Development of gene transfer approaches for neuroAIDS

描述(由申请人提供)： 人类免疫缺陷病毒(HIV)是一种原发的中枢神经系统(CNS)疾病，约20%的HIV感染者受到影响。HIVD的治疗是有限的，部分原因是许多抗逆转录病毒药物未能穿透血脑屏障；因此需要新的方法。一种这样的方法可能是以正常情况下通过血脑屏障的细胞为目标--例如单核细胞。这一提议的假设是，有可能对单核细胞进行基因改造，并将其用作“特洛伊木马”递送系统，将基因转移到中枢神经系统，用于治疗神经艾滋病。 该项目将在夏威夷大学和罗切斯特大学的研究人员之间进行综合、协调的合作。该项目的申请部分将侧重于测试和比较分析不同病毒载体转导血单核细胞的潜在能力；还将评估载体在单核细胞跨血脑屏障迁移方面的努力。还将进行实验，以确定(1)转导有缺陷的干扰慢病毒载体(DLV)的单核细胞是否较不允许复制传染性HIV-1，以及(2)是否可以动员、扩增DLV并通过传染性HIV-1传播到未转导的旁观者细胞。这项应用的合作者部分将专注于分析载体转导的单核细胞是否释放与细胞激活相关的可溶性神经毒性因子。此外，该合作者还构建了编码可溶性神经保护因子的病毒载体，并将确定用该载体转导的单核细胞是否能够促进暴露于明确定义的候选HIV神经毒素的旁观者神经元的存活。整个项目是一个紧密结合的整体，将使调查人员的互补专业知识能够以最大限度的生产力和互惠互利的方式得到应用。合作还将为该协会及其学生和研究员提供充分的机会，以获得神经科学和广泛的相关技术方面的培训。总体而言，这项工作有望带来对治疗神经艾滋病和其他神经疾病的新方法的重大洞察。

DESCRIPTION (provided by applicant): Human immunodeficiency virus (HIV) is a primary disorder of the central nervous system (CNS) that affects about 20% of individuals infected with HIV. Treatment for HIVD are limited, in part because many antiretroviral drugs fail to penetrate the blood-brain barrier (BBB); novel approaches are therefore needed. One such approach may be to target cells that normally traffic across the BBB- such as blood monocytes. The hypothesis of this proposal is that it may be possible to genetically modify bolld monocytes and to use them as a "Trojan horse" delivery system, to effect gene transfer into the CNS, for treatment of neuroAIDS. This project will be conducted as an integrated, coordinated collaboration between investigators at the University of Hawaii and the University of Rochester. The applicant component of this project will focus on the testing and comparative analysis of different virus vectors for their potential ability to transduce blood monocytes; vectors will also be evaluated with respect to their efforts on monocyte transmigration across the blood-brain barrier. Experiments will also be conducted to determine (1) whether monocytes transduced with a defective interfering lentivirus vector (DLV) are less permissive for replication of infectious HIV-1, and (2) whether the DLV can be mobilized, amplified and spread to untransduced bystander cells by infectious HIV-1. The collaborator component of this application will focus on analyzing whether vector-transduced monocytes release soluble neurotoxic factors associated with cellular activation. In addition, the collaborator with construct a virus vector encoding a soluble neuroprotective factor, and will determine if monocytes transduced with this vector are capable of promoting the survival of bystander neurons exposed to well-defined candidate HIV- neurotoxins. The entire project comprises a tightly integrated whole, and will allow for the complementary expertise of the investigators to be applied in a maximally productive and mutually beneficial way. The collaboration will also provide ample opportunitites for the PI, and his students and fellows, to obtain training in neuroscience and a wide range of related techniques. Overall, the work is expected to result in significant insight into new approaches for treatment of neuroAIDS and other neurologic diseases.