Screening for inhibitors of NSD3 as a treatment for lung cancer

筛选 NSD3 抑制剂治疗肺癌

• 批准号: 10562214
• 负责人: Jolanta Grembecka
• 金额: $ 52.95万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10562214
• 关键词: Affinity; Animal Model; Binding; Biochemical; Biological Assay; Biology; Biophysics; Bladder; Breast; Cancer Model; Cancer Patient; Cancer cell line; Cell Line; Cells; Chemicals; Clinical; Clinical Research; Clinical Trials; Complex; Development; Dose; EZH2 gene; Family member; Fluorescein; Fluorescence Polarization; Follicular Lymphoma; Future; Genetic study; Genomics; Goals; Head and neck structure; Histones; In Vitro; Label; Libraries; Link; Lung; Lysine; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Malignant neoplasm of urinary bladder; Mammalian Cell; Measures; Menin; Michigan; Non-Small-Cell Lung Carcinoma; Nuclear Family; Nuclear Receptors; Nuclear Structure; Oncogenes; Oncogenic; Outcome; Pancreas; Patients; Play; Protein Family; Proteins; Reporting; Role; SET Domain; Series; Squamous Cell Lung Carcinoma; Structure-Activity Relationship; Tertiary Protein Structure; Testing; Therapeutic; Transcriptional Activation; Universities; analog; biophysical techniques; cancer cell; drug discovery; druggable target; experimental study; high throughput screening; histone methyltransferase; human disease; in vivo; inhibitor; lung cancer cell; malignant breast neoplasm; member; meter; novel anticancer drug; novel therapeutics; osteosarcoma; overexpression; receptor binding; sarcoma; screening; small molecule; small molecule inhibitor; small molecule libraries; tumorigenesis

Abstract The nuclear receptor binding SET domain protein 3 (NSD3) belongs to the family of NSD histone methyltransferases, which are responsible for mono- and di-methylation of lysine 36 on histone 3 (H3K36me1/2). Increasing number of studies link NSD3 to various cancers, including lung, breast, pancreatic and bladder cancers. Resent study implicates NSD3 as an oncogene in aggressive lung squamous cell carcinoma (LUSC), which represents a sub-type of non-small cell ling cancer with very poor clinical outcome. NSD3 in amplified and/or overexpressed in LUSC, and its role as an oncogene in this sub-type of lung cancer has been well validated using genetic studies, both in vitro and in vivo. Furthermore, the catalytic activity of NSD3 plays a critical role in LUSC oncogenesis, supporting that small molecule inhibitors of the catalytic SET domain of NSD3 could represent new therapeutics for lung cancer patients. However, NSD3 inhibitors have not been reported to date, but are highly desired as new potential therapeutics for patients with LUSC and other cancers with NSD3 amplifications. In this project we propose to develop NSD3 inhibitors by performing high throughput screening (HTS) at CCG, University of Michigan. The HTS will focus on identification of small molecule inhibitors of the catalytic SET domain of NSD3. To date, we developed and validated a fluorescence polarization (FP) assay that will be used as a primary screening assay for HTS to identify NSD3 inhibitors. Small molecules identified by HTS will be validated for binding to NSD3 and for inhibition of its catalytic activity using a series of biochemical and biophysical experiments, including NMR, ITC, and histone methyltransferase assays. Activity of the most potent compounds will be characterized in cell-based studies to assess inhibition of NSD3 in mammalian cells and understand their mechanism of action. Selected NSD3 inhibitors will be tested in a panel of lung cancer cell lines with NSD3 amplifications and/or overexpression for their activity, selectivity, and mechanism of action. In summary, we expect to identify highly valuable chemical probes selectively targeting catalytic SET domain of NSD3, which will be suitable for mechanistic studies in cancer cells. This project will pave the way towards development of more potent NSD3 inhibitors that will be appropriate for in vivo studies in lung cancer models and for future therapeutic implications.

摘要 核受体结合SET结构域蛋白3（NSD 3）属于NSD组蛋白家族 甲基转移酶，其负责组蛋白3上赖氨酸36的单甲基化和二甲基化（H3 K36 me 1/2）。 越来越多的研究将NSD 3与各种癌症联系起来，包括肺癌，乳腺癌，胰腺癌和膀胱癌 癌的最近的研究表明NSD 3是侵袭性肺鳞状细胞癌（LUSC）的癌基因， 其代表了具有非常差的临床结果的非小细胞肺癌的亚型。NSD 3扩增 和/或在LUSC中过表达，并且其作为癌基因在该亚型肺癌中的作用已经被很好地证实。 使用体外和体内的遗传研究进行验证。此外，NSD 3的催化活性起着重要的作用。 在LUSC肿瘤发生中起关键作用，支持NSD 3催化SET结构域的小分子抑制剂 可以代表肺癌患者的新疗法。然而，尚未报道NSD 3抑制剂 但作为LUSC患者和其他NSD 3癌症患者的新的潜在治疗剂是高度期望的。 扩增。在这个项目中，我们建议通过进行高通量筛选来开发NSD 3抑制剂 (HTS)在密歇根大学CCG。HTS将侧重于鉴定小分子抑制剂， NSD 3的催化SET结构域。迄今为止，我们开发并验证了荧光偏振（FP）测定， 将用作HTS的初步筛选试验，以鉴定NSD 3抑制剂。HTS鉴别的小分子 将使用一系列生物化学和生物化学方法验证其与NSD 3的结合和其催化活性的抑制。 生物物理实验，包括NMR、ITC和组蛋白甲基转移酶测定。最有效的活性 化合物将在基于细胞的研究中表征，以评估哺乳动物细胞中NSD 3的抑制， 了解其作用机制。将在一组肺癌细胞系中测试选定的NSD 3抑制剂 与NSD 3扩增和/或过表达的活性、选择性和作用机制有关。在 总之，我们期望鉴定选择性靶向催化SET结构域的高度有价值的化学探针， NSD 3，其将适合于癌细胞中的机制研究。该项目将为实现 开发更有效的NSD 3抑制剂，适用于肺癌模型的体内研究 以及未来的治疗意义。