DEALING WITH ANTIBIOTIC RESISTANCE--ANTISENSE TECHNOLOGY

应对抗生素耐药性——反义技术

• 批准号: 6083937
• 负责人: MARCELO E TOLMASKY
• 金额: $ 12.75万
• 依托单位: CALIFORNIA STATE UNIVERSITY FULLERTON
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6083937
• 关键词: acyltransferase; aminoglycoside antibiotics; antisense nucleic acid; drug design /synthesis /production; drug resistance; enzyme activity; gene expression; pharmacokinetics

DESCRIPTION (Adapted from Applicant's Abstract): Drug resistance is a major obstacle in the conquest of bacterial infections. This proposal targets a critical issue in the treatment of bacterial infectious diseases: the need to develop strategies aimed at preserving the effectiveness of currently available aminoglycoside antibiotics. The investigators focus on resistance to the aminoglycoside amikacin mediated by the aminoglycoside 6'-N-acetyltransferase AAC(6')-Ib, which can also modify several other aminoglycoside antibiotics. The long term goal of this research project is to develop the use of antisense oligonucleotides as tools to selectively inhibit the expression of aac(6')-Ib and other related genes. The specific aims for this grant proposal are: 1a) Identification of synthetic oligonucleotides and analogs that promote cleavage of aac(6')-Ib mRNA by RNase H or RNase P. 1b) Bioavailability studies of synthetic oligonucleotides and oligonucleotide analogues. RNase H or RNase P can mediate inhibition of gene expression by degradation of mRNA in the presence of appropriate antisense molecules. The general strategy of this specific aim will consist of the utilization of two conceptually different approaches to identify regions in the mRNA available for interaction with oligonucleotides. With this information, oligodeoxynucleotides and analogues will be designed to inhibit the expression of aac(6')-Ib by inducing RNase H degradation of the mRNA. The information will also be utilized for the development of antisense oligoribonucleotides that inhibit the expression of aac(6')-Ib by inducing RNase P degradation of the mRNA. Following, the investigators will initiate studies on the cell uptake of oligonucleotides and analogues. 2a) Development of peptide nucleic acid (PNA) molecules that inhibit expression of AAC(6')-Ib. 2b) Bioavailability studies of PNA molecules. A novel strategy to inhibit gene expression using antisense technology is now available with the synthesis of peptide nucleic acids (PNAs). PNAs may be developed as antimicrobial agents in prokaryotic systems. The investigators will test the in vitro and in vivo activity of several PNA molecules to interfere with the expression of aac(6')-Ib. The investigators will then study the bioavailability of naked and liposome encapsulated PNA molecules.

描述（改编自申请人的摘要）：耐药性是一个主要的 克服细菌感染的障碍。该提案的目标是 细菌感染性疾病治疗的关键问题：需要 制定旨在保持现有现有措施有效性的战略 氨基糖苷类抗生素。调查人员重点关注抵抗 由氨基糖苷 6'-N-乙酰转移酶介导的氨基糖苷阿米卡星 AAC(6')-Ib，还可以修饰其他几种氨基糖苷类抗生素。这 该研究项目的长期目标是开发反义的用途 寡核苷酸作为选择性抑制 aac(6')-Ib 表达的工具 以及其他相关基因。该拨款提案的具体目标是：1a) 促进裂解的合成寡核苷酸和类似物的鉴定 通过 RNase H 或 RNase P 检测 aac(6')-Ib mRNA。 1b) 生物利用度研究 合成寡核苷酸和寡核苷酸类似物。 RNase H 或 RNase P 可以通过降解 mRNA 来介导基因表达的抑制 适当反义分子的存在。本次的总体策略 具体目标将包括利用两种概念上不同的 识别 mRNA 中可与相互作用的区域的方法 寡核苷酸。有了这些信息，寡脱氧核苷酸和类似物 将被设计为通过诱导 RNase H 抑制 aac(6')-Ib 的表达 mRNA 的降解。该信息还将用于 开发抑制表达的反义寡核糖核苷酸 aac(6')-Ib 通过诱导 RNase P 降解 mRNA。接下来， 研究人员将启动细胞摄取寡核苷酸的研究 类似物。 2a) 开发抑制肽核酸（PNA）分子 AAC(6')-Ib的表达。 2b) PNA 分子的生物利用度研究。一本小说 使用反义技术抑制基因表达的策略现已可用 肽核酸（PNA）的合成。 PNA 可以开发为 原核系统中的抗菌剂。调查人员将测试 几种 PNA 分子的体外和体内活性干扰 aac(6')-Ib的表达。然后研究人员将研究生物利用度 裸露和脂质体封装的 PNA 分子。