Macrophage-mediated gene delivery of neurotrophic factors for Parkinson's disease

巨噬细胞介导的帕金森病神经营养因子基因传递

• 批准号: 8244211
• 负责人: SENLIN LI
• 金额: --
• 依托单位: SOUTH TEXAS VETERANS HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8244211
• 关键词: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Acute; Address; Adrenoleukodystrophy; Adult; Adverse effects; Affect; Age; Animal Model; Antibodies; Area; Biological Availability; Blood; Blood - brain barrier anatomy; Bone Marrow; Bone Marrow Cell Transplantation; Bone Marrow Cells; Brain; Bypass; Cannulas; Catheters; Cell Nucleus; Cells; Characteristics; Chronic; Clinical; Clinical Research; Clinical Trials; Corpus striatum structure; Defect; Development; Disease; Disorder by Site; Dopamine; Doxycycline; Elderly; Emotional; Engineering; Fiber; Future; Gene Delivery; Gene Expression; Genetic; Genetic Models; Health Care Costs; Hematopoietic stem cells; Homing; Human; Infusion procedures; Injury; Isogenic transplantation; Lead; Lentivirus Vector; Literature; Location; Long-Term Effects; Mediating; Membrane Transport Proteins; Methods; Microglia; Modeling; Monitor; Monkeys; Mus; Mutagenesis; Nerve Degeneration; Neurodegenerative Disorders; Neurologic; Neurons; Neurotoxins; Operative Surgical Procedures; Parkinson Disease; Pathology; Pathway interactions; Patients; Penetration; Population; Probenecid; Problem Solving; Property; Proteins; Recruitment Activity; Relative (related person); Safety; Simulate; Site; Solid; Staging; Substantia nigra structure; Switch Genes; Symptoms; System; Technology; Testing; Tetanus Helper Peptide; Tetracyclines; Therapeutic; Time; Toxin; Transgenic Organisms; Translating; Transplantation; Treatment Efficacy; Viral Vector; Withdrawal; axon regeneration; base; brain tissue; clinical application; clinically relevant; design; dopaminergic neuron; gene therapy; glial cell-line derived neurotrophic factor; improved; in vivo; knockout gene; macrophage; mitopark mouse; motor deficit; mouse model; neuroprotection; neurorestoration; neurotrophic factor; novel; novel strategies; pre-clinical; promoter; putamen; research clinical testing; success; targeted delivery; transgene expression; translational study

DESCRIPTION (provided by applicant): The development of a neuroprotective/restorative therapy for Parkinson's disease (PD) would be a major therapeutic advance. Glial cell line-derived neurotrophic factor (GDNF) has been the most potent trophic molecule tested so far and in numerous studies has clearly demonstrated a capacity to protect and restore neurons affected in PD. However, clinical trials of GDNF in PD patients have given mixed results. Effective targeted delivery to specific brain sites of disease is believed to be the key impediment to consistent success. GDNF requires focal delivery as it does not cross the blood- brain barrier. The precise location of the cannula for delivery is critical and it is challenging to achieve therapeutic levels of GDNF for all or most degenerating neurons due to the relatively large target area in human brain and the poor brain tissue penetration of this molecule. Additionally, current GDNF therapy entails important safety concerns. We recently developed a novel approach - macrophage- mediated GDNF delivery - that seems capable of resolving these problems. This unique approach takes advantage of the well-known macrophage property of homing to degenerating sites in proximity to damaged neurons, capitalizes on our powerful macrophage-specific synthetic promoters (MSP), and implements recent advances in hematopoietic stem cell gene therapy. We hypothesize that highly effective CNS delivery of GDNF can be achieved through its expression in macrophages / microglia by ex vivo transduction of HSC-containing bone marrow cells with lentiviral vectors carrying a cassette expressing GDNF driven by our MSP, followed by syngeneic transplantation of these transduced bone marrow cells, and this will greatly ameliorate the pathological changes and neurological defects of PD. Using a sub-acute MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) mouse model of PD in a neuroprotection paradigm, we demonstrated that genetically engineered bone marrow cell-derived macrophages accumulate in diseased sites and macrophage-mediated GDNF delivery dramatically reduces degeneration of dopaminergic neurons of the substantia nigra, as well as their fibers in the striatum, and induces axon regeneration, without any apparent adverse effects. In order to move this novel concept and unique approach eventually into clinical application, here we propose to explore it in depth in both neurotoxin and genetic mouse models featuring the chronic and progressive changes characteristic of PD. In the chronic toxin model, we will utilize a tetracycline-regulatable (Tet-on) system and address efficacy of our approach by doxycycline-induced switching on of GDNF expression and delivery in a neurorestorative or neuroprotective / restorative paradigm, simulating treatment of PD. In the MitoPark conditional gene knockout model, we will perform transplantation of the transduced bone marrow cells at ages of the mice when they reach the neurodegenerative stages equivalent to preclinical, clinical, and advanced PD. We will also address the safety issue by monitoring for adverse effects, particularly those described in the literature, after short- or long-term GDNF expression, and examining the capability of tapering or shutting off GDNF expression when necessary through withdrawal of doxycycline. This translational study will establish a solid base for future clinical investigation of the potential benefits to patients of this novel neuroprotective therapy for PD.

描述（由申请人提供）： 开发针对帕金森氏病（PD）的神经保护性/恢复性疗法将是一个重大的治疗疗法。到目前为止，神经胶质细胞系衍生的神经营养因子（GDNF）是测试的最有效的营养分子，在许多研究中，清楚地表明了保护和恢复PD中受影响的神经元的能力。但是，PD患者中GDNF的临床试验给出了不同的结果。据信，有效地靶向疾病的特定大脑部位是始终取得成功的关键障碍。 GDNF需要局灶性递送，因为它不会越过血脑屏障。插管的确切位置至关重要，由于人脑中的靶标相对较大，该分子的脑组织渗透较差，因此为所有或大多数退化神经元获得治疗水平的GDNF水平至关重要。此外，当前的GDNF疗法还包括重要的安全问题。我们最近开发了一种新型的方法 - 巨噬细胞介导的GDNF递送 - 似乎能够解决这些问题。这种独特的方法利用了与受损神经元的邻近归因地点的众所周知的巨噬细胞特性，利用了我们强大的巨噬细胞特异性合成启动子（MSP），并实现了血肿干细胞基因治疗的最新进展。 We hypothesize that highly effective CNS delivery of GDNF can be achieved through its expression in macrophages / microglia by ex vivo transduction of HSC-containing bone marrow cells with lentiviral vectors carrying a cassette expressing GDNF driven by our MSP, followed by syngeneic transplantation of these transduced bone marrow cells, and this will greatly ameliorate the pathological changes and PD的神经缺陷。 Using a sub-acute MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) mouse model of PD in a neuroprotection paradigm, we demonstrated that genetically engineered bone marrow cell-derived macrophages accumulate in diseased sites and macrophage-mediated GDNF delivery dramatically reduces degeneration of dopaminergic neurons of the黑质及其在纹状体中的纤维及其纤维，并诱导轴突再生，没有任何明显的不利影响。为了将这种新颖的概念和独特的方法最终转移到临床应用中，我们建议在神经毒素和遗传小鼠模型中深入探索它，具有PD的慢性和渐进变化特征。在慢性毒素模型中，我们将利用四环素调节（TET-ON）系统，并通过强力霉素引起的神经训练或神经保护性 /神经保护性 /恢复性态度的GDNF表达和传递来解决我们的方法的疗效，模拟PD的处理。在Mitopark条件基因基因敲除模型中，当它们达到临床前，临床和晚期PD的神经退行性阶段时，我们将对小鼠年龄的转导骨髓细胞进行移植。我们还将通过监测短期或长期GDNF表达后的不良影响，尤其是文献中描述的不良影响，并检查逐渐变细或在必要时通过撤回强力霉素来检查GDNF表达的能力，来解决安全问题。这项翻译研究将建立一个坚实的基础，以对这种新型神经保护疗法对PD的潜在益处进行临床研究。