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：
8415397
负责人：
JOHN P. LYNCH
金额：
$ 37.56万
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 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8415397
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. PUBLIC HEALTH RELEVANCE: Chronic inflammatory conditions are an especially burdensome healthcare problem in the US. Not only must the pain and disability be managed, but patients and health-care providers must be vigilant for the long-term consequences of chronic inflammation, most prominently cancer. Two examples of this are acid reflux disease, which leads to metaplasia and cancer, and chronic inflammatory conditions of the intestine and colon like graft vs. host disease or ulcerative colitis, the latter which can also frequently progress to colon cancer. At present there is little known about the mechanisms giving rise to metaplasia and cancer in the setting of chronic inflammation. This proposal describes approaches to model and mechanistically explore the contributions of Cox-2, oxidative stress and DNA damage, inflammatory cells, and epithelial cells to the pathogenesis of metaplasia and cancer, and we anticipate our approaches will yield improved human cell culture models suitable for testing and 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，例如胃食管反流病（GERD），肠道移植与宿主疾病（GVHD），小肠移植后的急性抑制，以及诸如溃疡性Coliser Colisation Colisation Colisation Colisation Colisation（UC）。基于人类细胞和组织培养系统概括体内生长和分化模式的疾病模型将增强我们对疾病进展的理解，并改善预防和检测策略。这是该提议的重要目标。在这些疾病中，上皮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诱导DNA损伤和突变的内源性酶（环氧酶和NADPH氧化酶），并改变了DNA甲基化模式，这共同有助于化生和癌症的发展。我们已经开始使用器官型培养系统探索这一点，以在发病机理中对COX-2建模。当我们在正常人类食管角质形成细胞中异位表达COX-2时，我们会观察到肠粘膜填充囊肿的发展。我们建议将这种成功扩展在生理相关的方向上。我们的主要目标是检验以下假设：可修改器官型培养系统以模拟食管和肠上皮中的急性和慢性氧化应激，这些培养系统在生理上类似于GERD，BE，GVHD，GVHD，小肠子移植物和UC的体内事件中与体内事件相似。我们建议通过追求以下特定目的来测试这一点：1）使食管型器官培养系统改善GERD食管炎，并将干细胞的进展到化生和发育不良。 2）开发器官和3D多细胞培养系统，以建模GVHD的炎症微环境，小肠移植抑制和包括UC在内的IBD。 GERD/BE，IBD/NEOPLASIA和GVHD是重要的，相对常见的条件，会给美国医疗保健系统带来巨大负担。我们建议开发新型的多细胞体外人体组织工程模型，这些模型代表了这些条件的发病机理。这些模型将具有巨大的价值，使我们能够检验假设并迅速提高我们对这些疾病的理解，并且由于药物对COX-2的抑制作用已得到充分确立，因此会产生翻译影响。这项工作将大大提高我们学习，预防和治疗巴雷特食管，IBD和GVHD的能力。它还将促进新的治疗和预防策略的发展，以改善这些重要的临床状况的患者护理。公共卫生相关性：在美国，慢性炎症状况是一个特别繁重的医疗保健问题。不仅必须管理疼痛和残疾，而且患者和医疗保健提供者必须警惕慢性炎症的长期后果，最突出的癌症。这两个例子是胃酸反流疾病，导致化生和癌症，以及肠道和结肠的慢性炎症状况，例如移植物与宿主疾病或溃疡性结肠炎，后者也可以经常发展为结肠癌。目前，关于机制鲜为人知在慢性炎症的情况下引起了化生和癌症。该建议介绍了模型和机械探索COX-2，氧化应激和DNA损伤，炎性细胞以及上皮细胞对化生和癌症发病机理的贡献的方法，我们预计我们的方法将产生适合于这些重要的患者护理条件的治疗和预防策略的人类细胞培养模型。