Enhancing miRNA Therapeutics through Combinatorial Targeting and Vehicle Free Delivery

通过组合靶向和无载体递送增强 miRNA 治疗

• 批准号: 10237907
• 负责人: Andrea L Kasinski
• 金额: $ 35.46万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10237907
• 关键词: Address; Algorithms; Animal Cancer Model; Antineoplastic Agents; BCL2 gene; Binding; Biodistribution; Biological Assay; Biology; Blood Circulation; Cancer Model; Cells; Clinic; Clinical; Clinical Trials; Collaborations; Complex; Data; Diagnosis; Down-Regulation; Ectopic Expression; Encapsulated; Gene Expression; Generations; Genes; Genetic Transcription; Genome; Growth; Human; Hybrids; Immune response; In Vitro; Individual; KRASG12D; Kinetics; Knowledge; Lead; Libraries; Life; Ligands; Lipids; Lung nodule; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Methods; MicroRNAs; Mission; Modeling; Molecular; Molecular Profiling; Mus; Mutate; Non-Small-Cell Lung Carcinoma; Nucleic Acids; Nucleotides; Oncogenes; Pathway interactions; Patients; Pattern; Phase; Pilot Projects; Public Health; Pulmonary Pathology; RNA; Regulator Genes; Research Personnel; Resolution; Seeds; Signal Pathway; Small RNA; TP53 gene; Target Populations; Testing; Therapeutic; Therapeutic Uses; Time; Tissues; Toxic effect; Transfection; Treatment Efficacy; United States National Institutes of Health; Untranslated RNA; Validation; Work; base; cancer therapy; cell growth; cellular targeting; combinatorial; deep sequencing; design; dosage; gene repression; improved; in vivo; infancy; insight; lead candidate; microRNA replacement therapy; molecular pathology; mouse model; novel; novel therapeutics; phase I trial; targeted delivery; therapeutic miRNA; tumor; tumorigenic; uptake

MicroRNAs (miRNAs) have emerged as powerful regulators of the genome and, through concerted efforts to identify their function and evaluate their ability to alter cell growth in vitro and in vivo, some have gained favor as potential therapeutics. Although these miRNA-based approaches can revolutionize the way that tumors are diagnosed and treated, our understanding of the functional and the molecular aspects of miRNA biology are still incomplete. Moreover, in order to bridge miRNA biology with clinical utility, the challenges that still remain with regard to in vivo delivery of miRNAs must be tackled. To address these challenges we propose two Specific Aims: I. To enhance miRNA therapeutic efficacy through combinatorial miRNA-based targeting and molecular profiling and II. To develop and test second-generation vehicles for delivery of unprotected miRNAs. Our extensive preliminary evidence supports both Aims. We recently identified 10 miRNAs out of 2,019 that significantly enhance the tumor suppressive activity of miR-34a, the first miRNA to enter into clinical trial, and have begun to identify the direct targets of these miRNAs to gain insight into the molecular reason for the cooperative effect. We used a novel method that relies on ligating the cellular miRNAs directly to their associated RNA target followed by deep sequencing of the RNA hybrids. The sequencing data from 13 libraries that we constructed will be used to identify the direct targets of these miRNAs, independent of current algorithms. Targets will be validated and evaluated for pathways that they associate with that will begin to explain their cooperative effect with miR-34a. We also propose to evaluate the in vivo efficacy of the combinatorial pairs using various models of lung cancer. Although we are committed to understanding how these miRNAs are cooperating with miR-34a we also propose to use this data to better understand miRNA biology at a global level. Thus, our data will be used to determine how miRNAs associate with their targets at nucleotide resolution, and how the target population changes with regard to miRNA concentration, which is extremely important to understand as miRNA clinical utility increases. In parallel we will develop and test a second-generation miRNA delivery vehicle, which is a first-in-class method for delivering miRNAs completely unprotected. Following systemic delivery using this method, the miRNA accumulates specifically in the tumor and is efficiently taken up by the tumorigenic cells as indicated by target gene repression with no obvious toxicity. Collectively, the data obtained from this work will validate in vivo efficacy for combinatorial miRNA therapeutics, and for the first time will provide evidence for unprotected miRNA delivery. We will also begin to break down the barriers regarding miRNA target identification that until now has been mostly approached using algorithms that lack critical parameters due to a gap in our understanding of how miRNAs bind their targets.

MicroRNAs(MiRNAs)已经成为基因组的强大调节者，通过共同努力 鉴定它们的功能并评估它们在体外和体内改变细胞生长的能力，一些已经获得了青睐 作为潜在的治疗方法。尽管这些基于miRNA的方法可以彻底改变肿瘤的治疗方式 诊断和治疗，我们对miRNA生物学的功能和分子方面的理解是 仍然不完整。此外，为了将miRNA生物学与临床应用联系起来，仍然存在的挑战 关于体内传递miRNAs的问题，必须加以解决。为了应对这些挑战，我们提出了两项建议 具体目标：一、通过以miRNA为基础的组合靶向和 分子图谱和ii.开发和测试第二代载体，用于交付未受保护的miRNAs。 我们广泛的初步证据支持这两个目标。我们最近在2,019个miRNAs中确定了10个 这大大增强了首个进入临床试验的miRNA miR-34a的抑瘤活性， 并已开始确定这些miRNAs的直接靶点，以深入了解导致 协同效应。我们使用了一种新的方法，依赖于将细胞内的miRNA直接连接到它们的 相关的RNA靶标，然后对RNA杂交物进行深度测序。来自13年的测序数据 我们构建的文库将用于识别这些miRNAs的直接靶标，与当前的 算法。将验证和评估目标所关联的路径，这些路径将开始 解释它们与miR-34a的协同效应。我们还建议评估该疫苗的体内疗效。 使用不同肺癌模型的组合配对。尽管我们致力于了解如何 这些miRNA正在与miR-34a合作，我们还建议使用这些数据来更好地了解miRNA 全球范围内的生物学。因此，我们的数据将被用来确定miRNAs如何与其目标在 核苷酸分辨率，以及目标种群如何相对于miRNA浓度发生变化，这是 随着miRNA临床效用的增加，了解这一点极其重要。同时，我们将开发和测试一个 第二代miRNA递送工具，这是一种一流的完全递送miRNA的方法 没有保护措施。在使用这种方法全身给药后，miRNA在肿瘤中特异地积聚。 如靶基因抑制所示，它被肿瘤细胞有效地摄取，没有明显的 毒性。总的来说，从这项工作中获得的数据将在体内验证组合miRNA的有效性 这将首次提供不受保护的miRNA传递的证据。我们也将开始 打破有关miRNA靶标识别的障碍，到目前为止，这些障碍主要是通过 由于我们对miRNAs如何结合其靶标的理解存在差距，缺乏关键参数的算法。