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: 8528523
• 负责人: Flordeliza S Villanueva
• 金额: $ 18.72万
• 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 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8528523
• 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.

描述（由申请人提供）：对肿瘤发生中涉及的途径和蛋白质的了解不断扩大，导致了靶向分子疗法的发展，包括使用设计用作转录因子诱饵的小核酸构建体。尽管有大量基于小核酸的疗法具有潜在的临床价值，但没有可用的方法能够靶向、安全、微创且重复地将治疗量的这些药物递送至癌症。因此，基于小核酸的疗法的临床转化进展缓慢。因此，在本提案中，一个独特的多学科团队，包括化学和生物医学工程、声学物理学、基础和临床肿瘤学、分子生物学和成像方面的专业知识，将开发一种新的转录因子诱饵靶向递送方法，该方法利用超声波诱导的微泡（MB）振动产生的独特生物效应。这些生物效应包括增强细胞膜对大分子的通透性，例如转录因子诱饵，它可以装载在微球上，并在通过指向该部位的超声波诱导微球破裂时仅在目标部位释放。由于超声还具有成像能力，因此项目团队将创新一种治疗诊断超声-MB递送系统，该系统将超声成像与核酸携带能力相结合，共同克服当前治疗性转录因子诱饵递送的障碍，同时允许实时可视化 of the tumor and distribution of the decoy.我们将测试以下总体假设：MB 可以装载治疗性核酸（转录因子诱饵），超声介导的治疗递送将导致癌基因沉默并减少肿瘤生长，并且使用肿瘤特异性靶向部分将增强治疗效果，同时允许特异性肿瘤检测。我们将利用 STAT3 诱饵作为测试治疗剂，因为已知这种小核酸在动物癌症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装载有 STAT3 诱饵的 MB，分析下游靶基因的表达，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递送平台是否在体内诱导STAT3信号沉默和肿瘤生长抑制，我们将静脉内递送负载STAT3诱饵的MB并对患有鳞状细胞癌的小鼠进行超声处理，测定STAT反应基因的表达，并随着时间的推移超声跟踪肿瘤生长。这些研究最终将形成一种有效的、非侵入性的、有针对性的转录因子诱饵递送策略，该策略应有助于临床实施。重要的是，虽然我们提出的递送策略针对该项目中的特定癌基因，但这项工作将为图像引导超声-MB治疗递送平台建立一般原则，该平台可以扩展到以小核酸递送为代表的治疗方法的其他疾病。

项目成果

Suspension-Expansion of Bone Marrow Results in Small Mesenchymal Stem Cells Exhibiting Increased Transpulmonary Passage Following Intravenous Administration.

• DOI: 10.1089/ten.tec.2014.0344
• 发表时间: 2015-02
• 期刊: Tissue engineering. Part C, Methods
• 作者: A. Zanetti;M. Grata;E. Etling;Regeant Panday;F. Villanueva;C. Toma

Radial modulation contrast imaging using a 20-MHz single-element intravascular ultrasound catheter.

使用 20 MHz 单元件血管内超声导管进行径向调制对比成像。

• DOI: 10.1109/tuffc.2014.6805692
• 发表时间: 2014
• 期刊: IEEE transactions on ultrasonics, ferroelectrics, and frequency control
• 作者: Yu,FrancoisTH;Villanueva,FlordelizaS;Chen,Xucai
• 通讯作者: Chen,Xucai

Getting good vibes: the therapeutic power of microbubbles and ultrasound.

• DOI: 10.1016/j.jcmg.2012.09.007
• 发表时间: 2012-12
• 期刊: JACC. Cardiovascular imaging
• 作者: F. Villanueva

Cardiac Gene Expression Knockdown Using Small Inhibitory RNA-Loaded Microbubbles and Ultrasound.

• DOI: 10.1371/journal.pone.0159751
• 发表时间: 2016
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Kopechek JA;Carson AR;McTiernan CF;Chen X;Klein EC;Villanueva FS
• 通讯作者: Villanueva FS

Treatment of microvascular micro-embolization using microbubbles and long-tone-burst ultrasound: an in vivo study.

使用微泡和长色 - 爆破超声处理微血管微栓塞：一项体内研究。

• DOI: 10.1016/j.ultrasmedbio.2014.09.033
• 发表时间: 2015-02
• 期刊: ULTRASOUND IN MEDICINE AND BIOLOGY
• 影响因子: 2.9
• 作者: Pacella, John J.;Brands, Judith;Schnatz, Frederick G.;Black, John J.;Chen, Xucai;Villanueva, Flordeliza S.
• 通讯作者: Villanueva, Flordeliza S.