A Highly Specific, Tissue-Permeable Inhibitor of Gli Transcription Factors

一种高度特异性、组织渗透性的 Gli 转录因子抑制剂

• 批准号: 8718750
• 负责人: Elizabeth Amber Bajema
• 金额: $ 4.27万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-04 至 2018-06-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8718750
• 关键词: A Mouse; Affect; Alkaline Phosphatase; Antineoplastic Agents; Attention; Basal cell carcinoma; Base Sequence; Binding; Biological; Breeding; Caucasians; Caucasoid Race; Cell Line; Cells; Cicatrix; Clinical; Cobalt; Complex; Consensus Sequence; DNA; Development; Dissection; Drosophila genus; Drug Delivery Systems; Effectiveness; Erinaceidae; Evaluation; Excision; FDA approved; Family; Fluorescence; Fluorescence Spectroscopy; Funding; Gold; Heating; Histidine; Homologous Gene; Human; Imagery; In Vitro; Integral Membrane Protein; Lasers; Lead; Left; Light; Link; Luciferases; Malignant Neoplasms; Mammals; Mediating; Membrane; Methods; Modeling; Monitor; Mus; Mutation; National Institute of Arthritis and Musculoskeletal and Skin Diseases; Nucleic acid sequencing; Oligonucleotides; Operative Surgical Procedures; Output; Pathway interactions; Patients; Penetration; Permeability; Pharmaceutical Preparations; Procedures; Protein Binding; Proteins; Publishing; Quality of life; Research; Research Personnel; Resistance; Resistance development; Schiff Bases; Series; Skin; Skin Cancer; Specificity; Staging; Surface; Surface Plasmon Resonance; System; Tamoxifen; Techniques; Testing; Time; Tissues; Topical application; Validation; Zinc Fingers; cancer therapy; design; fluorophore; gel electrophoresis; human SMO protein; improved; in vivo; inhibitor/antagonist; interest; irradiation; locked nucleic acid; medulloblastoma; meetings; member; mouse model; nanoparticle; nuclease; particle; professor; public health relevance; research study; skin disorder; small molecule; smoothened signaling pathway; standard care; success; transcription factor; tumor; A Mouse; Affect; Alkaline Phosphatase; Antineoplastic Agents; Attention; Basal cell carcinoma; Base Sequence; Binding; Biological; Breeding; Caucasians; Caucasoid Race; Cell Line; Cells; Cicatrix; Clinical; Cobalt; Complex; Consensus Sequence; DNA; Development; Dissection; Drosophila genus; Drug Delivery Systems; Effectiveness; Erinaceidae; Evaluation; Excision; FDA approved; Family; Fluorescence; Fluorescence Spectroscopy; Funding; Gold; Heating; Histidine; Homologous Gene; Human; Imagery; In Vitro; Integral Membrane Protein; Lasers; Lead; Left; Light; Link; Luciferases; Malignant Neoplasms; Mammals; Mediating; Membrane; Methods; Modeling; Monitor; Mus; Mutation; National Institute of Arthritis and Musculoskeletal and Skin Diseases; Nucleic acid sequencing; Oligonucleotides; Operative Surgical Procedures; Output; Pathway interactions; Patients; Penetration; Permeability; Pharmaceutical Preparations; Procedures; Protein Binding; Proteins; Publishing; Quality of life; Research; Research Personnel; Resistance; Resistance development; Schiff Bases; Series; Skin; Skin Cancer; Specificity; Staging; Surface; Surface Plasmon Resonance; System; Tamoxifen; Techniques; Testing; Time; Tissues; Topical application; Validation; Zinc Fingers; cancer therapy; design; fluorophore; gel electrophoresis; human SMO protein; improved; in vivo; inhibitor/antagonist; interest; irradiation; locked nucleic acid; medulloblastoma; meetings; member; mouse model; nanoparticle; nuclease; particle; professor; public health relevance; research study; skin disorder; small molecule; smoothened signaling pathway; standard care; success; transcription factor; tumor

DESCRIPTION (provided by applicant): Aberrant signaling of the Hedgehog (Hh) pathway is heavily linked to the formation and progression of basal cell carcinomas (BCC), a skin cancer that affects 3 in 10 Caucasians. The standard treatment of BCC involves surgical excision of the entire tumor, but this procedure can leave heavy scarring. Researchers have been studying alternate therapy options that act by chemically inhibiting the Hh pathway. Most therapies so far inhibit smoothened (Smo), an upstream member of the Hh pathway, but the pathway later develops resistance. Thus, the proposed project aims to target Gli proteins, a set of zinc finger transcription factors (TFs) that act as the terminal step in the Hh pathway. Professor Meade's lab has developed a series of cobalt(III)-Schiff base (Co(III)-sb) complexes that are highly specific and irreversible inhibitors of zinc finger TFs. By conjugating Co(III)-sb to the Gli targe binding sequence (5'-GACCACCCA-3') to make Co(III)-Gli, highly specific and potent inhibition of Gli proteins is anticipated. This proposal additionally seeks to attach fluorescently modified Co(III)-Gli inhibitor to a gold nanoparticle (AuNP), making "Au-Gli-Co." This will allow for: 1) topical delivery of the Gli inhibitor, 2) localized delivery, since Co(III)-Gli can be released fro the AuNP by inducing with light, and 3) facile visualization and tracking of the fluorescent active agent in vitro and in vivo. The first objective of the proposal is to synthesize and characterize the unique Au-Gli-Co particle. Co(III)-Gli will be attached to the AuNP using DNA hybridization methods. It can become dehybridized (and therefore released) from the AuNP using near-infrared light, which causes AuNP plasmon resonance and generates heat. The second and third objectives are to validate Hh pathway inhibition in vitro and in vivo. Cellular experiments will be performed on two cells lines to determine the inhibitor's efficacy and specificity for targeting Gli. A 3D raft cultre model will be used to assess tissue penetration and cellular delivery. In vivo experiments will determine the ability of the agent to treat BCC tumors after topical application and laser activation. A mouse model that expresses Hh-mediated BCC tumors upon induction with tamoxifen will be used. This proposal meets the long-range research and funding plans of the NIAMS. The project involves the development of a new skin cancer treatment with the use of small molecules, in addition to proposing an efficient and controlled system of transcutaneous drug delivery. It could have application in treating early or later stage BCC. Like many skin diseases, BCC tends to garner less attention than more aggressive cancers, but the successful implementation of this project would have significant implications in improving patients' quality of life.

描述（由申请人提供）：刺猬（HH）途径的异常信号与基底细胞癌（BCC）的形成和进展密切相关，这是一种影响10种高加索人中3分之3的皮肤癌。 BCC的标准治疗方法涉及整个肿瘤的手术切除，但此过程可能会留下重度疤痕。研究人员一直在研究通过化学抑制HH途径起作用的替代治疗选择。到目前为止，大多数疗法都抑制了HH途径上游成员的平滑疗法（SMO），但该途径后来发展出抗性。因此，拟议的项目旨在靶向GLI蛋白，Gli蛋白是一组锌指转录因子（TFS），它们是HH途径中末端步骤的一组。 Meade教授的实验室已经开发了一系列钴（III）-schiff基地（CO（III）-SB）配合物，它们是锌指TFS的高度特异性且不可逆的抑制剂。通过将CO（III）-SB结合到GLI TARGE结合序列（5'-GACCACCCA-3'）以使CO（III）-GLI，高度特异性和有效的GLI蛋白抑制。该提案还试图将荧光修饰的CO（III）-GLI抑制剂连接到金纳米颗粒（AUNP）上，制成“ Au-Gli-Co”。这将允许：1）GLI抑制剂的局部递送，2）局部递送，因为CO（iii）-GLI可以通过诱导光线来释放AUNP，而3）荧光活动的便捷可视化和跟踪 体外和体内的代理。 该提案的第一个目的是合成和表征唯一的Au-Gli-CO粒子。 CO（iii）-GLI将使用DNA杂交方法连接到AUNP。它可以使用近红外光线从AUNP脱生（并因此释放），从而引起AuNP等离子体共振并产生热量。第二和第三目标是验证体外和体内抑制HH途径。细胞实验将在两个细胞系上进行，以确定靶向GLI的抑制剂功效和特异性。将使用3D筏模型来评估组织穿透和细胞递送。体内实验将确定局部应用和激活后药物治疗BCC肿瘤的能力。将使用他莫昔芬诱导后表达HH介导的BCC肿瘤的小鼠模型。 该建议符合NIAMS的远程研究和资金计划。该项目涉及通过使用小分子的新皮肤癌处理的发展，此外还提出了有效且受控的经皮药物递送系统。它可以在早期或晚期BCC治疗时应用。像许多皮肤疾病一样，BCC往往比更多的侵略性癌症吸引注意力更少，但是该项目的成功实施将对改善患者的生活质量产生重大影响。