Gene Therapy of AIDS Dementia

描述（由申请方提供）：人类免疫缺陷病毒I型（HIV-1）感染可导致某些HIV-1感染个体中枢神经系统（CNS）出现一系列破坏性临床状况。艾滋病痴呆综合症（ADC）是一个统称，用于描述与艾滋病相关的认知功能障碍、运动困难、协调异常和其他神经系统体征和症状。血脑屏障主要由微血管内皮细胞（MVECs）构成，是常规抗逆转录病毒药物进入脑组织的潜在障碍。基因治疗是治疗神经退行性疾病，特别是艾滋病痴呆的最有前途的途径之一。我们的实验室在最近的过去已经利用逆转录病毒载体在CNS为基础的细胞在体外和体内的基因传递。将细胞类型特异性基因递送到中枢神经系统的不同细胞中将是成功的人类神经系统疾病基因治疗的主要先决条件之一。在过去的十年中，多恩堡博士的实验室在构建细胞类型特异性逆转录病毒载体方面获得了丰富的经验，这些载体来源于禽网状内皮组织增生症病毒REV-A和脾坏死病毒SNV，它们在病毒表面展示单链抗体（scA）或其他靶向配体。此外，假型REV衍生的载体与嗜神经性狂犬病病毒株的包膜蛋白，使细胞类型特异性基因传递到神经元在体外和体内。该研究项目的主要目标是进一步开发逆转录病毒载体，使细胞类型特异性基因递送到神经元和脑MVEC中。一系列新的逆转录病毒载体将被开发出来，这些载体将携带可用于艾滋病痴呆基因治疗的治疗基因。载体设计将使得治疗基因能够从细胞类型特异性或诱导型启动子表达。这些载体将在体外和体内进行测试。体外测试包括治疗性基因的效率的长期研究、确定神经元或脑MVEC中正常基因表达的变化的微阵列测定以及体外血脑屏障系统中载体的测试。穆赫塔尔博士在这方面有着丰富的经验。将在小鼠中进行体内实验，以测试长期基因表达，并确定表达治疗基因的动物大脑中是否发生宏观或微观变化。开发神经元和脑MVECs特异性的细胞类型特异性载体不仅对未来可能应用于AIDS痴呆的基因治疗有用，而且对许多其他CNS疾病也有用。

DESCRIPTION (provided by applicant): Human immunodeficiency virus type- I (HIV-1) infection can lead to a series of devastating clinical conditions in the central nervous system (CNS) of certain HIV-1-infected individuals. AIDS dementia complex (ADC) is a collective term to describe AIDS-related cognitive dysfunction, motor difficulties, coordination abnormalities and other neurological signs and symptoms. Blood-brain barrier mainly constituted of micro vascular endothelial cells (MVECs) a protective sheath is a potential obstacle for conventional antiretroviral penetration into the brain. Gene therapy is one the most promising avenues for treatment of neurodegenerative disorders in general and AIDS dementia in particular. Our laboratories in recent past have utilized retroviral vectors for gene delivery in CNS based cells both in vitro and in vivo. Cell-type-specific gene delivery into distinct cells of the central nervous system will be one of the major prerequisites for successful human gene therapy of neurological disorders. In the past decade, Dr. Dornburg's laboratory has gained extensive experience in the construction of cell-type-specific retroviral vectors, derived from the avian reticuloendotheliosis viruses REV-A and spleen necrosis virus, SNV, which display single chain antibodies (scAs) or other targeting ligands on the viral surface. Moreover, pseudotyping REV-derived vectors with the envelope protein of a neurotropic rabies virus strain enabled cell-type-specific gene delivery into neurons in vitro and in vivo. The main goals of this research project are the further development of retroviral vectors, which enable cell-type-specific gene delivery into neurons and brain MVECs. A series of novel retroviral vectors will be developed which transduce therapeutic genes useful for gene therapy of AIDS dementia. The vector design will enable expression of the therapeutic gene from cell-type-specific or inducible promoters. These vectors will be tested in vitro and in vivo. In vitro testing include long-term studies of the efficiency of the therapeutic gene, microarray assays to determine changes of normal gene expression in neurons or brain MVECs, and the testing of the vectors in vitro blood brain barrier systems. Dr. Mukhtar has extensive experience in this area. In vivo experiments will be performed in mice to test long-term gene expression and to determine whether macroscopic or microscopic changes occur in the brains of animals which express the therapeutic genes. The development of cell-type-specific vector specific for neurons and brain MVECs will not only be useful for possible future application of gene therapy of AIDS dementia, but also for numerous other disorders of the CNS.