Macrophage Phenotypic Modulators—A Novel Therapeutic Approach to Liver Fibrosis Treatment

项目摘要 肝纤维化/肝硬化是全球第14大死亡原因。尽管这种疾病很流行， 没有治疗肝纤维化/肝硬化的药物。几项具有里程碑意义的临床研究表明， 纤维化甚至肝硬化在患者中是可逆的。在动物模型中，在纤维化、疾病 导致“促纤维化巨噬细胞”分化成“促分解巨噬细胞”。这些巨噬细胞 产生基质金属蛋白酶（MMPs），去除多余的细胞外基质并产生介质 其终止“活化的”肝星状细胞的基质产生， 肝脏本申请中提出的研究旨在利用促纤维化蛋白的表型可塑性。 巨噬细胞和识别化学品/药物，刺激它们转化为亲解决巨噬细胞。 为了实现这一目标，我们已经开发了一种初级筛查方法，该方法利用高含量成像来检测 巨噬细胞表型转变。通过这种测定，我们建议筛选3，000个文库， 用于鉴定药物再利用为巨噬细胞表型的已知作用机制化合物 具有抗纤维化特性的调节剂。我们将筛选一个额外的23,000种化合物的库， 未知的活性，以确定新的抗纤维化。将进一步描述这些屏幕的正面点击率 通过使用Nanostring测定，以中等通量形式，定量几种巨噬细胞表型- 特定的mRNA。来自该二次筛选的最高命中将在生物测定中进一步评价抗- 纤维化活性总的来说，这些研究有可能确定新的抗纤维化药物，不仅限制 进一步的纤维化发展，但通过触发促纤维化蛋白的转化刺激纤维化逆转。 巨噬细胞转化为促分解巨噬细胞。具有这种新活性的化合物的鉴定具有 可能极大地影响肝纤维化/肝硬化的治疗。

Project Summary Liver fibrosis/cirrhosis is the 14th leading cause of death worldwide. Despite the prevalence of this disease, there are no drugs to treat liver fibrosis/cirrhosis. Several landmark clinical studies have demonstrated that fibrosis, and even cirrhosis, are reversible in patients. In animal models, during regression of fibrosis, disease causing, “pro-fibrotic macrophages” differentiate into “pro-resolving macrophages”. These macrophages produce matrix metalloproteinases (MMPs) that remove excess extracellular matrix and produce mediators that terminate matrix production by “activated” hepatic stellate cells, the primary matrix producing cell in the liver. The studies proposed in this application aim to exploit the phenotypic plasticity of pro-fibrotic macrophages and identify chemicals/drugs that stimulate their conversion into pro-resolving macrophages. Towards this goal, we have developed a primary screening assay that utilizes high-content imaging to detect macrophage phenotypic transition in a 384 well format. With this assay, we propose to screen a library of 3,000 compounds of known mechanism of action to identify drugs for repurposing as macrophage phenotypic modulators with anti-fibrotic properties. We will screen an additional library of 23,000 compounds with unknown activities to identify novel anti-fibrotics. Positive hits from these screens will be further characterized by using the Nanostring assay, in a medium throughput format, to quantify several macrophage phenotype- specific mRNAs. Top hits from this secondary screen will be further evaluated in biological assays for anti- fibrotic activity. Collectively, these studies have the potential to identify novel anti-fibrotics that not only limit further fibrosis development but stimulate fibrosis reversal by triggering the conversion of pro-fibrotic macrophages into pro-resolving macrophages. Identification of compounds with this novel activity has the potential to tremendously impact treatment of liver fibrosis/cirrhosis.