Project 4

项目4

基本信息

批准号：
10230530
负责人：
Taosheng Chen
金额：
$ 2.56万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-19 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10230530
关键词：
Acute Erythroblastic Leukemia Acute Myelocytic Leukemia Acute leukemia Binding Biological Assay Biophysics C-terminal Cell Nucleus Cell model Cells Cellular Assay Chemicals Child Chromatin Collection DNA Binding Domain Development Disease Epigenetic Process Experimental Models Family Fluorescence Resonance Energy Transfer Fusion Oncogene Proteins Gene Expression Genetic Transcription Goals HOXA9 gene Hematologic Neoplasms Hematopoietic Hematopoietic stem cells Histones Homeobox Human In Vitro Leukemic Cell Libraries Lysine Metabolic Methods Modeling Modernization Mus N-terminal NMR Spectroscopy NUP98 gene Nuclear Nuclear Pore Complex Oncogenes Pediatric Hematology Permeability Pharmaceutical Chemistry Pharmaceutical Preparations Phenotype Pre-Clinical Model Property Protac Regulator Genes Remission Induction Therapy Saint Jude Children&apos s Research Hospital Solubility Structure Surface Testing Therapeutic Transplantation X-Ray Crystallography base biophysical techniques cell transformation chemical synthesis in vivo inhibitor/antagonist leukemia mouse model novel therapeutic intervention outcome forecast prevent recruit screening small molecule small molecule inhibitor submicron therapeutic development transcription factor treatment strategy ubiquitin-protein ligase

项目摘要

Project 4/Summary NUP98-fusion oncogenes (eg, NUP98-HOXA9, -KDM5A & -NSD1) are associated with several pediatric hematological malignancies (eg, AML; 6-10%; and AEL; 20%) characterized by poor prognosis. All of these oncogenes fuse the N-terminal FG-repeat domain of the nucleoporin NUP98 to chromatin binding domains from a variety of transcriptional and epigenetic regulators (e.g., the histone binding PHD3 domain of KDM5A). Endogenous NUP98 primarily localizes to the nuclear pore complex, with its intrinsically disordered FG-repeat domain (~500 residues) filling the central pore. In contrast, NUP98 fusion oncoproteins (FOs) enter the nucleus, bind to chromatin and cause the formation of numerous, sub-micron sized nuclear puncta. Importantly, these puncta recruit epigenetic and transcriptional regulators to developmental regulatory genes, notably the HOX family, activate aberrant gene expression, and transform hematopoietic cells. Leukemias harboring NUP98 FOs are treated with standard AML-based remission induction therapy, with intensification/transplantation for suboptimally responsive disease, but prognosis is generally poor. Therefore, there is great need for new therapeutic strategies for treatment of NUP98 FO-driven leukemias. While the N- terminal domain of NUP98 FOs is considered undruggable because it is intrinsically disordered, their folded C- terminal domains offer opportunities for therapeutic development. For example, binding of the PHD3 domain within the NUP98-KDM5A FO to di-and tri-methylated lysine 4 of Histone 3 (H3K4-Me2/3) is required for rewiring of the epigenetic state of hematopoietic cells and their transformation. Further, deletion of the HOXA9 DNA binding domain from the NUP98-HOXA9 FO alters nuclear puncta formation and prevents cell transformation. Thus, therapeutic strategies targeting the interactions of these folded domains with chromatin have potential to inhibit cell transformation by NUP98 FOs. Here, we propose to develop small molecules that bind the KDM5A PHD3 domain and inhibit its binding to H3K4-Me2/3 (Aims 1 & 2). We further propose to adapt PHD3 domain binders and inhibitors using the PROTAC (PROteolysis-TArgeting Chimaera) strategy to induce degradation of NUP98-KDM5A in cells (Aim 2). The inhibitors and PROTACs we develop will be tested against murine and human cellular models, and in vivo murine models of NUP98-KDM5A-driven AML.

项目4/总结 NUP98-融合癌基因（例如，NUP98-HOXA9、-KDM5A 和 -NSD1）与多种儿科疾病相关血液系统恶性肿瘤（例如，AML；6-10%；和 AEL；20%）的特征是预后不良。所有这些癌基因将核孔蛋白 NUP98 的 N 端 FG 重复结构域融合到染色质结合结构域来自多种转录和表观遗传调节因子（例如 KDM5A 的组蛋白结合 PHD3 结构域）。内源性 NUP98 主要定位于核孔复合体，其 FG 重复序列本质上无序结构域（约 500 个残基）填充中心孔。相比之下，NUP98 融合癌蛋白 (FO) 进入核，与染色质结合并导致形成许多亚微米大小的核斑点。重要的是，这些斑点将表观遗传和转录调节因子招募到发育调节基因中，尤其是 HOX 家族，激活异常基因表达并转化造血细胞。白血病携带 NUP98 FO 的患者接受标准的基于 AML 的缓解诱导疗法，其中针对反应欠佳的疾病进行强化/移植，但预后通常较差。所以，非常需要新的治疗策略来治疗 NUP98 FO 驱动的白血病。而 N- NUP98 FO 的末端结构域被认为是不可成药的，因为它本质上是无序的，它们的折叠 C- 末端结构域为治疗开发提供了机会。例如，PHD3域的结合 NUP98-KDM5A FO 内需要将组蛋白 3 (H3K4-Me2/3) 的二甲基化和三甲基化赖氨酸 4 造血细胞表观遗传状态的重新布线及其转化。此外，HOXA9的删除 NUP98-HOXA9 FO 的 DNA 结合域改变核点形成并阻止细胞转变。因此，针对这些折叠结构域与染色质相互作用的治疗策略具有抑制 NUP98 FO 细胞转化的潜力。在这里，我们建议开发小分子结合 KDM5A PHD3 结构域并抑制其与 H3K4-Me2/3 的结合（目标 1 和 2）。我们进一步建议使用 PROTAC（蛋白水解-目标嵌合体）策略调整 PHD3 结构域结合物和抑制剂诱导细胞中 NUP98-KDM5A 的降解（目标 2）。我们开发的抑制剂和 PROTAC 将接受测试针对小鼠和人类细胞模型，以及 NUP98-KDM5A 驱动的 AML 的体内小鼠模型。