miR-155 and RUNX function in neurofibroma tumorigenesis and therapy

首席调查员/项目主任(最后、第一、中间)：吴建强 使用技术语言的项目摘要，简要描述实现所述目标的研究设计和基本原理 1型神经纤维瘤病(NF1)患者会发展成不同数量的良性雪旺细胞瘤，称为 神经纤维瘤，除手术外没有有效的治疗方法。新的治疗策略和 迫切需要新的目标。我们发现：a)RUNX1和RUNX3的靶向基因同时缺失， 或用RUNX1/CBFβ相互作用抑制剂Ro5-3335药物抑制RUNX功能 小鼠神经纤维瘤细胞在体内增殖和诱导细胞凋亡。B)5个ERK依赖和6个ERK- 与正常小鼠雪旺相比，小鼠神经纤维瘤中独立的MIR被解除调控&>2倍 细胞，c)miR-155受ERK/AP-1调控。D)抗miR-155肽核酸(PNA)抑制小鼠 神经纤维瘤雪旺细胞前体细胞的生长。e)。MIR-155全身击倒部分延迟 神经纤维瘤的形成。我们推测RUNX1、Runx3和miR-155与神经纤维瘤有关 RUNX/CBFβ和/或miR155表达的形成和抑制将为神经纤维瘤的治疗提供新的治疗方法。 我们计划通过使用两种基因工程技术实现以下两个具体目标来检验这一假设 小鼠模型和化学/药物抑制剂。在目标1中，我们将从基因上删除RUNX1和Runx3 同时在雪旺细胞和雪旺细胞前体细胞中应用Cre-loxP技术或灭活RUNX基因 用药物抑制剂抑制Runx/Cbfβ相互作用以确定Runx的功能 神经纤维瘤的形成。我们还将确定RUNX1和RUNX3的调节机制(S) 神经纤维瘤细胞的体外生长。在目标2中，我们将通过遗传方法在体内灭活依赖ERK的miR-155 突变或化学修饰的抗miR-155肽核酸(PNA)，由高水平的 有效的纳米颗粒系统，以确定miR-155在神经纤维瘤形成中的作用。我们将解剖 通过确定NF1缺失如何上调miR-155以及miR-155如何参与MIR-155通路 神经纤维瘤细胞生长。我们将确定miR-155和RUNX1和Runx3之间是否存在因果联系。 总体而言，这一提议将为RUNX1/3和miR-155作为癌基因在 神经纤维瘤的形成，并为临床试验提供临床前的理论基础。

Principal Investigator/Program Director (Last, first, middle): Wu, Jianqiang Project summary Using technical language, briefly describe the research design and rationale for achieving the stated goals Neurofibromatosis type 1 (NF1) patients develop vary number of benign Schwann cell tumors called neurofibromas for which there is no effective therapy available except surgery. New therapeutic strategies and new targets are urgently needed. We showed that: a) targeted genetic deletion of Runx1 & 3 simultaneously, or pharmacological inhibition of Runx function with a Runx1/Cbfβ interaction inhibitor, Ro5-3335, inhibits mouse neurofibroma cell proliferation and induces cell apoptosis in vivo. b) five ERK-dependent and 6 ERK- independent miRs are deregulated >2 folds in mouse neurofibromas compared to normal mouse Schwann cells, c) miR-155 is regulated by ERK/AP-1. d) anti-miR-155 peptide nucleic acids (PNAs) inhibit mouse neurofibroma Schwann cell progenitor growth. e). Whole body knock out of miR-155 partially delays neurofibroma formation. We hypothesize that Runx1 & Runx3 and miR-155 are involved in neurofibroma formation, and inhibition of Runx/Cbfβ and/or miR-155 expression will provide new therapies for neurofibroma. We plan to test this hypothesis by pursuing the following two specific aims using both genetically engineered mouse models and chemical/pharmacological inhibitors. In Aim 1, we will genetically delete Runx1 & Runx3 simultaneously in Schwann cells and Schwann cell precursors by Cre-Loxp technique or inactivate Runx genes by inhibition of Runx/Cbfβ interaction using a pharmacological inhibitor to determine Runx function in neurofibroma formation. We will also determine the mechanism(s) by which Runx1 & Runx3 regulate neurofibroma cell growth in vitro. In Aim 2, we will inactivate ERK-dependent miR-155 in vivo by genetically mutation or chemically modified anti-miR-155 peptide nucleic acids (PNAs) that is delivered by a high efficacious nanoparticle system to determine the role of miR-155 in neurofibroma formation. We will dissect miR-155 pathways by determining how loss of Nf1 upregulates miR-155 and how miR-155 contributes to neurofibroma cell growth. We will determine if there is a causative link between miR-155 and Runx1 & Runx3. Overall, this proposal will provide mechanistic evidence of Runx1/3 and miR-155 function as oncogenes on neurofibroma formation and provide pre-clinical rationale for clinical trials.