Specific inhibition of transcription factors with Cobalt-Schiff Base Complexes

钴-希夫碱复合物对转录因子的特异性抑制

• 批准号: 9906254
• 负责人: Thomas J Meade
• 金额: $ 29.36万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-15 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9906254
• 关键词: 3-Dimensional; Affinity; Amino Acid Sequence; Attention; BODIPY; Basal cell carcinoma; Behavior; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biology; Breast Cancer cell line; Cancer Biology; Cell Communication; Cell Line; Cells; Chemical Agents; Chemicals; Cobalt; Complex; Consensus Sequence; Coupled; Coupling; Cysteine; DNA; DNA Binding Domain; DNA Sequence; Deuterium; Development; Disease; Doctor of Philosophy; Drosophila genus; Drug Binding Site; Effectiveness; Electron Transport; Embryo; Engineering; Erinaceidae; Family; Family Research; GLI Family Protein; Genetic Transcription; Goals; Growth; Histidine; Homologous Gene; Hydrogen; Impairment; In Situ; Intervention; Ions; Laboratories; Ligands; Light; Malignant Epithelial Cell; Malignant Neoplasms; Mammalian Cell; Medicine; Metals; Modeling; Neoplasm Metastasis; Nonmammalian Embryo; Ocimum basilicum; Oligonucleotides; Oncogenic; Oxidation-Reduction; Pathway interactions; Pattern; Penetration; Process; Prodrugs; Property; Protein Family; Proteins; Regulator Genes; Reporter; Research; Research Project Grants; Role; Schiff Bases; Series; Signal Transduction; Signal Transduction Pathway; Site; Skin; Snails; Specificity; Spectrometry, Mass, Electrospray Ionization; Structure; System; Therapeutic; Therapeutic Agents; Tissues; Topical application; Transition Elements; Universities; Work; Xenopus; Zinc Fingers; base; blocking factor; electron donor; experience; experimental study; in vivo; inhibitor/antagonist; interest; medulloblastoma; metal complex; model development; nanoGold; professor; protein structure; skin disorder; small molecule inhibitor; smoothened signaling pathway; success; therapeutic target; tool; transcription factor; tumor; tumor growth

Project Summary/Abstract The use of metals in medicine has grown impressively in recent years as the result of greatly advanced understanding of the structures of biologically active metal complexes and metal-containing proteins. The goal of this project is to develop a platform that specifically inhibits zinc finger transcription factors (TFs), specifically the Gli family. Gli family TFs represent the final step in the hedgehog (Hh) pathway. The over- activation of this pathway is associated with the growth of a variety of tumors including basal cell carcinoma (BCC) and medulloblastoma. Therefore, inhibitors of these TFs represent potent research tools for biology and have potential use as an entirely new class of therapeutic agents. We are developing cobalt(III)-Schiff base complexes (Co(III)-DNA) that inhibit TFs by a unique mechanism. The complexes are targeted to specific proteins by conjugating decoy oligonucleotides mimicking the native DNA target site of the protein of interest. Subsequent coordination of the cobalt complex to histidines in the zinc finger domain disrupts the structure leading to irreversible inhibition of transcriptional activity. Previous work from our lab has shown that Co(III)-DNA targeted to Snail TFs block EMT in breast cancer cell lines. Preliminary studies in our lab has shown that Co(III)-DNA selectively inhibits Ci (the Drosophila homologue of Gli) in non-mammalian embryo models of development. Here, new Co(III)-DNA conjugates targeted to Gli TFs will be evaluated for their capacity to specifically inhibit Gli transcriptional activity and the Hh pathway in mammalian cell-based reporter assays, and in disease-relevant BCC cell lines. In Aim 1, we will study the interaction of Co(III)-Schiff base and Co(III)-DNA with the Gli DNA binding domain in detail using hydrogen-deuterium exchange coupled with ESI-mass spectrometry. In Aim 2 we will examine the specificity of Co(III)-Gli DNA, comparing targeting to Gli vs. a different transcription factor (Zic) that can bind the same DNA sequence, but with much reduced affinity. In Aim 3, a gold nanoparticle topical delivery system will be used to evaluate the ability of Co(III)-DNA targeted to Gli TFs to disrupt the Hh pathway in whole cell studies and 3D organotypic raft cultures as an epidermal model. We have assembled a team at Northwestern University that includes my research group which has been investigating the physical and biological properties of large number of Co(III)-Schiff base complexes for two decades. Professor Robert Holmgren is a developmental biologist whose research focuses on signal transduction pathways and their role in patterning. Professors Amy Paller, MD and Bethany Perez-White, PhD, whose research focuses on skin cell communication and development. Finally, we have added Professor Anthony Oro (Stanford) who is world leader in Hedgehog signaling pathways and basil cell carcinoma as a consultant (see LOS). The successful completion of this research will produce inhibitors of TFs that function as new research tools and the treatment of a number of diseases that rely on TF signaling.

项目概要/摘要 近年来，由于巨大的影响，金属在医学上的使用取得了令人瞩目的增长。 对生物活性金属配合物和含金属蛋白质结构的深入了解。 该项目的目标是开发一个专门抑制锌指转录因子（TF）的平台， 特别是格利家族。 Gli 家族 TF 代表刺猬 (Hh) 途径的最后一步。过度 该途径的激活与包括基底细胞癌在内的多种肿瘤的生长有关 （BCC）和髓母细胞瘤。因此，这些转录因子的抑制剂代表了生物学和生物医学领域的有效研究工具。 具有作为全新一类治疗剂的潜在用途。我们正在开发钴(III)-希夫碱 复合物（Co(III)-DNA）通过独特的机制抑制 TF。该复合物针对特定 通过缀合模拟目标蛋白质天然 DNA 靶位点的诱饵寡核苷酸来识别蛋白质。 随后钴络合物与锌指结构域中的组氨酸的配位破坏了结构 导致转录活性不可逆的抑制。 我们实验室之前的工作表明，针对 Snail TF 的 Co(III)-DNA 可以阻断乳腺癌中的 EMT 细胞系。我们实验室的初步研究表明，Co(III)-DNA 选择性抑制 Ci（果蝇 Gli的同源物）在非哺乳动物胚胎发育模型中的应用。此处，新的 Co(III)-DNA 缀合物 将评估针对 Gli TF 的特异性抑制 Gli 转录活性的能力以及 基于哺乳动物细胞的报告基因检测以及疾病相关 BCC 细胞系中的 Hh 通路。在目标 1 中，我们 将使用 Gli DNA 结合结构域详细研究 Co(III)-Schiff 碱和 Co(III)-DNA 的相互作用 氢-氘交换与 ESI 质谱联用。在目标 2 中，我们将检查以下方面的特殊性： Co(III)-Gli DNA，比较 Gli 靶向与可结合相同 DNA 的不同转录因子 (Zic) 序列，但亲和力大大降低。在目标 3 中，金纳米颗粒局部给药系统将用于 在全细胞研究和 3D 中评估靶向 Gli TF 的 Co(III)-DNA 破坏 Hh 通路的能力 作为表皮模型的器官型筏培养物。 我们在西北大学组建了一个团队，其中包括我的研究小组，该小组已 研究大量 Co(III)-席夫碱配合物的物理和生物性质 几十年。 Robert Holmgren 教授是一位发育生物学家，其研究重点是信号 转导途径及其在模式形成中的作用。 Amy Paller 教授（医学博士）和 Bethany Perez-White 教授（博士） 他的研究重点是皮肤细胞通讯和发育。最后，我们添加了教授 Anthony Oro（斯坦福大学），Hedgehog 信号通路和罗勒细胞癌领域的世界领导者 顾问（参见 LOS）。这项研究的成功完成将产生 TF 抑制剂，其功能如下： 新的研究工具以及许多依赖 TF 信号传导的疾病的治疗。

项目成果

Cobalt(III) Schiff base complexes stabilize non-fibrillar amyloid-β aggregates with reduced toxicity.

钴(III) 席夫碱复合物可稳定非纤维状淀粉样β 聚集体，并降低毒性。

• 发表时间: 2020
• 影响因子: 3.9
• 作者: Roberts, Kaleigh F;Brue, Christopher R;Preston, Anna;Baxter, Damonick;Herzog, Emma;Varelas, Eleni;Meade, Thomas J
• 通讯作者: Meade, Thomas J