The Novel Approach of RNA Based Therapeutic Technologies

描述（由申请人提供）：本提案的长期目标是设计和开发调节动脉粥样硬化发展和进展的新策略，并为靶向特异性锤头核酶抑制基因表达的应用提供新的见解。我们的最终目标将是开发人类基因治疗策略来缓解动脉粥样硬化，这是冠状动脉疾病和缺血性中风的大多数临床表现的原因。动脉粥样硬化被认为是一种炎症性疾病，LDL存在于内膜（内皮下基质）中，导致炎症反应、氧化应激和斑块形成。我们的实验室开发了一种基因定义的动脉粥样硬化小鼠模型（LDb小鼠），其血浆脂蛋白谱与患有高脂血症的人类相似，并导致饮食无关的动脉粥样硬化。在这个模型中，动脉粥样硬化的病因与人类相似。在这项应用中，我们计划开发新的策略，使用最小和三级稳定的锤头核酶同时靶向两个基因（载脂蛋白B mRNA和脂蛋白相关磷脂酶A2），并将测试该策略对LDb小鼠动脉粥样硬化发展的影响。我们假设，抑制apoB和Lp-PLA2 mRNA的基因表达将减少LDL的氧化，减少炎症反应，并抑制LDb小鼠动脉粥样硬化的发展。我们将使用双链嵌合腺相关病毒载体AAV2/8将这些锤头核酶分子传递给小鼠，并将检测载体和核酶基因在体内表达的持久性。这项研究将使我们了解核酶调节基因表达的分子机制以及这些核酶抑制动脉粥样硬化发展的能力。综上所述，本研究将为冠状动脉粥样硬化提供一种新的治疗方法，并为RNA为基础的分子作为潜在治疗药物的效率和疗效提供证据。

DESCRIPTION (provided by applicant): The long-term objectives of this proposal are to design and develop novel strategies for regulating the development and progression of atherosclerosis and to provide new insights into the application of target specific hammerhead ribozymes for inhibiting gene expression. Our ultimate goal will be to develop human gene therapy strategies for alleviating atherosclerosis, which is responsible for most clinical manifestations of coronary artery disease and ischemic stroke. Atherosclerosis is recognized as an inflammatory disease, with the presence of LDL in the intima (subendothelial matrix) leading to inflammatory responses, oxidative stress, and plaque formation. Our laboratory has developed a genetically defined mouse model of atherosclerosis (LDb mice), which has a plasma lipoprotein profile that resembles humans with hyperlipidemia and results in diet-independent atherosclerosis. The etiology of atherosclerosis in this model mimics that of humans. In this application, we plan to develop novel strategies using minimal and tertiary stabilized hammerhead ribozymes to target two genes (apolipoprotein B mRNA and lipoprotein-associated phospholipase A2) simultaneously and will test the effect of this strategy on atherosclerosis development in LDb mice. We hypothesize that inhibiting gene expression of apoB and Lp-PLA2 mRNA will decrease oxidation of LDL, reduce the inflammatory response, and inhibit atherosclerosis development in LDb mice. We will use the double-stranded chimeric adeno-associated viral vector AAV2/8 to deliver these hammerhead ribozyme molecules to mice and will examine the persistence of both vectors and ribozyme gene expression in vivo. This study will allow us to understand the molecular mechanisms used by ribozymes to regulate gene expression and the ability of these ribozymes to inhibit atherosclerosis development. In summary, this study will provide a novel therapeutic treatment for coronary atherosclerosis and provide evidence of the efficiency and efficacy of RNA based molecules as potential therapeutic agents.