Modeling monocyte and macrophage based gene therapy for neuroAIDS

基于单核细胞和巨噬细胞的神经艾滋病基因治疗模型

• 批准号: 7685558
• 负责人: YUANAN LU
• 金额: $ 34.18万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7685558
• 关键词: AIDS Dementia Complex; AIDS neuropathy; Acquired Immunodeficiency Syndrome; Affect; Animal Model; Animals; Anti-Retroviral Agents; Antibodies; Biological; Biological Models; Blood - brain barrier anatomy; Bone Marrow; Brain; Cells; Central Nervous System Diseases; Conditioned Culture Media; Dementia; Disease; Disease Progression; Evaluation; Gene Expression; Gene Transfer; Genes; HIV; HIV-1; Human; In Vitro; Individual; Inflammatory Response; Infusion procedures; Intravenous infusion procedures; Laboratories; Lentivirus Vector; Life; Macrophage Activation; Measures; Mediating; Methods; Microglia; Modeling; Mononuclear; Mus; Neuraxis; Neurocognitive; Neurons; Neurotoxins; Operative Surgical Procedures; Peripheral; Phagocytes; Pharmaceutical Preparations; Physiological; Preparation; Principal Investigator; Production; Property; Recombinant Proteins; Research; Satellite Viruses; Series; System; TNFR-Fc fusion protein; Testing; Time; Tissues; Toxic effect; Transduction Gene; Transgenes; Transgenic Organisms; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Virus; base; combat; gene therapy; genetically modified cells; insight; interest; macrophage; migration; monocyte; mouse model; nervous system disorder; neurotoxic; novel strategies; programs; public health relevance; receptor; receptor expression; research study; therapeutic gene; uptake; vector

DESCRIPTION (provided by applicant): Human immunodeficiency virus type 1 (HIV) infects brain macrophages and microglia and causes HIV- associated dementia (HAD), a primary disorder of the central nervous system (CNS) that affects about 20% of HIV-infected individuals. Current treatment of HAD is hampered by the poor efficiency of many antiretroviral drugs to cross the blood-brain barrier (BBB). Hence, new therapies for HAD are needed. Circulating blood monocyte-derived macrophages (MDM) are known to migrate across the BBB and to enter the CNS under both normal and certain circumstances; some subsequently maturing into long-lived tissue-resident brain macrophages and microglia. The hypothesis of this research is that it may be possible to exploit the natural homing/migratory properties of blood MDM in order to deliver neuroprotective factors into the CNS, in a non- invasive and non-surgical manner. Previous studies from our laboratory and others, have shown that MDM (human and mouse) can be genetically modified in vitro using defective lentiviral vectors (DLV) without adversely affecting their biological properties. Furthermore, we have shown that primary mouse blood monocytes and bone marrow-derived macrophages genetically modified by DLV can enter the brain, and that the efficiency of CNS uptake of these cells can be enhanced through transient disruption of the BBB. We have recently shown that secreted anti-HIV-Tat single chain antibodies (scFv) and soluble tumor necrosis factor receptor (sTNFR) from DLV-transduced MDM can effectively block the neurotoxic effects of conditioned medium from HIV-1 infected cells or respective recombinant proteins. In this project, we will establish a gene transfer method for high efficiency (>50%), stable transduction of primary mouse MDM with vectors that encode secretable scFv and sTNFR. We will then conduct studies to determine whether uptake of these genetically modified cells into the mouse brain can be enhanced using temporary disruption of the BBB, and we will evaluate the time course and stability of intra-CNS gene expression following peripheral infusion of the genetically-modified MDM. Finally, we will determine whether primary mouse MDM that stably express TNFR or anti-HIV-Tat scFv can ameliorate disease progression in two well-studied murine model systems for neuroAIDS. Successful completion of this study is expected to provide significant insight into new approaches for combating neuroAIDS and other neurologic disorders. PUBLIC HEALTH RELEVANCE: Successful completion of this study is expected to provide significant insight into new approaches for combating neuroAIDS and other neurologic disorders.

描述（由申请人提供）：1 型人类免疫缺陷病毒 (HIV) 感染大脑巨噬细胞和小胶质细胞并导致 HIV 相关痴呆 (HAD)，这是一种中枢神经系统 (CNS) 的原发性疾病，影响约 20% 的 HIV 感染者个人。目前 HAD 的治疗因许多抗逆转录病毒药物穿过血脑屏障 (BBB) 的效率较低而受到阻碍。因此，需要针对 HAD 的新疗法。众所周知，在正常和某些情况下，循环血液中的单核细胞衍生巨噬细胞 (MDM) 会迁移穿过 BBB 并进入 CNS；一些随后成熟为长寿的组织驻留脑巨噬细胞和小胶质细胞。本研究的假设是，有可能利用血液 MDM 的自然归巢/迁移特性，以非侵入性和非手术方式将神经保护因子输送到 CNS 中。我们实验室和其他实验室之前的研究表明，可以使用有缺陷的慢病毒载体 (DLV) 在体外对 MDM（人类和小鼠）进行基因改造，而不会对其生物学特性产生不利影响。此外，我们还发现，经过 DLV 基因修饰的原代小鼠血液单核细胞和骨髓来源的巨噬细胞可以进入大脑，并且通过短暂破坏 BBB 可以提高中枢神经系统摄取这些细胞的效率。我们最近发现，DLV转导的MDM分泌的抗HIV-Tat单链抗体（scFv）和可溶性肿瘤坏死因子受体（sTNFR）可以有效阻断来自HIV-1感染细胞的条件培养基或各自重组蛋白的神经毒性作用。在该项目中，我们将建立一种基因转移方法，使用编码可分泌型 scFv 和 sTNFR 的载体，实现原代小鼠 MDM 的高效（>50%）稳定转导。然后，我们将进行研究，以确定是否可以通过暂时破坏血脑屏障来增强这些转基因细胞对小鼠大脑的吸收，并且我们将评估外周输注基因修饰细胞后中枢神经系统内基因表达的时间过程和稳定性。修改后的 MDM。最后，我们将确定稳定表达 TNFR 或抗 HIV-Tat scFv 的原代小鼠 MDM 是否可以在两个经过充分研究的神经艾滋病小鼠模型系统中改善疾病进展。这项研究的成功完成预计将为对抗神经艾滋病和其他神经系统疾病的新方法提供重要的见解。公共卫生相关性：这项研究的成功完成预计将为对抗神经艾滋病和其他神经系统疾病的新方法提供重要的见解。