Pathological Evaluation of a Cystic Fibrosis Ferret Model

囊性纤维化雪貂模型的病理学评估

基本信息

批准号：
7814270
负责人：
JOHN F ENGELHARDT
金额：
$ 49.95万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7814270
关键词：
Address Age Amino Acid Sequence Animal Model Animals Antibody Formation Area Bioinformatics Biology Birth Breeding Caring Caucasians Caucasoid Race Cellular biology Cessation of life Chloride Channels Chloride Ion Chlorides Chronic Cloning Communities Complementary DNA Coupled Cyclic AMP Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Databases Daughter Defect Development Disease Disease Progression Disease model Educational process of instructing Evaluation Exocrine pancreatic insufficiency Expressed Sequence Tags Family suidae Farming environment Fathers Female Ferrets Fetus Fibroblasts Future Gene Expression Profile Gene Targeting Genes Genetic Genetic Variation Genetically Engineered Mouse Genotype Goals Hereditary Disease Human Ileus Inbreeding Infant Infection Inflammation Information Technology Intervention Intestinal Obstruction Intestines Ions Knock-out Lesion Life Light Liquid substance Lung Marshal Martes zibellina Meconium Mediating Medical Microarray Analysis Modeling Molecular Mus National Institute of Diabetes and Digestive and Kidney Diseases Newborn Infant Nuclear Open Reading Frames Organ Orthologous Gene Other Genetics Outcome Pancreas Pathogenesis Pathology Penetrance Peptide Sequence Determination Phenotype Process Property Proteins Proteome Protocols documentation Pulmonary Cystic Fibrosis Reading Reagent Research Resources Sequence Alignment Somatic Cell Specimen Study models Techniques Technology Testing Time Tissues Titanium Transgenic Animals Transgenic Model Transgenic Organisms Translational Research Viral Work airway epithelium animal cloning base clinical care cohort cost cystic fibrosis mouse cystic fibrosis patients disease phenotype early experience fatty acid-binding proteins genetic strain human disease improved in vivo influenzavirus interdisciplinary approach intestinal fatty acid binding protein knockout animal novel promoter public health relevance reproductive somatic cell nuclear transfer

项目摘要

DESCRIPTION (provided by applicant): This application addresses two broad Challenge Areas and three associated specific Challenge Topics including: i) the broad Challenge Area (06) Enabling Technologies, specific Challenge Topic 06-HL- 105: Develop transgenic animal models that are informative for understanding chronic inflammation in humans, and specific Challenge Topic 06-HL-109: Generate reagents for studying lung cell biology and disease progression; and ii) the broad Challenge Area (15) Translational Science, and specific Challenge Topic: 15-DK-102 Develop improved animal models of NIDDK diseases. Cystic Fibrosis (CF) is the most common autosomal recessive disease in Caucasians and is caused by defects in the CFTR chloride channel. Many questions regarding the pathogenesis of this disease remain, in large part due to the lack of a CF animal model that mimics progression of human CF lung disease and exocrine pancreatic insufficiency (CFTR deficient mice lack both these properties). The ferret is an attractive species for modeling CF because: its lung biology is similar to that of humans, its reproductive time is rapid, and it can now be genetically manipulated. The ferret is also the preferred model organism for studying several types of human viral lung infection-most notably infection by influenza virus, which is thought to accelerate the progression of CF lung disease. We have generated a CFTR-deficient model in the ferret by performing somatic cell nuclear transfer with CFTR gene-targeted ferret fibroblasts. Preliminary analysis of CFTR-deficient newborn ferrets demonstrates that they are afflicted by meconium ileus, defects in cAMP-mediated chloride transport in the airway epithelium, and defects in pancreatic excocrine activity-abnormalities also seen in newborn patients with CF. This proposal seeks to characterize pathologies in a newly generated CFTR knockout ferret model, with a focus on enabling its use as a model for studying pathophysiologic mechanisms and therapies in the CF lung. Working closely with Marshall Farms (the major industrial breeder of ferrets in the U.S.), we have established a collaborative relationship that will allow us to achieve these goals in a cost-effective and timely manner. The outcome of this project-initial characterization of CF ferrets, establishment of a new genetic background for easier propagation without intestinal obstruction, and establishment of a corporate partnership for breeding and disseminating this model to the research community-will greatly impact research on the multi-organ aspects of CF disease. Equally important to the characterization of this model is the development of a ferret molecular toolbox. We propose to tackle this issue while also addressing biologic questions about early transcriptional changes within the CF pancreas and lung. 454 GS FLX Titanium deep sequencing technology will be used for transcriptome analyses, giving rise to 3.6-6 Mb of new EST sequence information on the uncharted ferret transcriptome. This information will enable microarray technologies in this species, and the production of antibodies to ferret proteins, thereby greatly facilitating the use of the ferret as a disease model for research. PUBLIC HEALTH RELEVANCE: Cystic Fibrosis (CF) is the most common life-threatening autosomal recessive condition among Caucasians with over $450 million dollars spent on clinical care of CF patients annually in the U.S. alone. Despite the fact that the gene defect responsible for CF was discovered over 20 years ago, a cure for CF has yet to emerge. In large part, this is due to the lack of good animal models that reproduce the human CF disease phenotype on which to both understand the disease process and test therapies. This proposal is focused on the initial characterization and further development of a new animal model of CF in the ferret. This model shows classic signs of human CF disease and may be a valuable resource for the CF research community following completion of the proposed project.

描述（由申请人提供）：本申请涉及两个广泛的挑战领域和三个相关的具体挑战主题，包括：i) 广泛的挑战领域 (06) 实现技术，具体挑战主题 06-HL-105：开发信息丰富的转基因动物模型用于了解人类慢性炎症，以及具体的挑战主题 06-HL-109：生成用于研究肺细胞生物学和疾病进展的试剂； ii) 广泛的挑战领域 (15) 转化科学，以及具体的挑战主题：15-DK-102 开发改进的 NIDDK 疾病动物模型。囊性纤维化 (CF) 是白种人中最常见的常染色体隐性遗传疾病，由 CFTR 氯离子通道缺陷引起。关于这种疾病发病机制的许多问题仍然存在，很大程度上是由于缺乏模拟人类 CF 肺病和外分泌胰腺功能不全进展的 CF 动物模型（CFTR 缺陷小鼠缺乏这两种特性）。雪貂是用于 CF 建模的一个有吸引力的物种，因为：它的肺部生物学与人类相似，它的繁殖时间很快，而且现在可以进行基因操纵。雪貂也是研究几种人类病毒性肺部感染的首选模型生物，尤其是流感病毒感染，人们认为流感病毒会加速 CF 肺部疾病的进展。我们通过用 CFTR 基因靶向的雪貂成纤维细胞进行体细胞核移植，在雪貂中生成了 CFTR 缺陷模型。对 CFTR 缺陷的新生雪貂的初步分析表明，它们患有胎粪性肠梗阻、气道上皮中 cAMP 介导的氯化物转运缺陷以及胰腺外分泌活动缺陷，这些缺陷在 CF 新生儿患者中也可见到。该提案旨在描述新生成的 CFTR 敲除雪貂模型的病理特征，重点是使其能够用作研究 CF 肺部病理生理机制和治疗的模型。我们与马歇尔农场（美国主要的雪貂工业饲养场）密切合作，建立了合作关系，这将使我们能够以具有成本效益的方式及时实现这些目标。该项目的成果——CF 雪貂的初步表征、建立新的遗传背景以更容易繁殖而无肠梗阻，以及建立企业合作伙伴关系以培育并向研究界传播该模型——将极大地影响多方面的研究。 CF 疾病的器官方面。与该模型的表征同样重要的是雪貂分子工具箱的开发。我们建议解决这个问题，同时解决有关 CF 胰腺和肺内早期转录变化的生物学问题。 454 GS FLX Titanium 深度测序技术将用于转录组分析，在未知的雪貂转录组上产生 3.6-6 Mb 的新 EST 序列信息。这些信息将使微阵列技术应用于该物种，以及雪貂蛋白抗体的生产，从而极大地促进雪貂作为疾病模型的研究。公共健康相关性：囊性纤维化 (CF) 是白种人中最常见的危及生命的常染色体隐性遗传病，仅在美国，每年用于 CF 患者临床护理的费用就超过 4.5 亿美元。尽管导致 CF 的基因缺陷在 20 多年前就已被发现，但 CF 的治疗方法尚未出现。这在很大程度上是由于缺乏能够重现人类囊性纤维化疾病表型的良好动物模型，以了解疾病过程并测试治疗方法。该提案的重点是雪貂 CF 新动物模型的初步表征和进一步开发。该模型显示了人类 CF 疾病的典型症状，在拟议项目完成后可能成为 CF 研究界的宝贵资源。