Project 4

项目4

• 批准号: 10228889
• 负责人: Taosheng Chen
• 金额: $ 2.28万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 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/10228889
• 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' 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

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/摘要 NUP98-融合癌基因(例如，NUP98-HOXA9、-KDM5A和-NSD1)与多种儿科疾病相关 恶性血液病(如6-10%的AML和20%的AEL)预后不良。所有这些都是 癌基因将核孔蛋白NUP98的N端FG重复结构域与染色质结合区融合 来自各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，其固有的无序的FG重复序列 结构域(~500个残基)填充中央孔隙。相比之下，NUP98融合癌蛋白(FOS)进入 细胞核，与染色质结合，形成许多亚微米大小的核斑点。 重要的是，这些斑点招募了发育调控基因的表观遗传和转录调控因子， 尤其是HOX家族，激活异常基因表达，转化造血细胞。白血病 携带NUP98 FOS的患者接受基于AML的标准缓解诱导治疗， 强化/移植治疗反应不佳的疾病，但预后一般较差。因此， 对于NUP98FO驱动的白血病的治疗，迫切需要新的治疗策略。而N- NUP98 FOS的末端结构域被认为是不可药物的，因为它本质上是无序的，它们折叠的C- 终端域为治疗发展提供了机会。例如，PHD3结构域的结合 在NUP98-KDM5A中，组蛋白3的二甲基化和三甲基化赖氨酸4(H3K4-Me2/3)需要 重组造血细胞的表观遗传状态及其转化。此外，删除HOXA9 NUP98-HOXA9 FO的DNA结合域改变核斑点的形成并阻止细胞 转型。因此，针对这些折叠结构域与染色质相互作用的治疗策略 有可能抑制NUP98-FOS的细胞转化。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(蛋白质降解-靶向嵌合体)策略使PHD3结构域结合和抑制物适应于 诱导NUP98-KDM5A在细胞中的降解(目标2)。我们开发的抑制剂和PROTAC将进行测试 抗小鼠和人细胞模型，以及NUP98-KDM5A驱动的AML小鼠体内模型。