Chemical and genetic studies of RORgamma and its critical roles in IBD

描述(申请人提供)：核激素受体ROR3最近被证明在包括炎症性肠病(IBD)在内的多种疾病的发病机制中发挥关键作用。然而，其监管机制在很大程度上仍不为人所知。我这项提议的目标是识别哺乳动物细胞中调节ROR3活性的基因，并开发出抑制其功能的小分子。这些努力最终可能导致潜在的治疗方法。 用于IBD和各种依赖ROR3的疾病。自从我在纽约大学医学中心的利特曼实验室开始博士后生涯以来，我完成了全基因组RNAi和无偏小分子筛选，使用基于昆虫细胞系的异源ROR3报告系统。从这些筛选中，我在三种不同的果蝇遗传途径和几类具有不同化学模板的拮抗剂中识别出ROR3调节器。我还证实，这些小的复合拮抗剂不仅在果蝇细胞中起作用，而且在老鼠和人类的T细胞中也起作用。在K99期间，我计划通过采用RNA干扰方法来研究小鼠T细胞中已识别的遗传调节因子的哺乳动物同源物(目标1)。对于一个或两个表现出ROR3最特异活性的基因，在R00期间，我将产生条件基因敲除小鼠系，并在IBD小鼠模型上测试它们的效果(目标3)。此外，我还将努力在体内寻找高效的ROR3化学拮抗剂。 K99和早期R00阶段(目标2)。我已经获得了几种化学衍生品作为先导化合物。通过合成和测试各种化学衍生物，我预计我们将能够在体内高效地识别能够抑制ROR3活性的化合物，并在小鼠IBD模型中研究它们的治疗潜力(AIM 3)。我的直接研究目标是通过从遗传和化学上调节核激素受体ROR3的功能来探索IBD的治疗方案，该受体已被证明对某些类型的IBD至关重要。我的长期研究目标是利用这项研究的见解，确定其他类型IBD的遗传靶点及其调节器。

DESCRIPTION (provided by applicant): The nuclear hormone receptor ROR3 was recently shown to play crucial roles in the pathogenesis of multiple diseases including inflammatory bowel disease (IBD). However, its regulatory mechanisms remain largely unknown. My goal of this proposal is to identify genes regulating ROR3 activity in mammalian cells and to develop small molecules that inhibit its function. These efforts may eventually lead to potential therapies for IBD and a variety of ROR3-dependent diseases. Since I started a post-doctoral career at the Littman laboratory at NYU medical center, I completed genome- wide RNAi and unbiased small molecule screens with heterologous ROR3 reporter systems that are based on a insect cell line. From these screens, I identified ROR3 regulators in three distinct Drosophila genetic pathways and several classes of antagonists with distinct chemical templates. I also confirmed that these small compound antagonists function not only in Drosophila cells, but also in mouse and human T cells. During the K99 period, I plan to study mammalian homologues of the identified genetic regulators in mouse T cells by taking RNA interference approaches (Aim 1). For one or two genes exhibiting the most specific activities for ROR3, during the R00 period, I will generate conditional knock-out mouse lines and test their effects on mouse models of IBD (Aim 3). In addition, I will try to identify potent ROR3 chemical antagonists with high in vivo efficacy during the K99 and the early R00 stages (Aim 2). I already acquired several chemical derivatives as lead compounds. By synthesizing and testing various chemical derivatives, I anticipate that we will be able to identify compounds that can inhibit ROR3 activity with high efficiency in vivo and to investigate their therapeutic potentials in the mouse models of IBD (Aim 3). My immediate research goal is to explore therapeutic options of IBD by genetically and chemically modulating the function of the nuclear hormone receptor ROR3, which has been shown to be crucial for certain types of IBD. My long-term research goal is to identify genetic targets and their modulators for additional types of IBD, utilizing insights from this study.