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（改编自申请人摘要）：耐药性是一种主要的 在征服细菌感染的障碍。这项建议的目标是 治疗细菌感染性疾病的关键问题： 制定战略，以保持现有 氨基糖苷类抗生素研究人员将重点放在对 氨基糖苷类6 '-N-乙酰转移酶介导的氨基糖苷类阿米卡星 AAC（6 '）-Ib也可以修饰其他几种氨基糖苷类抗生素。的 本研究项目的长期目标是开发反义核酸的应用。 寡核苷酸作为选择性抑制aac（6 ′）-Ib表达的工具 和其他相关基因。这项拨款建议的具体目标是：1a） 促进切割的合成寡核苷酸和类似物的鉴定 1b）aac（6 '）-Ib mRNA的生物利用度研究 合成的寡核苷酸和寡核苷酸类似物。RNase H或RNase P 可以通过降解细胞中的mRNA来介导对基因表达的抑制。 存在适当的反义分子。这其中的总体策略 具体目标将包括利用两个概念上不同的 鉴定mRNA中可用于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来抑制aac（6 '）-Ib的表达 mRNA的降解。这些信息还将用于 开发了反义寡核苷酸， aac（6 '）-Ib通过诱导RNA酶P降解mRNA而被抑制。下文 研究人员将开始对寡核苷酸的细胞摄取进行研究， 类似物2a）开发肽核酸（PNA）分子，其抑制 AAC（6 '）-Ib的表达。2b）PNA分子的生物利用度研究。一种新型 现在可以使用反义技术抑制基因表达策略 与肽核酸（PNAs）的合成有关。PNA可以被开发为 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，以干扰 aac（6 ′）-Ib的表达。研究人员将研究生物利用度 裸的和脂质体包封的PNA分子。