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

基本信息

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
项目类别：
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

项目摘要

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）：Hedgehog (Hh) 通路的异常信号传导与基底细胞癌 (BCC) 的形成和进展密切相关，基底细胞癌是一种皮肤癌，影响十分之三的白种人。 BCC 的标准治疗包括手术切除整个肿瘤，但该手术可能会留下严重的疤痕。研究人员一直在研究通过化学抑制 Hh 通路发挥作用的替代疗法。迄今为止，大多数疗法都会抑制 Hh 通路的上游成员 smoothened (Smo)，但该通路随后会产生耐药性。因此，拟议的项目旨在针对 Gli 蛋白，这是一组锌指转录因子 (TF)，充当 Hh 途径的最终步骤。 Meade 教授的实验室开发了一系列钴(III)-希夫碱(Co(III)-sb) 配合物，它们是高度特异性和不可逆的锌指转录因子抑制剂。通过将 Co(III)-sb 与 Gli 靶标结合序列 (5'-GACCACCCA-3') 缀合形成 Co(III)-Gli，预计会对 Gli 蛋白产生高度特异性和有效的抑制作用。该提案还寻求将荧光修饰的 Co(III)-Gli 抑制剂附着到金纳米颗粒 (AuNP) 上，形成“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 可以通过光诱导从 AuNP 中释放，以及 3) 荧光活性的轻松可视化和跟踪体外和体内药剂。该提案的第一个目标是合成和表征独特的 Au-Gli-Co 颗粒。 Co(III)-Gli 将使用 DNA 杂交方法附着到 AuNP。它可以使用近红外光从 AuNP 中脱杂化（并因此释放），从而引起 AuNP 等离激元共振并产生热量。第二个和第三个目标是在体外和体内验证 Hh 通路抑制。细胞实验将在两种细胞系上进行，以确定抑制剂针对 Gli 的功效和特异性。 3D 筏培养模型将用于评估组织穿透和细胞递送。体内实验将确定该药物在局部应用和激光激活后治疗 BCC 肿瘤的能力。将使用在他莫昔芬诱导后表达 Hh 介导的 BCC 肿瘤的小鼠模型。该提案符合 NIAMS 的长期研究和资助计划。该项目涉及开发一种使用小分子的新皮肤癌治疗方法，以及提出一种有效且受控的经皮药物输送系统。它可用于治疗早期或晚期BCC。与许多皮肤病一样，基底细胞癌往往比侵袭性更强的癌症较少受到关注，但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。