ENHANCING THE INTRACELLULAR FUNCTIONING OF HIV RIBOZYMES

增强 HIV 核酶的细胞内功能

• 批准号: 6170060
• 负责人: John Joseph Rossi
• 金额: $ 29.76万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-03 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6170060
• 关键词: AIDS therapy; Adenoviridae; HIV infections; SCID mouse; antiAIDS agent; cell differentiation; cell free system; disease /disorder model; gene therapy; hematopoietic stem cells; human immunodeficiency virus 1; intracellular transport; method development; ribozymes; tissue /cell culture; transfection; transfection /expression vector

DESCRIPTION: (Adapted from applicant's abstract: Within the past year there have been encouraging results in the treatment of HIV-1 infection resulting from the combinatorial use of drugs targeted against HIV-1 encoded reverse transcriptase and protease. Multiple targeting based upon rational drug design has therefore proven to be an effective method for dramatically inhibiting viral infectivity. Despite the success of the combinatorial drug therapy, there are cautious optimism and concerns about viral resistance, potential toxicity from long term use of the drugs, and long lived viral reservoirs that fully justify further development and testing of other types of antiviral therapies. One form of antiviral therapy that is actively being pursued by a number of laboratories is gene therapy, whereby antiviral genes are permanently introduced into hematopoietic cells to ameliorate viral infectivity. Within the past year, several different gene therapy trials have been initiated. One class of reagents currently in phase 1 clinical trials is ribozymes, which are site-specific RNA cleaving agents. These RNA molecules can be engineered to cleave at hundreds of different sites along the length of the HIV genome, thereby providing a multiple drug targeting strategy. The proposed research extends and takes advantage of lessons learned from our ongoing research efforts with anti-HIV ribozymes. The overall goal of this program is the enhancement of intracellular efficacy of anti-HIV ribozymes by understanding and utilizing intracellular and viral encoded proteins and novel RNA expression strategies to facilitate ribozyme-target co-localization. Concurrent with studies for improving ribozyme efficacy, combinations or ribozyme-expression cassettes will be inserted into Adeno Associated Viral vectors and HIV based lentiviral vectors for transduction into CD34+ hematopoietic progenitor cells, and transfusion into a SCID-hu mouse model. In this system the effects of anti-HIV-1 ribozyme expression on CD34+ cell differentiation and HIV-1 infection will be evaluated. The studies proposed encompass the most comprehensive study of anti-HIV-1 ribozyme function and pre-clinical analyses of ribozymes model to date. The results from this program will greatly facilitate the effective use of ribozyme treatment for HIV-1 infection in a gene therapy setting.

描述：（改编自申请人的摘要：在过去的一年中 在治疗HIV-1感染方面有令人鼓舞的结果 由针对HIV-1编码的药物组合使用的组合使用 逆转录酶和蛋白酶。 基于理性的多个目标 因此，药物设计已被证明是急剧的有效方法 抑制病毒感染。 尽管组合药物成功了 治疗，对病毒抗性有谨慎的乐观和关注， 长期使用药物的潜在毒性，并长期存在病毒 充分证明其他类型的进一步发展和测试的储层 抗病毒疗法。 一种积极的抗病毒疗法形式 受到许多实验室的追求是基因疗法，抗病毒 基因被永久引入造血细胞以改善 病毒感染。 在过去的一年中，几种不同的基因疗法 试验已开始。 目前在第1阶段中的一类试剂 临床试验是核酶，它们是位点特异性的RNA裂解剂。 这些RNA分子可以设计成数百种不同的分子 沿HIV基因组长度的部位，从而提供多种药物 定位策略。 拟议的研究扩展并利用 从我们正在进行的抗HIV核酶的研究工作中汲取了教训。 该计划的总体目标是增强细胞内 通过理解和利用细胞内的抗HIV核酶的功效 和病毒编码的蛋白质和新型的RNA表达策略，以促进 核酶 - 目标共定位。 与改进的研究同时 核酶功效，组合或核酶表达盒将是 插入Adeno相关病毒载体和HIV基慢病毒 转导向CD34+造血祖细胞的向量，并 输入SCID-HU小鼠模型。 在这个系统中 CD34+细胞分化和HIV-1上的抗HIV-1核酶表达 感染将评估。 研究提出的最多 抗HIV-1核酶功能和临床前的综合研究 迄今为止核酶模型的分析。 该程序的结果将 极大地促进有效利用核酶治疗HIV-1 在基因疗法环境中感染。