Discovery of UHRF1 inhibitors in the treatment of hepatocellular carcinoma

项目总结(试点项目1) 这一合作试点提案的首要目标是开发一种新的治疗方法 肝细胞癌(肝细胞癌)-一种癌症，在美国发病率较高，影响的人口比例较低。 肝细胞癌的标志之一是全球DNA低甲基化水平的表现，这有助于 肿瘤的进展和转移。尽管表观遗传治疗方法正在上升，但有 目前还没有药物能从机制上减少DNA的低甲基化。朝着治疗的目标前进 我们将开发降低DNA含量的化合物 通过一种新的治疗靶点实现低甲基化。具有植物和同源结构域的泛素样蛋白(PHD)和 真正有趣的新基因(环)指域1(Uhrf1)是DNA甲基组的主要调节者。 在肝细胞癌和肿瘤进展中，uhrf1的过表达导致DNA低甲基化。Uhrf1是E3连接酶 通过泛素化影响从头DNA甲基转移酶DNMT3A的负调控，并 退化。DNMT3A的缺失反过来会导致癌症中的整体低甲基化。因此，作为一种手段 为了减少肝细胞癌的全球低甲基化，这一试点项目的重点是开发小甲基化- 抑制uhrf1-DNMT3A蛋白质-蛋白质相互作用(PPI)的分子或类肽。一种化合物 抑制这种相互作用将挽救肝癌细胞中的DNMT3A水平，导致基因组减少 低甲基化与抑制肝细胞癌增殖。以确定具有uhrf1功能的化合物- DNMT3A抑制剂，我们在特定目标1中提出以并行方式部署两个发现平台：DNA- 编码化学技术(DEC-Tec)和珠状类肽筛选。这些平台中的每个都提供 以经济的方式进入一个很大的化学空间，很可能需要这个空间来鉴定能够 干扰uhrf1-DNMT3A相互作用。在特定目标2的支持下，我们将对生物 Uhrf1-DNMT3A抑制剂对癌细胞的作用 来自非洲裔美国人和西班牙裔/拉丁裔患者的肝癌肿瘤。这些都非常有用 癌细胞株将使我们能够在大多数人群的遗传背景下开发我们的化合物 容易患上肝细胞癌。我们将确定低甲基化还原化合物对细胞DNA的影响 甲基化水平，基因表达，以及癌细胞的增殖和凋亡。药物化学将成为 进行以提高化合物效力和生物功效，从而产生大约六种最终化合物 我们将对其进行初步的药理研究。这一项目的成功完成将 为肝癌的治疗提供了一种新的治疗机制，并在 极大地影响了UR种群。

PROJECT SUMMARY (PILOT PROJECT 1) The overarching goal of this collaborative pilot proposal is the development of a novel approach to treat hepatocellular carcinoma (HCC)—a cancer affecting underrepresented populations in the US at a higher rate. One of the signatures of HCC is the manifestation of global levels of DNA hypomethylation which contributes to tumor progression and metastasis. Although epigenetic therapeutic approaches are on the rise, there are currently no drugs that mechanistically function to reduce DNA hypomethylation. Toward the goal of treating UR populations that are significantly impacted by HCC, we will develop compounds that reduce DNA hypomethylation through a novel therapeutic target. Ubiquitin-like with plant and homeodomain (PHD) and really interesting new gene (RING) finger domains 1 (UHRF1) is a master regulator of the DNA methylome. Overexpression of UHRF1 drives DNA hypomethylation in HCC and tumor progression. UHRF1 is an E3 ligase that affects the negative regulation of the de novo DNA methyltransferase DNMT3A through ubiquitination and degradation. Loss of DNMT3A, in turn, leads to global hypomethylation in cancers. Therefore, as a means toward reducing global hypomethylation in HCC, the focus of this pilot project is the development small- molecules or peptoids that inhibit the UHRF1-DNMT3A protein-protein interaction (PPI). Compounds that inhibit this interaction will rescue DNMT3A levels in HCC cells leading to decreases in genomic hypomethylation and the mitigation of HCC proliferation. To identify compounds that function as UHRF1- DNMT3A inhibitors, we propose in Specific Aim 1 to deploy two discovery platforms in a parallel fashion: DNA- Encoded Chemistry Technology (DEC-Tec) and on-bead peptoid screening. Each of these platforms provides an economical access to a large chemical space that will likely be needed to identify compounds capable of disrupting the UHRF1-DNMT3A interaction. Under the aegis of Specific Aim 2, we will evaluate the biological effects of the UHRF1-DNMT3A inhibitors by executing a suite of biological assays on cancer cell lines generated from HCC tumors obtained from African American and Hispanic/Latino patients. These highly useful cancer cell lines will allow us to develop our compounds within the genetic backgrounds of populations most susceptible to HCC. We will determine the effects of hypomethylation reducing compounds on cellular DNA methylation levels, gene expression, and cancer cell proliferation and apoptosis. Medicinal chemistry will be performed to improve compound potency and biological efficacy leading to approximately six final compounds for which we will conduct preliminary pharmacological studies. The successful completion of this project will afford a novel therapeutic mechanism for the treatment of HCC and have particularly salutary relevance within greatly affected UR populations.