Single Somatic Cell Epigenetic Models

 描述(申请人提供)：DNA和组蛋白的表观遗传修饰是生命必不可少的稳定加密系统。例如，DNA甲基化的重要性被其错误调控的后果所强调，其中包括组织稳态缺陷、慢性病和加速衰老；因此，表观遗传学广泛影响人类健康。存储在5-甲基胞嘧啶分布中的信息在细胞分裂后是灵活、稳定和可遗传的；然而，与遗传突变不同的是，表观遗传变化可能在药理学上被逆转。这项研究的目标是将产品商业化，这些产品将用于更好地了解疾病中表观遗传失误的演变，并发现新的表观治疗学。我们提出了一系列基于荧光色素的报告试剂盒，内容丰富，可检查人类疾病的表观遗传重编程(印迹)和DNA甲基化维持。这些产品将在发现癌症、帕金森氏病、炎症性肠道疾病的治疗策略方面得到应用，以及在人类和非人类灵长类动物中发展对新发现的表观基因组时钟(衰老)的理解。这将通过确定可用药的靶点，并剖析调节DNA甲基化老化和慢性疾病的途径蛋白，促进新的治疗进展。这些商业产品基于一个模型系统，该系统使用内源性GFP/RFP报告程序和归巢内切酶通过同源和非同源途径介导的损伤修复；这两种途径都会在修复的DNA片段上诱导甲基化修改，并产生新的表观等位基因。由于这是一种基于细胞的印记，因此将实现两个重要的好处。第一，高分辨率，单细胞表观遗传学将成为可能；第二，单个细胞可以被追踪 通过实时成像传递给后代。该公司将提供一系列记者(表达载体、特别工程细胞系和试剂盒)来检查甲基化修改中的变化和确定的疾病模型中的修复。这将允许在细胞范围内对新的表观疗法进行创新的筛选系统。总而言之，这项建议将DNA甲基化维持的基本信息转化为一个灵活的系统，可以用于监测、操纵和剖析体细胞和动物模型中的甲基化过程。这些产品将用于生物医学研究的不同领域(关于环境与表观遗传回路、神经学、炎症、代谢性疾病、表观遗传诱变剂、慢性多性状疾病、营养、衰老等的研究)。对于第二阶段SBIR，我描述了一种新的生物信息学算法，它解释了表型进化在临床和个性化医学中的应用

 DESCRIPTION (provided by applicant): Epigenetic modifications of DNA and histones are stable encryption systems essential for life. For example, the importance of DNA methylation is underscored by the consequences of its mis-regulation, which include defects in tissue homeostasis, chronic disease and accelerated aging; thus epigenetics broadly impacts human health. The information stored in the distribution of 5-methyl-cytosine is flexible, stable and heritable after cell division; however, unlike genetic mutation, epigenetic changes may be pharmacologically reversed. The goal of this research is to commercialize products that will be used to develop a better understanding of the evolution of epigenetic miscues in disease and to discover new epi-therapuetics. We propose a series of fluorochrome- based reporter kits with content rich protocols that examine epigenetic reprogramming (imprinting) and DNA methylation maintenance in human disease. Such products will see applications in discovery of treatment strategies in cancer, Parkinson's disease, Inflammatory Bowel Disease, as well as developing an understanding behind the newly discovered epigenomic clock (aging) in human and non-human primates. This will promote new therapeutic inroads by identifying druggable targets, and dissecting pathway proteins regulating DNA methylation aging and chronic disease. The commercial products are based on a model system that uses endogenous GFP/RFP reporters and homing endonuclease mediated damage-repair by homologous and non- homologous routes; both pathways induce methylation revision at repaired DNA segments and yield new epialleles. Because this is a cell based imprinting, two important benefits will be realized. First high resolution, single cell epigenetics will be possible; and second, a single cell can be tracked into future generations by live imaging. The company will offer a selection of reporters (expression vectors, ad hoc engineered cell lines and kits) to examine alterations in methylation revisions and repair in defined disease models. This will allow an innovative screening system for novel epi- therapeutics in a cellular context. In sum, this proposal translates the basic information on DNA methylation maintenance into a flexible system that can be used to monitor, manipulate and dissect the process in somatic cells and animal models. These products will be used in diverse areas of biomedical research (studies on environmental connectivity to epigenetic circuitry, neurologic, inflammatory, metabolic diseases, epigenetic mutagens, chronic multi-trait diseases, nutrition, aging, etc). For the Phase II SBIR a new bioinformatic algorithm i described that interprets epiallele evolution for application in clinical and personalized medicine