Development of aiRNA technology

aiRNA技术的开发

• 批准号: 7911482
• 负责人: Chiang Jia Li
• 金额: $ 48.55万
• 依托单位: BOSTON BIOMEDICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2010-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7911482
• 关键词: 70-kDa Ribosomal Protein S6 Kinases; Algorithms; Attenuated; Boston; Cancer Model; Cells; Chemicals; Colon Carcinoma; Communicable Diseases; Data; Development; Disease; EEF2 gene; Epigenetic Process; Gene Silencing; Gene Targeting; Genes; Genetic; Goals; Growth; Hela Cells; In Vitro; Inflammatory Response; Interferons; Intervention; Length; Mammalian Cell; Mediating; Medicine; Messenger RNA; Modification; Mus; Nucleotides; Phase; Pilot Projects; Process; Property; RNA; RNA Interference; Reporting; Research; Small Business Innovation Research Grant; Specificity; Structure; Technology; Testing; Therapeutic; Tumor Tissue; Variant; Xenograft procedure; base; beta catenin; cost; design; gene function; improved; in vivo; meetings; new technology; novel; phase 1 study; phase 2 study; public health relevance; research study; response; therapy development; tumor; tumor xenograft

DESCRIPTION (provided by applicant): The discovery of RNA interference (RNAi) promises to revolutionize medicine due to its unlimited potential to treat genetic, epigenetic and infectious diseases. RNAi mediated by synthetic short interfering RNA (siRNA) can theoretically be employed to target any disease gene. However, siRNA-mediated silencing is highly variable with many genes in mammalian cells being inefficiently silenced. In addition, siRNA have been reported to activate host interferon-like inflammatory responses, and mediate off-target gene silencing, both of which significantly challenge the application of RNAi in vivo. We have discovered aiRNA (asymmetric interfering RNA), a novel proprietary RNAi technology that is fundamentally different from conventional siRNA and its variations. In our preliminary studies, we have found that aiRNA has superior gene silencing properties (efficacy, potency, duration, onset of action) compared to siRNA in vitro. Moreover, aiRNAi mechanistically abolished sense strand mediated off-target silencing, and significantly reduced interferon responses. In a pilot study, we have also demonstrated that aiRNA directed against beta-catenin can target HT29 colon cancer xenografts in vivo and mediate significant gene silencing. The smaller size and higher potency of aiRNA should significantly reduce the synthesis cost, and may provide an in vivo delivery advantage compared to siRNA. The long-term goal of this SBIR proposal is to optimize aiRNA technology for gene silencing research and for developing RNAi therapeutics. In Phase I we will perform key experiments necessary for the optimization of aiRNA technology. Specifically, we will perform in vitro studies to optimize the design of aiRNA structures (Aim 1). We will examine the specificity and off-target effects of aiRNA-mediated gene silencing and compare them to (Aim 2). We will also determine the gene silencing properties of aiRNA in vivo (Aim 3). Results from these studies are needed to aid the design and development of our subsequent Phase II studies. Our overall goal in Phase II will be to develop and test a clinically compatible targeted therapy for colon cancer using the optimized aiRNA developed in Phase I. For these studies, optimized aiRNA directed against selected colon cancer target genes will be synthesized. The ability of each aiRNA to mediate therapeutic RNAi in vivo will then be assessed using xenograft and syngeneic colon cancer models. Further development of aiRNA may help to unleash the enormous potential of RNAi-based interventions against a wide spectrum of diseases. . PUBLIC HEALTH RELEVANCE: The recent discovery of RNA interference (RNAi), a natural process that allows specific genes to be "switched off" in cells, promises to revolutionize medicine; however, the development of therapies based on synthetic siRNA (short interfering RNA) has met with significant problems including the induction of non-specific effects, synthesis cost, limited efficacy, delivery difficulty in vivo. Recently, we have discovered that aiRNA (asymmetric interfering RNA), a novel proprietary RNAi technology developed at Boston Biomedical Inc., has superior gene silencing properties (efficacy, potency, specificity, duration, onset of action ) compared to siRNA. The research outlined in this proposal is designed to further investigate aiRNA with the long-term goal of optimizing this novel technology for gene silencing research and for developing RNAi therapeutics. .

描述（由申请人提供）： RNA干扰（RNAi）的发现有望彻底改变医学，因为它具有治疗遗传、表观遗传和感染性疾病的无限潜力。由合成的短干扰RNA（siRNA）介导的RNAi理论上可以用于靶向任何疾病基因。然而，siRNA介导的沉默是高度可变的，哺乳动物细胞中的许多基因被无效地沉默。此外，siRNA已被报道激活宿主干扰素样炎症反应，并介导脱靶基因沉默，这两个显著挑战RNAi在体内的应用。 我们发现了aiRNA（不对称干扰RNA），这是一种新颖的专有RNAi技术，与传统的siRNA及其变体有着根本的不同。在我们的初步研究中，我们发现aiRNA在体外与siRNA相比具有上级基因沉默特性（功效、效力、持续时间、作用开始）。此外，aiRNAi机制性地消除了正义链介导的脱靶沉默，并显著降低了干扰素应答。在初步研究中，我们还证明了针对β-连环蛋白的aiRNA可以在体内靶向HT 29结肠癌异种移植物并介导显著的基因沉默。aiRNA的较小尺寸和较高效力应显著降低合成成本，并且与siRNA相比可提供体内递送优势。SBIR提案的长期目标是优化aiRNA技术，用于基因沉默研究和开发RNAi疗法。 在第一阶段，我们将进行优化aiRNA技术所需的关键实验。具体而言，我们将进行体外研究以优化aiRNA结构的设计（目的1）。我们将检查aiRNA介导的基因沉默的特异性和脱靶效应，并将其与（目标2）进行比较。我们还将确定aiRNA在体内的基因沉默特性（目的3）。需要这些研究的结果来帮助我们后续II期研究的设计和开发。 我们在第二阶段的总体目标将是使用在第一阶段开发的优化的aiRNA开发和测试用于结肠癌的临床相容的靶向疗法。对于这些研究，将合成针对所选结肠癌靶基因的优化aiRNA。然后将使用异种移植和同基因结肠癌模型评估每种aiRNA在体内介导治疗性RNAi的能力。aiRNA的进一步开发可能有助于释放基于RNAi的干预措施对广泛疾病的巨大潜力。. 公共卫生关系： RNA干扰（RNA interference，RNAi）是一种能使细胞内特定基因“关闭”的自然过程，它的发现有望给医学带来革命性的变化，然而，基于合成siRNA（short interfering RNA，短干扰RNA）的治疗方法的发展遇到了一些重大问题，包括非特异性效应的诱导、合成成本、疗效有限、体内递送困难等。最近，我们已经发现，aiRNA（不对称干扰RNA），一种由波士顿生物医学公司开发的新型专有RNAi技术，与siRNA相比具有上级基因沉默特性（功效、效力、特异性、持续时间、作用开始）。该提案中概述的研究旨在进一步研究aiRNA，长期目标是优化这种用于基因沉默研究和开发RNAi疗法的新技术。.