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.

抽象的 核受体结合 SET 结构域蛋白 3 (NSD3) 属于 NSD 组蛋白家族 甲基转移酶，负责组蛋白 3 (H3K36me1/2) 上赖氨酸 36 的单甲基化和二甲基化。 越来越多的研究将 NSD3 与多种癌症联系起来，包括肺癌、乳腺癌、胰腺癌和膀胱癌 癌症。最近的研究表明 NSD3 是侵袭性肺鳞状细胞癌 (LUSC) 的癌基因， 它代表非小细胞癌的一种亚型，临床结果非常差。扩增的 NSD3 和/或在 LUSC 中过度表达，其作为癌基因在这种肺癌亚型中的作用已得到充分证实 使用体外和体内遗传学研究进行验证。此外，NSD3的催化活性还发挥着 在 LUSC 肿瘤发生中发挥关键作用，支持 NSD3 催化 SET 结构域的小分子抑制剂 可能代表肺癌患者的新疗法。然而，NSD3抑制剂尚未见报道。 日期，但非常需要作为 LUSC 和其他 NSD3 癌症患者的新潜在疗法 放大。在这个项目中，我们建议通过进行高通量筛选来开发 NSD3 抑制剂 (HTS)，CCG，密歇根大学。 HTS 将重点鉴定小分子抑制剂 NSD3 的催化 SET 结构域。迄今为止，我们开发并验证了一种荧光偏振 (FP) 检测方法 将用作 HTS 的初步筛选分析，以鉴定 NSD3 抑制剂。 HTS 鉴定的小分子 将使用一系列生物化学和方法验证与 NSD3 的结合及其催化活性的抑制 生物物理实验，包括 NMR、ITC 和组蛋白甲基转移酶测定。最有力的活动 化合物将在基于细胞的研究中进行表征，以评估哺乳动物细胞中 NSD3 的抑制作用， 了解它们的作用机制。选定的 NSD3 抑制剂将在一组肺癌细胞系中进行测试 具有 NSD3 扩增和/或过表达的活性、选择性和作用机制。在 总之，我们期望识别出选择性靶向催化 SET 结构域的非常有价值的化学探针 NSD3，适用于癌细胞的机制研究。该项目将为 开发更有效的 NSD3 抑制剂，适用于肺癌模型的体内研究 以及未来的治疗意义。