Epigenomic Regulation in Pancreatic Cell Growth

这项提案要求延长我们长期以来用于调查转录的主要资金来源， 染色质和表观遗传学在胰腺疾病的病理生理机制。我们的研究将 直接扩大我们对常见疾病的认识，如慢性胰腺炎和胰管炎， 胰腺癌（PDAC）是胰腺外分泌的疼痛性和不可治愈的疾病， 预防方案和治疗方法仍有待充分制定。我们的总体目标是 研究表观基因组调控因子如何作为与人类相关的常见突变的核效应子发挥作用 胰腺疾病虽然我们相信我们的概念和方法可以应用于任何 “突变-表观遗传-疾病三联体”，我们将重点了解组蛋白为基础的途径作为效应器， KRAS在已建立的胰腺疾病基因工程小鼠和人类患者源性模型中的应用 值得注意的是，虽然与遗传改变类似，表观遗传变化是可遗传的，但它们可以通过以下方式逆转： 药物治疗我们的初步数据报告发现组蛋白H3的关键作用 赖氨酸9（H3K9）甲基化途径及其相关甲基转移酶EHMT2作为表观遗传调节因子 致癌的KRAS EHMT2及其副产物EHMT1是主要的组蛋白赖氨酸甲基转移酶 负责催化组蛋白H3K9二甲基化。然而，没有关于这方面的信息。 EHMT1和EHMT2的功能，单独或作为复合物，与胰腺生理学或 胰腺疾病我们将检验一个综合的、机械的和翻译的中心假设，即 EHMT1/EHMT2复合物在ADM和PanIN形成过程中作为KRAS的表观遗传效应物发挥作用， 以及胰腺炎的进展。我们的具体目标是：1。确定机制， EHMT 12复合物的失活单独拮抗KRASG 12D介导的ADM和PanIN形成， 胰腺炎; 2.评估胰腺细胞中KRAS信号调节的生化机制 EHMT 1/2复合物的酶活性;和3.为了发现表观遗传机制， EHMT1/2复合物介导KRAS的作用。我们将使用大量的细胞，分子和 整个生物体实验，在高度协作的环境中使用最先进的技术执行。 通过专注于更好地理解表观基因组途径，作为共同的下游效应器， 突变的胰腺，我们的设计旨在最大限度地提高产量的快速翻译机制， 知识这项提案的发现将产生宝贵的积极影响，因为 来自表观基因组学领域的概念、技术和药物有望为以下方面提供新的机会： 胰腺疾病患者的管理，因此具有重要的生物医学意义。

This proposal requests the renewal of our major source of long-standing funding for investigating transcription, chromatin, and epigenetics in pathophysiological mechanisms underlying pancreatic diseases. Our studies will directly extend our knowledge on common diseases, such as chronic pancreatitis and pancreatic ductal adenocarcinoma (PDAC), both painful and incurable disorders of the exocrine pancreas for which effective prevention protocols and treatments remain to be fully developed. Our OVERALL OBJECTIVE is to investigate how epigenomic regulators work as nuclear effectors of common mutations associated with human pancreatic diseases. Although we believe that our concepts and methodologies can be applied to any “mutation-epigenetic-disease triad”, we will focus on understanding histone-based pathways as effectors of KRAS in established genetically-engineered mouse and human patient-derived models of pancreatic diseases. Notably, while similar to genetic alterations epigenetic changes are inheritable, they can be reversible by pharmacological treatment. Our PRELIMINARY DATA report the discovery of a key role for the Histone H3 Lysine 9 (H3K9) methylation pathway and its associated methyltransferase, EHMT2, as an epigenetic regulator of oncogenic KRAS. EHMT2 together with its paralog EHMT1 are the main histone lysine methyltransferases responsible for catalyzing histone H3K9 dimethylation. However, no information is known regarding the function of EHMT1 and EHMT2, either separately or as a complex, in relationship to pancreas physiology or pancreatic diseases. We will test a combined, mechanistic and translational CENTRAL HYPOTHESIS, namely that the EHMT1/EHMT2 complex works as an epigenetic effector of KRAS during ADM and PanIN formation as well as their progression by pancreatitis. Our SPECIFIC AIMS are: 1. To determine mechanisms by which inactivation of the EHMT1/2 complex antagonizes KRASG12D-mediated ADM and PanIN formation alone and in pancreatitis; 2. To evaluate biochemical mechanisms by which KRAS signaling in pancreatic cells regulates the enzymatic activity of the EHMT1/2 complex; and 3. To discover epigenetic mechanisms by which the EHMT1/2 complex mediates the effects of KRAS. We will use an extensive battery of cellular, molecular and whole organism experiments, executed in a highly collaborative environment with state-of-the-art techniques. By focusing on better understanding epigenomic pathways that serve as effectors downstream of common mutations in the pancreas, our design seeks to maximize the yield of rapidly translatable mechanistic knowledge. Discoveries from this proposal will have a valuable positive impact because the application of concepts, techniques, and drugs from the field of epigenomics is anticipated to provide new opportunities for the management of patients affected by pancreatic diseases, thereby bearing significant biomedical relevance.