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）：耐药性是主要的 征服细菌感染的障碍。该建议针对 细菌传染病治疗的关键问题：需要 制定旨在维护当前可用有效性的策略 氨基糖苷抗生素。调查人员专注于对 由氨基糖苷6'-N-N-乙酰转移酶介导的氨基糖苷氨基甲霉素 AAC（6'） - IB，它也可以修改其他几种氨基糖苷抗生素。这 该研究项目的长期目标是开发反义的使用 寡核苷酸作为选择性抑制AAC表达（6'）的工具 和其他相关基因。该赠款提案的具体目的是：1a） 鉴定促进裂解的合成寡核苷酸和类似物 AAC（6'） - IB mRNA的RNase H或RNase P. 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 可以通过在mRNA中降解对基因表达的抑制作用 存在适当的反义分子。一般策略 具体目的将包括两个概念上不同的利用 识别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 将设计以抑制AAC（6'） - Ib的表达 mRNA的降解。该信息也将用于 反义寡聚核苷酸的发展，抑制表达的表达 AAC（6'） - IB通过诱导mRNA的RNase P降解。以下， 研究人员将开始研究寡核苷酸的细胞摄取和 类似物。 2A）开发抑制的肽核酸（PNA）分子 AAC（6'）-IB的表达。 2b）PNA分子的生物利用度研究。小说 现在可以使用反义技术抑制基因表达的策略 与肽核酸（PNA）的合成。 PNA可以开发为 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 AAC（6'）-IB的表达。然后，调查人员将研究生物利用度 裸露和脂质体封装的PNA分子。