USP34 as a novel target for treatment of idiopathic pulmonary fibrosis

描述（由申请人提供）：WNT信号的异常过度激活似乎在特发性肺纤维化（IPF）中起主要作用。过度激活的WNT信号抑制剂是IPF治疗的候选药物。USP34是一种新型的去泛素化酶，已被确定为WNT/ -catenin通路中的IPF靶点。它与轴蛋白相互作用，需要去泛素化轴蛋白，使其进入细胞核，在那里增加-连环蛋白信号传导。细胞验证实验表明，USP34通过（-catenin）的核定位积极调节WNT。由于WNT信号在IPF中升高，USP34抑制剂会减少信号传导并具有潜在的治疗作用。Progenra开发了一种新型的异肽酶检测平台，用于测量DUB活性，本研究将利用该检测方法通过高通量筛选发现USP34抑制剂。首先，该分析将配置为384孔板格式的USP34。接下来，它将用于筛选三个小分子文库- ChemDiv蛋白酶文库（2000个化合物），Maybridge HitFinder多样性文库（14,000个化合物）和DiverSet文库（20,000个化合物）-以寻找有效和选择性的USP34抑制剂。将在基于纤维化细胞的WNT信号模型以及细胞毒性和细胞概念验证分析中评估选定的命中点的有效性。在后期阶段，先导化合物将在体内模型（博来霉素治疗小鼠）和药代动力学研究中进行评估。商业目标是一种对IPF有活性的药物制剂。

DESCRIPTION (provided by applicant): Aberrant overactivation of WNT signaling appears to play a major role in Idiopathic Pulmonary Fibrosis (IPF). Inhibitors of overactivated WNT signaling are candidates for therapy of IPF. USP34, a novel de-ubiquitylating enzyme, has been identified as an IPF target in the WNT/(-catenin pathway. It interacts with Axin and is required to de-ubiquitylate Axin, allowing it to go to the nucleus where it increases (-catenin signaling. Cellular validation experiments have demonstrated that USP34 regulates WNT positively via nuclear localization of (-catenin. As WNT signaling is elevated in IPF, a USP34 inhibitor would decrease signaling and have a potentially therapeutic effect. Progenra has developed a novel isopeptidase assay platform for measuring DUB activity, and the work proposed here will utilize this assay to discover inhibitors of USP34 by high throughput screening. First, the assay will be configured for USP34 in a 384 well plate format. Next, it will be employed to screen three small molecule libraries - the ChemDiv Protease Library (2000 compounds), the Maybridge HitFinder diversity library (14,000 compounds), and the DiverSet Library (20,000 compounds) - for potent and selective inhibitors of USP34. The efficacy of selected hits will be evaluated in a fibrosis cell-based model of WNT signaling and in cytotoxicity and cellular proof of concept assays. In later stages, lead compounds will be evaluated in in vivo models (bleomycin treatment of mice) and pharmacokinetics studies. The commercial goal is a pharmaceutical agent active against IPF. PUBLIC HEALTH RELEVANCE: The WNT/(-catenin signaling pathway has been linked to various human diseases, including cancer and Idiopathic Pulmonary Fibrosis (IPF). For these diseases, inhibitors of WNT signaling are potentially useful as therapeutic agents. Progenra proposes to collaborate with academic experts in the WNT pathway and lung fibrosis to discover inhibitors of an enzyme called USP34, which acts to increase WNT signaling in IPF. Inhibitors of USP34 will be identified in high throughput screening using Progenra's assay technology and evaluated for preclinical development as therapeutic agents to treat IPF.