Developing Nucleic Acid Therapeutics

开发核酸疗法

基本信息

批准号：
7227454
负责人：
Alan M Gewirtz
金额：
$ 26.75万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-08-01 至 2008-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7227454
关键词：
Address Advanced Development Affinity Animal Model Biological Models Cells Chemistry Cleaved cell Clinical Clinical Research DNA Data Defective spinal cord development Development Diagnostic Dose Drug Administration Schedule Drug Delivery Systems Drug Kinetics Dysmyelopoietic Syndromes Engineering Fluorescence Funding Gene Expression Gene Silencing Gene Targeting Genes Goals Guanosine Monophosphate Human In Vitro Laboratory Study Lead Leukemic Cell Life Liquid substance Location MYB gene Malignant Neoplasms Maps Mass Chromatography Measures Messenger RNA Methods Modification Nucleic Acids Organ Patients Pharmaceutical Preparations Pharmacodynamics Phase Phase I Clinical Trials Property Proteins Proto-Oncogene Proteins c-myb RNA RNA Folding Rapid Access to Intervention Development Refractory Reproduction spores Research Design Research Personnel Signal Transduction Site Spectrometry System Techniques Testing Therapeutic Time Tissue Banks Toxic effect Treatment Protocols Vertebral column antisense nucleic acid base cancer cell cell behavior cell killing clinical efficacy design experience improved in vivo killings leukemia mRNA Expression neoplastic cell phosphorothioate programs self quenching reporter molecule uptake

项目摘要

DESCRIPTION (provided by applicant): The prospect of employing antisense nucleic acids (ASNA) for treating human malignancies remains tantalizing, but unrealized. We hypothesize that ASNA would make better drugs if fundamental problems related to mRNA target sequence selection, and molecule delivery, were solved. The goal of this project is to address these core issues in the following specific aims: Aim #1- Develop A Rational Method For Targeting ASNA- We have developed self-quenching reporter molecules (SQRM) that signal only after hybridization, and have used these probes to "map" hybridization accessible sites in mRNA. We will determine the in vivo utility of this strategy by mapping additional target mRNAs, and measuring the efficiency with which sequence directed ASNA inhibit gene expression in living cells. The hypothesis that naturally occurring intracellular proteins might enhance ASNA/mRNA hybridization will also be tested. Candidate proteins will be identified by function, and by using affinity chromatography, and mass spectrometry. Finally, the utility of DNA backbones with enhanced strand invasion properties, with or without proteins that facilitate hybridization, will also be explored; Aim #2- Test the Hypothesis that Gene Silencing Efficiency of Rationally Targeted ASNA Can be Enhanced by Backbone, Sequence, or Pendant Modifications- ASNA cleave mRNA by enzymatic activity engineered into the molecule, or by activating endogenous RNaseH. We will examine the ability of rationally targeted ASNA, synthesized with various backbone modifications, to cleave mRNA targets in vitro and in vivo. The ability of SQRM with pendants that can be photoactivated to detect, and kill, cells on the basis of target mRNA expression will also be examined; Aim #3. Examine the ability of rationally designed, activity optimized ASNA to detect, and kill, tumor cells in animal models of human leukemia- Pharmacodynamic studies of optimized molecules will be undertaken, and their ability to serve as diagnostic, and therapeutic molecules in animal models of human leukemia will be explored.

描述（由申请人提供）：使用反义核酸（ASNA）治疗人类恶性肿瘤的前景仍然诱人，但未实现。我们假设如果解决与mRNA靶序列选择和分子递送有关的基本问题，ASNA将产生更好的药物。该项目的目的是在以下具体目的中解决这些核心问题：目标＃1-开发一种靶向ASNA-的理性方法 - 我们已经开发了自Quench的记者分子（SQRM），仅在混合杂交后发出信号，并将这些探针用于“ MAP” MAP“ MAP” MRNA中的杂交位点。我们将通过绘制其他靶标mRNA，并测量序列定向asna抑制活细胞中基因表达的效率来确定该策略的体内效用。天然存在的细胞内蛋白可能会增强ASNA/mRNA杂交的假设。候选蛋白将通过功能和使用亲和色谱法和质谱法鉴定。最后，还将探索具有增强的链浸入特性的DNA骨架的效用，或者还将探索具有促进杂交的蛋白质的有或没有蛋白质的效用。目的＃2-检验以下假设：可以通过骨链，序列或吊坠修饰来增强基因沉默效率 - 通过酶促活性通过酶促的酶促活性或激活内源性RNASEH来提高肌裂解mRNA。我们将研究以各种主链修饰合成的理性靶向asna的能力，可以在体外和体内裂解mRNA靶标。也将研究基于目标mRNA表达的SQRM与可以光活化检测和杀死细胞的吊坠的能力。目标＃3。将探索，将探索针对优化分子的人类白血病模型中的合理设计，优化ASNA检测和杀死肿瘤细胞的能力，并将探索它们作为诊断性分子的能力，并探索它们在人类白血病动物模型中的治疗分子的能力。