Development of algorithm for identification of functional self-delivering RNAi co

功能性自传递RNAi co识别算法的开发

• 批准号: 8124697
• 负责人: Alexey Wolfson
• 金额: $ 22.5万
• 依托单位: ADVANCED RNA TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-10 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8124697
• 关键词: Affect; Algorithms; Alveolar Macrophages; Area; Cell Line; Cells; Central Nervous System Neoplasms; Characteristics; Chemical Structure; Chemicals; Chemistry; Clinical; Collaborations; Complex; Data; Data Set; Dermal; Development; Dose; Foundations; Gene Silencing; Genes; Guanine + Cytosine Composition; Hybrids; In Vitro; Injection of therapeutic agent; Length; Libraries; Linear Regressions; Location; Luciferases; Messenger RNA; Modification; Myomatous neoplasm; Nature; Pharmacologic Substance; Pharmacology; Phase; Positioning Attribute; Process; RNA; RNA Interference; Reagent; Reporter Genes; Research; Retina; Screening procedure; Site; Skin; Small Interfering RNA; Speed; Spinal Cord; Stratification; Structure; System; Tail; Technology; Testing; Therapeutic; TimeLine; Tissues; Toxic effect; Transfection; Validation; Variant; base; cell type; commercialization; cost effective; design; drug development; drug discovery; efficacy testing; functional genomics; improved; in vivo; novel; preference; scaffold; tissue culture; tool; uptake; validation studies

DESCRIPTION (provided by applicant): Introduction of small interfering RNAs (siRNAs) into cells with transfection reagents results in efficient gene silencing. Unfortunately, while siRNA-based functional genomics is widely used in vitro, the ability to apply this technology to primary cells and in vivo target validation has been impeded limited availability of efficient and non-toxic delivery systems. We, in collaboration with RXi Pharmaceuticals, have developed a novel class of covalently modified RNAi compounds that do not require a delivery vehicle to enter cells and have improved pharmacology compared to traditional siRNAs. We term these compounds "self- delivering RNA" or sdRNA. sdRNA is a hydrophobically modified RNAi- antisense hybrid, which has been demonstrated to be highly efficacious in vitro in primary cells and in vivo upon local administration. Robust uptake and/or silencing without toxicity has been demonstrated in several tissues including dermal, muscle, tumors, alveolar macrophages, spinal cord, retina etc. In dermal layer and retina, intradermal and intravitreal injections of sdRNA at 5g doses induce potent and long lasting silencing. RXi has utilized extensive screening to successfully identify functional sdRNA compounds against a number of targets and are actively procuring some of these compounds toward clinical development. A major hurdle to commercialization of sdRNA as a superior functional genomics tool, enabling RNAi in primary cells and in vivo, is the relatively low hit rate as compared to conventional siRNAs. While the need to screen large number of sequences per gene is not a limiting factor for therapeutic applications, it severely limits the applicability of sdRNA technology to functional genomics, where cost effective compound selection against thousands of genes is required. The objective of this proposal is to optimize sdRNA structure, chemistry, targeting position, sequence preferences to develop an algorithm for sdRNA potency prediction. Completion of this project will create a foundation for developing the first line of sdRNAs based products for commercialization, which will be done as part of phase II proposal. Availability of these sdRNA reagents that are active in all cell types and in vivo will enable functional genomics and target stratification/validation studies and have a major impact on the drug discovery process. PUBLIC HEALTH RELEVANCE: The utility of RNAi technology is severely limited by limited availability of efficient delivery of RNAi compounds to primary cells and in vivo. We have recently developed a new class of RNAi compounds, "self- delivering rxRNA" or sdRNA. sdRNA is a hydrophobically modified RNAi- antisense hybrid, which has been demonstrated to be highly efficacious in vitro in primary cells and in vivo upon local administration. Current proposal will focus on further optimizing this new class of compounds to enable rapid design and synthesis.

描述（由申请人提供）：使用转染试剂将小干扰RNA（siRNA）导入细胞中可导致有效的基因沉默。不幸的是，虽然基于siRNA的功能基因组学在体外被广泛使用，但将该技术应用于原代细胞和体内靶向验证的能力受到有效且无毒的递送系统的有限可用性的阻碍。我们与RXi Pharmaceuticals合作开发了一类新型的共价修饰的RNAi化合物，与传统的siRNA相比，它们不需要运载工具就可以进入细胞，并且具有更好的药理学。我们将这些化合物称为“自递送RNA”或sdRNA。sdRNA是疏水修饰的RNAi-反义杂合体，其已被证明在体外在原代细胞中和在体内局部施用时是高度有效的。已经在包括真皮、肌肉、肿瘤、肺泡巨噬细胞、脊髓、视网膜等的几种组织中证明了稳健的摄取和/或沉默而没有毒性。在真皮层和视网膜中，以5g剂量的sdRNA的皮内和玻璃体内注射诱导有效且持久的沉默。RXi已经利用广泛的筛选成功地识别了针对许多靶标的功能性sdRNA化合物，并正在积极采购其中一些化合物用于临床开发。将sdRNA作为上级功能基因组学工具商业化的一个主要障碍，使得能够在原代细胞和体内进行RNAi，是与常规siRNA相比相对较低的命中率。虽然每个基因筛选大量序列的需要不是治疗应用的限制因素，但它严重限制了sdRNA技术对功能基因组学的适用性，其中需要针对数千个基因进行成本有效的化合物选择。本提案的目的是优化sdRNA结构、化学、靶向位置、序列偏好，以开发用于sdRNA效力预测的算法。该项目的完成将为开发第一批基于sdRNA的商业化产品奠定基础，这将作为第二阶段提案的一部分。这些在所有细胞类型和体内都有活性的sdRNA试剂的可用性将使功能基因组学和靶标分层/验证研究成为可能，并对药物发现过程产生重大影响。 公共卫生相关性：RNAi技术的效用受到RNAi化合物有效递送至原代细胞和体内的有限可用性的严重限制。我们最近开发了一类新的RNAi化合物，“自递送rxRNA”或sdRNA。sdRNA是疏水修饰的RNAi-反义杂合体，其已被证明在体外在原代细胞中和在体内局部施用时是高度有效的。目前的提案将集中于进一步优化这类新化合物，以实现快速设计和合成。

项目成果

Functional features defining the efficacy of cholesterol-conjugated, self-deliverable, chemically modified siRNAs.

• DOI: 10.1093/nar/gky745
• 发表时间: 2018-11-16
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Shmushkovich T;Monopoli KR;Homsy D;Leyfer D;Betancur-Boissel M;Khvorova A;Wolfson AD
• 通讯作者: Wolfson AD