P53 and KRAS Targeted Pigs: A Platform for Models of Human Cancer

P53 和 KRAS 靶向猪：人类癌症模型的平台

基本信息

批准号：
8314714
负责人：
Christopher Rogers
金额：
$ 16.71万
依托单位：
EXEMPLAR GENETICS, LLC
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：
2012-09-01 至 2013-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8314714
关键词：
Affect Alleles American Anatomy Animal Model Apoptosis Biological Models Breeding Cell Count Cell Cycle Arrest Cell Proliferation Cell physiology Cells Clinic Colon Couples Development Diagnosis Diagnostic Disease Engineering Environmental Risk Factor Family suidae Fetus Fibroblasts Frequencies Gene Targeting Generations Genes Genetic Genome Goals Harvest Human Industry KRAS2 gene Lung Adenocarcinoma Malignant Neoplasms Malignant neoplasm of lung Mission Modeling Monomeric GTP-Binding Proteins Mutate Mutation Neoplasm Metastasis Nuclear Oncogenes Pancreas Pancreatic Ductal Adenocarcinoma Pathogenesis Phase Physiology Production Receptor Activation Recombinant adeno-associated virus (rAAV)Research Personnel Resources Southern Blotting TP53 gene Testing Therapeutic Therapeutic Intervention Tissues Translating United States Work abstracting cancer type cell type design fetal homologous recombination instrument killings mouse model novel diagnostics somatic cell nuclear transfer tool transcription factor tumor tumor growth tumorigenesis tumorigenic vector

项目摘要

Project Summary/Abstract Cancer is the second deadliest disease in the United States, killing more than 500,000 Americans annually. This year, another 1.5 million will be diagnosed with one of nearly 200 different cancer types. Despite an ever---growing understanding of the environmental risk factors, genetic contributions, and tumorigenic mechanisms, the diagnoses and treatments for this disease remain inadequate. Much of what is known about cancer has come from studies of cells in culture and small animal models. While these model systems have been extremely informative, they also possess limitations and present challenges to translating promising therapies to the clinic. The lack of a large animal model that accurately replicates human cancer has been a major barrier to the development of effective diagnostic tools, interventions, and therapies for this deadly disease. Pigs share many similarities with humans in anatomy, physiology, genetics, and importantly, size. While naturally occurring tumors are rarely seen due to standard pork production practices, spontaneous and induced cancers have been studied in pigs and are highly representative of what is seen in humans. However, the extended timeframe required for inducing cancer in pigs and the accompanying phenotypic variability limit their usefulness. Our objective is to create a genetically engineered pig in which tumorigenesis can be conditionally induced in any tissue. We intend to accomplish this by mutating two of the most commonly affected genes in human cancer, KRAS and TP53 (encoding p53). KRAS is in oncogene encoding a small GTPase that couples receptor activation and downstream effectors to control cell proliferation, differentiation and survival. Activating mutations in KRAS are common in many human tumors. Known as the "guardian of the genome", p53 is a transcription factor that regulates critical cell functions including cell---cycle arrest and apoptosis. The loss of proper p53 function predisposes cells to unregulated growth, tumor formation, and metastasis. Mouse models expressing conditional mutations in KRAS and TP53 have yielded excellent models representative of human cancers of the lung, pancreas, colon, and other tissues. We hypothesize that mutations in porcine KRAS and TP53 will produce similar results in pigs. Therefore, the ultimate goal of this project is to develop and commercialize KRAS/TP53--mutated pigs to serve as a platform for models of human cancer. We intend to accomplish this by combining gene targeting and somatic cell nuclear transfer. This proposal specifically outlines the development of porcine fibroblasts with mutated KRAS and TP53 alleles. A gene targeting vector will be developed and used to disrupt the endogenous porcine KRAS via homologous recombination in both TP53---targeted and wild---type cells. Subsequent work will use these cells as nuclear donors for somatic cell nuclear transfer to produce KRAS/TP53---mutated pigs. These models will provide academic and industry researchers with an opportunity to better understand cancer and its pathogenesis and to develop and test new diagnostic, therapeutic, and preventative strategies.

项目摘要/摘要癌症是美国第二个致命的疾病，每年造成50万以上的美国人。今年，另外150万人将被诊断出近200种不同的癌症类型之一。尽管对环境危险因素，遗传贡献和肿瘤性机制的了解越来越不断提高，但该疾病的诊断和治疗仍然不足。关于癌症的许多知识来自对培养和小动物模型中细胞的研究。尽管这些模型系统非常有用，但它们也具有局限性，并提出了将有希望的疗法转化为诊所的挑战。缺乏准确复制人类癌症的大型动物模型是开发这种致命疾病有效诊断工具，干预措施和疗法的主要障碍。猪在解剖学，生理学，遗传学以及重要的是大小中与人类具有许多相似之处。尽管由于标准的猪肉生产实践而很少看到天然发生的肿瘤，但已经在猪中研究了自发和诱发的癌症，并且高度代表了人类所见。但是，诱导猪癌所需的延长时间范围以及随附的表型变异性限制了其有用性。我们的目标是创建一个基因工程的猪，其中可以在任何组织中有条件地诱导肿瘤发生。我们打算通过突变人类癌症，KRAS和TP53（编码p53）中的两个最常见的基因来实现这一目标。 KRAS正在编码一个小的GTPase的癌基因中，该GTPase将受体激活和下游效应子与控制细胞增殖，分化和存活。在许多人类肿瘤中，KRAS激活突变是常见的。 p53被称为“基因组的监护人”，是调节关键细胞功能在内的转录因子---循环停滞和凋亡。适当的p53功能的丧失使细胞易于生长，肿瘤形成和转移。在KRAS和TP53中表达条件突变的小鼠模型产生了代表肺，胰腺，结肠和其他组织的人类癌症的出色模型。我们假设猪KRAS和TP53中的突变会在猪中产生相似的结果。因此，该项目的最终目标是开发和商业化KRAS/TP53的猪，以作为人类癌症模型的平台。我们打算通过结合基因靶向和体细胞核转移来实现这一目标。该提案专门概述了具有突变的KRAS和TP53等位基因的猪成纤维细胞的发展。将开发靶向载体的基因靶向载体，并用于通过TP53中的同源重组来破坏内源性猪KRAS-靶向和野生 - 型细胞。随后的工作将将这些细胞用作核供体进行体细胞核转移，以产生KRAS/TP53 ---突变的猪。这些模型将为学术和行业研究人员提供更好地了解癌症及其发病机理的机会，并开发和测试新的诊断，治疗和预防策略。