Single Somatic Cell Epigenetic Models

 描述（由申请人提供）：DNA和组蛋白的表观遗传修饰是生命所必需的稳定加密系统。例如，DNA甲基化的重要性是由其错误调节的后果所强调的，其中包括组织稳态缺陷，慢性疾病和加速衰老;因此表观遗传学广泛影响人类健康。细胞分裂后，5-甲基胞嘧啶分布中储存的信息是灵活的、稳定的和可遗传的;然而，与遗传突变不同，表观遗传变化可能是可逆的。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错误的演变，并发现新的表观治疗方法。我们提出了一系列基于荧光染料的报告试剂盒，内容丰富的协议，检查人类疾病中的表观遗传重编程（印记）和DNA甲基化维持。这些产品将应用于发现癌症，帕金森病，炎症性肠病的治疗策略，以及在人类和非人类灵长类动物中新发现的表观基因组时钟（衰老）背后的理解。这将通过确定可药物化的靶点，并解剖调节DNA甲基化衰老和慢性疾病的途径蛋白质来促进新的治疗进展。商业产品基于使用内源GFP/RFP报告基因和归巢核酸内切酶介导的通过同源和非同源途径的损伤修复的模型系统;两种途径均诱导修复的DNA区段处的甲基化修正并产生新的表观等位基因。因为这是基于细胞的印记，所以将实现两个重要的益处。第一，高分辨率，单细胞表观遗传学将成为可能;第二，单细胞可以被跟踪 传给后代该公司将提供一系列报告者（表达载体，特设工程细胞系和试剂盒），以检查定义的疾病模型中甲基化修订和修复的改变。这将允许在细胞环境中用于新型表观治疗剂的创新筛选系统。总之，该提案将DNA甲基化维持的基本信息转化为一个灵活的系统，可用于监测，操纵和解剖体细胞和动物模型中的过程。这些产品将用于生物医学研究的各个领域（研究环境与表观遗传回路、神经系统、炎症、代谢疾病、表观遗传诱变剂、慢性多性状疾病、营养、衰老等的联系）。对于第二阶段SBIR，我描述了一种新的生物信息学算法，该算法解释了表观等位基因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