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感染者的中枢神经系统(CNS)出现一系列破坏性的临床症状。艾滋病痴呆复合体(ADC)是描述艾滋病相关认知功能障碍、运动障碍、协调异常等神经体征和症状的统称。血脑屏障主要由微血管内皮细胞(MVECs)组成，是常规抗逆转录病毒进入脑内的潜在障碍。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方法之一。我们的实验室最近利用逆转录病毒载体在体外和体内将基因输送到基于CNS的细胞中。将特定细胞类型的基因导入中枢神经系统的不同细胞将是成功的人类神经疾病基因治疗的主要先决条件之一。在过去的十年中，多恩伯格博士的实验室在构建特定细胞类型的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组织增生症病毒REV-A和脾坏死病毒SNV，它们在病毒表面显示单链抗体(SCA)或其他靶向配体。此外，带有嗜神经性狂犬病病毒株包膜蛋白的伪分型REV衍生载体能够在体外和体内将特定细胞类型的基因输送到神经元中。该研究项目的主要目标是进一步开发逆转录病毒载体，使特定细胞类型的基因能够传递到神经元和脑微血管内皮细胞。将开发一系列新型逆转录病毒载体，转导可用于艾滋病痴呆基因治疗的治疗基因。载体设计将使治疗性基因能够从细胞类型特定的或可诱导的启动子中表达。这些载体将在体外和体内进行测试。体外测试包括治疗基因有效性的长期研究，确定神经元或脑微血管内皮细胞正常基因表达变化的微阵列分析，以及体外血脑屏障系统中载体的测试。穆赫塔尔博士在这一领域拥有丰富的经验。将在小鼠身上进行活体实验，以测试长期基因表达，并确定表达治疗基因的动物的大脑是否发生宏观或微观变化。针对神经元和脑微血管内皮细胞的细胞类型特异性载体的开发，不仅为未来艾滋病痴呆的基因治疗提供了可能，也为其他多种中枢神经系统疾病的治疗提供了可能。

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.