 1) Adapt the esophageal organotypic culture system to better model GERD esophagitis and progression of stem cells to metaplasia and dysplasia. 2) Develop organotypic and 3D multi-cellular culture systems to model inflammatory microenvironments of GVHD, small bowel transplant rejection, and IBD including UC. GERD/BE, IBD/neoplasia, and GVHD are important, relatively common conditions that place a significant burden on the US healthcare system. We propose to develop novel multi-cellular in vitro human tissue engineered models that are representative of the pathogenesis for these conditions. These models will be of enormous value, allowing us to test hypotheses and advance our understanding of these disorders rapidly, and would have a translational impact since pharmacologic inhibition of Cox-2 is well established. This work will greatly improve our ability to study, prevent, and treat Barrett's esophagus, IBD, and GVHD. It will also foster the development of novel therapeutic and preventive strategies that will improve patient care for these important clinical conditions.

描述（由申请人提供）：胃肠道的急性和慢性炎症环境，如胃食管反流病（GERD）、肠移植物抗宿主病（GVHD）、小肠移植后的急性排斥反应以及IBD病症，如溃疡性结肠炎（UC），是相当常见的人类疾病。基于人类细胞和组织培养系统的疾病模型可以概括体内生长和分化模式增强我们对疾病进展的了解并改进预防和检测策略。这是本提案的一个重要目标。在这些疾病中，上皮细胞氧化应激是疾病进展的关键致病因素。这种氧化应激是部分来自内源性酶（环加氧酶和 NADPH 氧化酶），它们会诱导 DNA 损伤和突变，并改变 DNA 甲基化模式，共同促进化生和癌症的发展。我们已经开始使用器官型培养系统来探索 BE 发病机制中的 Cox-2 模型。当我们在正常人食管角质形成细胞中异位表达 Cox-2 时，我们观察到肠道粘蛋白填充囊肿的形成。我们建议将这一成功扩展到生理相关方向。我们的主要目标是测试这样的假设：可以修改器官型培养系统来模拟食管和肠上皮的急性和慢性氧化应激，其生理学上类似于 GERD、BE、GVHD、小肠移植排斥和 UC 的体内事件。我们建议通过追求以下具体目标来测试这一点：1）调整食管器官型培养系统以更好地模拟 GERD 食管炎以及干细胞向化生和发育不良的进展。 2) 开发器官型和 3D 多细胞培养系统，以模拟 GVHD、小肠移植排斥和 IBD（包括 UC）的炎症微环境。 GERD/BE、IBD/肿瘤和 GVHD 是重要且相对常见的疾病，给美国医疗保健系统带来了沉重负担。我们建议开发新型多细胞体外人体组织工程模型，以代表这些病症的发病机制。这些模型将具有巨大的价值，使我们能够检验假设并迅速增进我们对这些疾病的理解，并且由于 Cox-2 的药理学抑制已得到充分证实，因此将产生转化影响。这项工作将极大提高我们研究、预防和治疗巴雷特食管、IBD 和 GVHD 的能力。它还将促进新型治疗和预防策略的发展，从而改善对这些重要临床病症的患者护理。