Novel extra cellular RNA-based combinatorial RNA inhibition therapy

基于细胞外RNA的新型组合RNA抑制疗法

• 批准号: 8582255
• 负责人: George A Calin
• 金额: $ 50万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8582255
• 关键词: Affect; Animal Model; Ascites; Biological Assay; Biology; Buffers; Cancer Center; Cancer Patient; Cancer cell line; Categories; Cause of Death; Cell model; Cells; Clinical Trials; Code; Development; Disease; Dose; Drug Kinetics; Drug or chemical Tissue Distribution; Endocrine; Endpoint Determination; Environment; Extracellular Space; Functional RNA; Genes; Goals; Grant; Gynecologic; Hormones; Human; In Vitro; Innovative Therapy; Lead; Libraries; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of ovary; Measurement; Medical; Methods; MicroRNAs; Modeling; Mus; Names; Oncogenic; Organism; Outcome; Ovarian; Ovarian Carcinoma; Pathway interactions; Patients; Pharmacology; Phase; Phenotype; Plasma; Play; Procedures; Production; Protocols documentation; RNA; Research; Research Project Grants; Role; Safety; Sampling; Schedule; Scientist; Serous; Signal Pathway; Signal Transduction; Small Interfering RNA; Small RNA; Technology; Testing; Therapeutic; Time; Tissues; Treatment Efficacy; Tumor Suppressor Proteins; Tumor-Derived; Validation; Vesicle; Work; base; calin; cancer cell; cancer therapy; cancer type; clinical practice; combinatorial; deep sequencing; design; dosage; extracellular; genome wide association study; improved; in vivo; innovation; model design; mortality; nanoliposome; nanoparticle; new technology; novel; novel therapeutic intervention; pre-clinical; preclinical study; public health relevance; safety study; therapeutic development; time use; treatment effect; tumor; uptake

DESCRIPTION (provided by applicant): RNA molecules are secreted in extracellular spaces (exRNAs) and act as endocrine signals altering the phenotypes of cancer cells. In this application we will focus on the class of small non-codingRNAs named microRNAs. We propose an innovative therapeutic concept: the depletion of oncogenic microRNAs from malignant cells by enhancing their secretion as exRNAs or blocking suppressor microRNAs' delivery in extracellular space by blocking secretion mechanisms. Our main goal is to develop new strategies using combined miRNA and siRNA to maximize therapeutic benefit by affecting the production of exRNAs. Our proposal centers on the identification and targeting of specific tumor-derived exRNA and exosomes leading to novel therapies and improved therapeutic outcomes. In UH2 phase of the grant, using ovarian cancer (OC) cell lines and cells derived from OC patient tumors, we will identify novel exRNA-based cancer therapeutic lead candidates by siRNA library genome-wide screening based on secretion measurements, by small RNA deep sequencing in "paired" samples from tumors, plasma and ascites from the same patient, and by functional studies for the lead exRNA candidates. In the UH3 phase, we will achieve pre-clinical optimization by testing the exRNA lead(s) identified in the UH2 for antitumor efficacy in well-established OC murine orthotopic models and their safety using well-established preclinical protocols. The aims in exploratory UH2 phase are designed to identify and validate the function of novel exRNA therapeutic candidates for pre- clinical studies. In UH3 phase, we will formulate the methods for the validation of the assays to determine the activity, pharmacology in tissue and plasma to establish dosage schedules in animal models for the design of rational anti-tumor approaches. Using our nanoliposomal delivery technology, we will also conduct target modulation and efficacy studies to generate clear evidence regarding the safety of exRNA-based therapeutic candidates in the proposed dose range. Our approach, by using a new category of regulatory exRNAs- miRNAs, could substantially enhance therapeutic efficacy for cancer treatment. We will also demonstrate, for the first time, the use of nanoliposome-siRNA-exRNA targeting to generate a synergistic boosting effect for treatment of cancers. While we are focusing on OC due to the high mortality associated with this malignancy, our findings have applications for any type of cancer. This proposal brings synergistic capabilities of scientists working for over a decade in the fields of miRNA (Dr. Calin), siRNA (Dr. Sood), nanoparticle delivery (Dr. Lopez-Berestein), and exosome biology (Dr. O'Halloran). The team has the expertise and synergistic drive to achieve novel exRNA therapeutic development by performing highly innovative research, and has developed RNA-based therapies in the past.

描述(申请人提供)：RNA分子分泌在细胞外间隙(ExRNAs)中，作为内分泌信号改变癌细胞的表型。在本应用程序中，我们将重点介绍一类名为microRNAs的小型非编码RNA。我们提出了一个创新的治疗概念：通过促进肿瘤细胞作为exRNAs的分泌来耗尽肿瘤细胞中的microRNAs，或者通过阻断分泌机制来阻断抑癌microRNAs在细胞外空间的传递。我们的主要目标是开发结合使用miRNA和siRNA的新策略，通过影响exRNAs的产生来最大化治疗效益。我们的建议集中在识别和靶向特定的肿瘤来源的exRNA和exosome，从而导致新的治疗方法和改善的治疗结果。在资助金的UH2阶段，我们将使用卵巢癌(OC)细胞系和OC患者肿瘤细胞，通过基于分泌物测量的siRNA文库全基因组筛选，通过对同一患者的肿瘤、血浆和腹水样本进行小RNA深度测序，以及通过对候选主导exRNA进行功能研究，来确定新的基于exRNA的癌症治疗候选药物。在UH3阶段，我们将使用成熟的临床前方案，通过测试UH2中确定的外源RNA先导(S)在成熟的OC小鼠原位模型中的抗肿瘤效果及其安全性，来实现临床前优化。UH2探索性阶段的目的是识别和验证新的exRNA治疗候选药物的功能，用于临床前研究。在UH3阶段，我们将制定验证方法，以确定活性、组织和血浆中的药理作用，以建立动物模型的给药方案，以设计合理的抗肿瘤方法。利用我们的纳米脂质体递送技术，我们还将进行靶向调节和疗效研究，以产生关于建议剂量范围内基于exRNA的候选治疗药物安全性的明确证据。我们的方法，通过使用一种新的调控exRNAs-miRNAs，可以显著提高癌症治疗的疗效。我们还将首次展示使用纳米脂质体-siRNA-exRNA靶向来产生治疗癌症的协同增强效应。虽然我们专注于OC，因为这种恶性肿瘤的死亡率很高，但我们的发现适用于任何类型的癌症。这项提议带来了十多年来在miRNA(Calin博士)、siRNA(Sood博士)、纳米颗粒递送(Lopez-Berestein博士)和外体生物学(O‘Halloran博士)领域工作的科学家的协同能力。该团队拥有专业知识和协同动力，通过进行高度创新的研究来实现新的exRNA治疗开发，并在过去开发了基于RNA的疗法。