Gene Therapy of AIDS Dementia

描述（由申请人提供）：人类免疫缺陷病毒I型（HIV-1）感染可导致某些HIV-1感染者中枢神经系统（CNS）出现一系列破坏性的临床症状。艾滋病痴呆复合体（AIDS dementia complex， ADC）是艾滋病相关认知功能障碍、运动困难、协调异常和其他神经系统体征和症状的统称。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进入大脑的潜在障碍。基因治疗是治疗一般神经退行性疾病，特别是艾滋病痴呆症最有前途的途径之一。我们的实验室最近在体外和体内利用逆转录病毒载体在基于中枢神经系统的细胞中进行基因传递。细胞类型特异性基因传递到中枢神经系统的不同细胞将是成功的人类基因治疗神经系统疾病的主要先决条件之一。在过去的十年中，Dornburg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，这些逆转录病毒载体来源于禽网状内皮增生病毒REV-A和脾坏死病毒SNV，它们在病毒表面显示单链抗体（scAs）或其他靶向配体。此外，带有嗜神经狂犬病病毒株包膜蛋白的rev衍生的假分型载体能够在体外和体内将细胞类型特异性基因传递到神经元中。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能够传递到神经元和脑mvec中。一系列新的逆转录病毒载体将被开发出来，这些载体可以转导治疗基因，用于艾滋病性痴呆的基因治疗。载体设计将使治疗基因的表达从细胞类型特异性或诱导启动子。这些载体将在体内和体外进行测试。体外测试包括治疗基因效率的长期研究，微阵列检测以确定神经元或脑MVECs中正常基因表达的变化，以及载体在体外血脑屏障系统中的测试。Mukhtar博士在这一领域拥有丰富的经验。体内实验将在小鼠身上进行，以测试长期基因表达，并确定表达治疗基因的动物大脑是否发生宏观或微观变化。针对神经元和脑mvec的细胞类型特异性载体的开发不仅有助于未来艾滋病痴呆的基因治疗，也有助于许多其他中枢神经系统疾病的基因治疗。

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.