Gene therapy for neuronal diseases using rAAV

腺相关病毒（AAV）载体是一种潜在有用的神经基因治疗基因转移载体。利用AAV载体开发新的神经系统疾病治疗策略对基因治疗研究的影响越来越大。我们开发了AAV载体的生产系统，并对其在各种疾病模型中的效用进行了评估。我们开发了一种大规模的方法，利用主动气体系统生产载体，该系统使用大型培养容器在封闭系统中处理人工和经济有效的感染或转染。白细胞介素-10是一种抗炎细胞因子，可能调节动脉粥样硬化疾病的过程。apoe缺陷小鼠肌内注射AAV5-mIL10可通过抗炎和降胆固醇抑制动脉粥样硬化的发生。通过raav介导的RNAi转导，成功地改善了小鼠亨廷顿病相关的神经病理异常，如不溶性蛋白积累和DARPP-32表达下调。HDAC抑制剂的使用将增强raav介导的转导策略在癌症基因治疗中的应用。优越的转导与转导细胞中rAAV串联体的组蛋白相关染色质形式有关。在皮下肿瘤模型的分析中，证实了转基因基因在体内表达的改善和治疗效果。我们开发了基因修饰的间充质干细胞，产生编码HSV-tk的后代载体，旨在进一步提高系统性自杀癌基因治疗的疗效。通过荷瘤裸鼠血管内注射证实了产质间充质干细胞的致瘤性和抗肿瘤作用。

The adeno-associated virus (AAV) vector is a potentially useful gene transfer vehicle for neurologic gene therapies. The development of novel therapeutic strategies for neurological disorder by using the AAV vector has an increasing impact on gene therapy research. We have developed the production system of the AAV vector and evaluated the utility with various disease models.1. We developed a large-scale method to produce vectors with an active gassing system that uses large culture vessels to process labor-and cost-effective infection or transfection in a closed system.2. Interleukin-10 is an anti-inflammatory cytokine that may modulate the atherosclerotic disease process. Intramuscular injection of AAV5-mIL10 into ApoE-deficient mice inhibited atherogenesis through anti-inflammatory and cholesterol-lowering effects.3. Neuropathological abnormalities associated with Huntington disease, such as insoluble protein accumulation and down-regulation of DARPP-32 expression, were successfully ameliorated in mice by the rAAV-mediated RNAi transduction.4. The use of HDAC inhibitor should enhance the utility of rAAV-mediated transduction strategies for cancer gene therapy. The superior transduction was related to the proposed histone-associated chromatin form of the rAAV concatemer in transduced cells. In the analysis with subcutaneous tumor models, improved transgene expression as well as therapeutic effect was confirmed in vivo.5. We developed genetically-modified mesenchymal stem cells that produce progeny vectors encoding HSV-tk, aiming at further augmenting therapeutic efficacy of systemic suicide cancer gene therapy. The tumor tropism and anti-tumor effects of vector-producing MSCs were confirmed by intravascular injection in tumor-bearing nude mice.