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

基本信息

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
项目类别：
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

项目摘要

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）：将小型干扰RNA（siRNA）引入具有转染试剂的细胞中会导致有效的基因沉默。不幸的是，尽管基于siRNA的功能基因组学在体外广泛使用，但将该技术应用于原代细胞以及体内目标验证的能力受到了有限的有效和无毒输送系统的可用性。我们与RXI Pharmaceuticals合作开发了一类新型的共价RNAi化合物，与传统的siRNA相比，不需要输送工具进入细胞并改善了药理学。我们将这些化合物称为“自递RNA”或SDRNA。 SDRNA是一种疏水的RNAi-Antensens杂种，已证明在原代细胞和局部给药时体内具有高效的体外体外。在几种组织中，已经证明了在包括皮肤，肌肉，肿瘤，肺泡巨噬细胞，脊髓，视网膜等的多种组织中，在细菌层和视网膜中，在5G剂量的SDRNA注射皮层和视网膜上，在5G剂量注射SDRNA的几种组织中已经证明了强烈的摄取和/或沉默。 RXI已利用广泛的筛选来成功识别许多靶标的功能性SDRNA化合物，并正在积极采购其中一些化合物用于临床发育。与常规的siRNA相比，SDRNA作为高功能基因组学工具的商业化是一种优质的功能基因组学工具，它可以在原代细胞和体内启用RNAi。尽管每个基因筛选大量序列的需求并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，在这种基因组中，需要针对数千种基因的具有成本效益的化合物选择。该建议的目的是优化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基于SDRNA的第一行用于商业化的产品创造基础，这将作为第二阶段提案的一部分进行。这些在所有细胞类型中都活跃的SDRNA试剂的可用性，体内将实现功能基因组学和靶向分层/验证研究，并对药物发现过程产生重大影响。公共卫生相关性：RNAi技术的效用受到有限的RNAi化合物向原始细胞和体内有效递送的可用性的严重限制。我们最近开发了一种新的RNAi化合物，即“自交rxRNA”或SDRNA。 SDRNA是一种疏水的RNAi-Antensens杂种，已证明在原代细胞和局部给药时体内具有高效的体外体外。当前的建议将着重于进一步优化这类新的化合物，以实现快速设计和合成。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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