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

基本信息

批准号：
8034706
负责人：
YUANAN LU
金额：
$ 33.83万
依托单位：
UNIVERSITY OF HAWAII AT MANOA
依托单位国家：
美国
项目类别：
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/8034706
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-1 Health Homing 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 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 Virus base combat gene therapy genetically modified cells insight interest macrophage migration monocyte mouse model nervous system disorder neurotoxic novel strategies programs receptor expression research study therapeutic gene uptake vector

项目摘要

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）：人类免疫缺陷病毒1型（HIV）感染了脑巨噬细胞和小胶质细胞，并引起HIV-与HIV相关的痴呆症（HAT），这是中枢神经系统（CNS）的原发性疾病，影响了约20％的HIV感染者。许多抗逆转录病毒药物越过血脑屏障（BBB）的效率较差，对当前的治疗方法受到了阻碍。因此，需要新的疗法。众所周知，循环血单核细胞衍生的巨噬细胞（MDM）在正常情况下和某些情况下都迁移到BBB上，并进入中枢神经系统；随后，有些人将其成熟到长寿命的组织脑巨噬细胞和小胶质细胞中。这项研究的假设是，有可能利用血液MDM的自然归巢/迁移特性，以便以非侵入性和非手术方式将神经保护因子传递到中枢神经系统中。我们实验室和其他人的先前研究表明，使用有缺陷的慢病毒载体（DLV）可以在体外对MDM（人和小鼠）进行基因修饰，而不会对其生物学特性产生不利影响。此外，我们已经表明，通过DLV遗传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，并且可以通过瞬时BBB来增强CNS摄取这些细胞的效率。我们最近表明，来自DLV转导的MDM的分泌的抗HIV-TAT单链抗体（SCFV）和可溶性肿瘤坏死因子受体（STNFR）可以有效地阻止HIV-1感染细胞或相应重组蛋白的调节培养基的神经毒性作用。在这个项目中，我们将建立一种高效率的基因转移方法（> 50％），用编码可分泌的SCFV和STNFR的向量对主要小鼠MDM的稳定转导。然后，我们将进行研究，以确定这些遗传修饰的细胞是否可以通过暂时破坏BBB来增强小鼠脑，并在外周外插入遗传学改性的MDM后评估CNS基因表达的时间过程和稳定性。最后，我们将确定稳定表达TNFR或抗HIV-TAT SCFV的原代小鼠MDM是否可以改善神经辅助的两个经过良好研究的鼠模型系统中的疾病进展。预计这项研究的成功完成将为对抗神经助剂和其他神经系统疾病的新方法提供重大见解。公共卫生相关性：预计这项研究的成功完成将为对抗神经助剂和其他神经系统疾病的新方法提供重大见解。