Targeted theranostic microbubble vectors for transcription factor decoy delivery

用于转录因子诱饵递送的靶向治疗诊断微泡载体

• 批准号: 8281002
• 负责人: Flordeliza S Villanueva
• 金额: $ 16.58万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-10 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8281002
• 关键词: Acids; Acoustics; Address; Adoption; Animal Cancer Model; Biodistribution; Biological Assay; Biomedical Engineering; Carrying Capacities; Cell Membrane Permeability; Cell membrane; Cells; Chemical Engineering; Clinical; Clinical Oncology; Detection; Development; Diagnostic; Disease; Endocytosis; Foundations; Gases; Gene Silencing; Gene Targeting; Genes; Goals; Growth; Head and neck structure; Image; Imagery; In Vitro; Injection of therapeutic agent; Knowledge; Ligands; Malignant Epithelial Cell; Malignant Neoplasms; Measures; Mediating; Methods; Microbubbles; Modeling; Molecular; Molecular Biology; Molecular Target; Morbidity - disease rate; Mus; Nucleic Acids; Oligonucleotides; Oncogenes; Outcome; Pathway interactions; Patients; Physics; Proteins; Research; Rupture; Safety; Scheme; Signal Transduction; Site; Squamous cell carcinoma; Stat3 protein; System; Techniques; Testing; Therapeutic; Therapeutic Agents; Therapeutic Effect; Time; Translations; Ultrasonography; Western Blotting; Work; base; cancer therapy; design; design and construction; imaging modality; in vivo; innovation; macromolecule; minimally invasive; mortality; multidisciplinary; novel; novel strategies; portability; targeted delivery; theranostics; therapeutic target; transcription factor; tumor; tumor growth; tumorigenesis; user-friendly; vector; vibration

DESCRIPTION (provided by applicant): Expanding knowledge of pathways and proteins involved in oncogenesis has led to the development of targeted molecular therapeutics, including the use of small nucleic acid constructs designed to act as transcription factor decoys. Despite the plethora of small nucleic acid-based therapeutics of potential clinical value, no available method is capable of targeted, safe, minimally invasive, and repeated delivery of therapeutic quantities of these agents to cancers. As a result, clinical translation of small nuclec acid-based therapeutics has been slow. Accordingly, in this proposal, a unique multi-disciplinary team embodying expertise in chemical and biomedical engineering, acoustic physics, basic and clinical oncology, molecular biology, and imaging, will develop a new approach for targeted delivery of transcription factor decoys that capitalizes on unique bioeffects ensuing from ultrasound-induced vibrations of microbubbles (MBs). These bioeffects include enhanced cell membrane permeability to macromolecules, such as transcription factor decoys, which can be loaded on MBs and released solely at the target site upon induction of MB rupture by an ultrasound beam directed at the site. Because ultrasound also confers imaging capability, the project team will innovate a theranostic ultrasound- MB delivery system that combines ultrasound imaging with nucleic acid carrying capacity, which together may overcome current barriers to therapeutic transcription factor decoy delivery, while allowing real time visualization of the tumor and distribution of the decoy. We will test the overall hypothesis that MB can be loaded with a therapeutic nucleic acid (transcription factor decoy), that ultrasound-mediated delivery of the therapeutic will cause oncogene silencing and reduce tumor growth, and that the use of tumor-specific targeting moieties will enhance the therapeutic effect and simultaneously allow specific tumor detection. We will utilize the STAT3 decoy as a test therapeutic, as this small nucleic acid is known to have therapeutic potential and safety in animal cancer models. First, we will expose cultured carcinoma cells to STAT3 decoy- loaded MBs under varying ultrasound conditions, assay the resulting expression of downstream target genes, and determine the ultrasound and MB features which are critical for successful delivery of this particular agent. Then, to determine if the ultrasound-MB delivery platform induces STAT3 signal silencing and tumor growth suppression in vivo, we will intravenously deliver STAT3 decoy-loaded MBs and administer ultrasound to mice bearing squamous cell carcinomas, assay expression of STAT responsive genes, and ultrasonically track tumor growth over time. These studies will culminate in an efficient, non-invasive, targeted transcription factor decoy delivery strategy that should facilitate clinical implementation. Importantly, while our proposed delivery strategy targets a specific oncogene in this project, this work will establish general principles fr an image- guided ultrasound-MB therapeutic delivery platform that can be extended to other diseases for which small nucleic acid delivery represents a therapeutic approach. PUBLIC HEALTH RELEVANCE: Clinical adoption of molecular therapeutics for targeted silencing of abnormal genes involved in diseases such as cancer, including the use of small nucleic acid constructs designed to act as transcription factor decoys, is limited by the lack of strategies for targeted, safe, minimally invasive, and repeated delivery of these constructs. In this proposal, a unique multi-disciplinary team will develop a bedside ultrasound-microbubble diagnostic and delivery system that combines real time ultrasound imaging with transcription factor decoy-loaded microbubbles for ultrasound-induced decoy delivery specifically to tumors. The delivery platform developed in this project should facilitate the clinical translation of approaches using transcription factor decoys for treatment of cancer, ultimately leading to better outcome in patients afflicted with cancer. This platform can also be applied to molecular therapy of other diseases for which targeted gene silencing represents a treatment option.

描述（由申请人提供）：扩大对肿瘤发生的途径和蛋白质的知识导致了靶向分子疗法的发展，包括使用旨在充当转录因子诱饵的小核酸构建体。尽管有大量的潜在临床价值基于小核酸的治疗疗法，但没有可用的方法能够靶向，安全，微创，并且反复递送这些药物的治疗量。结果，基于Nuclec酸的小型疗法的临床翻译速度很慢。因此，在这项建议中，一个独特的多学科团队在化学和生物医学工程，声学物理学，基本和临床肿瘤学，分子生物学和成像方面的专业知识将开发出一种新方法，以靶向转录因子诱导的转录因子诱导，从而利用超声诱导的微振动（Microbubs）的独特生物效果（Mibs）（Mbs）（mbbs）。这些生物效应包括增强对大分子的细胞膜渗透性，例如转录因子诱饵，该诱饵可以在MBS上加载并仅在针对该部位的超声梁诱导MB破裂后仅在目标部位释放。由于超声也赋予了成像能力，因此项目团队将创新一个将超声成像与核酸承载能力相结合的THERANOSTIC超声输送系统，该系统可能会克服当前的治疗转录因子诱饵交付的障碍，同时允许实时可视化 诱饵的肿瘤和分布。我们将测试总体假设，即可以用治疗核酸（转录因子诱饵）加载MB，即超声介导的治疗性递送将导致癌基因沉默并减少肿瘤的生长，并且使用肿瘤特异性靶向部分将增强治疗型效应和同时的特异性thumortanane temultanate。我们将使用STAT3诱饵作为测试治疗，因为已知这种小核酸在动物癌模型中具有治疗潜力和安全性。首先，我们将在不同的超声条件下将培养的癌细胞暴露于STAT3诱饵载荷MBS，测定所得下游靶基因的表达，并确定对于成功递送该特定药物至关重要的超声和MB特征。然后，为了确定超声MB输送平台在体内诱导STAT3信号沉默和肿瘤生长抑制，我们将静脉内输送STAT3诱饵载荷MB，并对具有鳞状细胞癌的小鼠进行超声处理，分析统计反应基因表达的鳞状细胞癌的表达以及超声跟踪肿瘤生长的肿瘤生长。这些研究将最终以有效的，无创的，有针对性的转录因子诱饵输送策略为顶，该策略应促进临床实施。重要的是，尽管我们提出的交付策略针对该项目中的特定癌基因，但这项工作将建立图像引导的超声MB治疗递送平台，可以扩展到其他核酸递送代表治疗方法的其他疾病。 公共卫生相关性：临床采用分子治疗剂，用于针对癌症等疾病的异常基因的靶向沉默，包括使用旨在用作转录因子诱饵的小核酸构建体，受到缺乏针对性，安全，微创和重复传递这些结构的策略的限制。在该提案中，一个独特的多学科团队将开发一个床边超声 - 微泡诊断和递送系统，该诊断和输送系统将实时超声成像与转录因子诱饵诱导的微泡相结合，用于超声诱导的诱饵递送，专门针对肿瘤。该项目中开发的输送平台应促进使用转录因子诱饵治疗癌症的方法的临床翻译，最终导致患有癌症患者的预后更好。该平台也可以应用于其他疾病的分子治疗，该疾病的其他疾病代表了一种治疗选择。