Clinical Investigations On The Etiology And Therapy Of N

N的病因及治疗的临床研究

• 批准号: 7324563
• 负责人: Henry F McFarland
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7324563
• 关键词: Clinical; Investigations; Etiology; Therapy

Gaucher Disease: Our primary interest is to understand the pathogenesis of the neurological involvement in this disease and to test novel therapeutic approaches. We are conducting a randomized controlled trial of OGT 918 (N-butyldeoxynojirimycin, miglustat) as a substrate reduction approach for the treatment of patients with neuronopathic Gaucher disease. We are evaluating primarily the neurological abnormalities, and in particular the slow eye movements of these patients. Thus far, this medication is well tolerated. Results of the 1st year did not show any significant difference between the group on OGT 918 and the group. Final results of this two-year study will be available in FY 2007. We showed for the first time that electron microscopy study of the skin could identify patients with type 2 (acute neuronopathic) Gaucher disease. This finding will enable early diagnosis and institution of appropriate therapy to this devastating form of the disease. Over the past year in collaboration with Dr. Ellen Sidransky we further characterized a large cohort of patients with type 3 (chronic neuronopathic) Gaucher disease. We showed that patients homozygous for the most common neuronopathic mutation have a wide variety of phenotypes. This finding emphasizes the role of genetic modifiers in this disorder. On the other hand, we showed that different mutations could lead to the same phenotype - progressive myoclonic encephalopathy. Fabry Disease: Patients with this second most common lysosomal disorder have a severely painful peripheral neuropathy, and a systemic vasculopathy leading to premature strokes, cardiac disease, and kidney insufficiency. We have been exploring enzyme replacement therapy (ERT) for Fabry disease. Patients on long-term therapy had significant reduction in pain, improvement in sensing cold and warm and in their sweat function. However, ERT did not reduce the incidence of stroke. We therefore performed extensive studies on the pathogenesis of stroke in Fabry disease. Our ongoing hypothesis regarding strokes this disorder is that reactive oxygen species play an important role. We found increased staining for 3-nitrotyrosine in dermal and cerebral blood vessels and elevated nitrotyrosine and myeloperoxidase levels in blood of patients. Myeloperoxidase elevation may be related to our observation of premature atherosclerosis in patients with Fabry disease, a finding confirmed by another group in a mouse model for this disorder. We found that the likelihood of occurrence of cerebral lesions seen on MRI in patients with Fabry disease seems to be modified by alterations of other genetic risk factors for stroke such as interleukin-6, factor V Leiden, protein Z and endothelial nitric oxide synthase. This year we also found that random myeloperoxidase in serum and plasma was significantly elevated in 73 consecutive male patients with Fabry disease. Random serum myeloperoxidase level in men predicted the risk of a Fabry vasculopathy-related event in subsequent years. Long-term enzyme replacement therapy did not reduce myeloperoxidase level or eliminate the risk of vasculopathic events. These observations should contribute to the general understanding of Fabry disease and vasculopathies of the general population. Because of the partial effect of ERT in adult patients, we hypothesized that better results may be obtained when it is initiated in childhood. We therefore completed a 6-month study on ERT in children 7-17 years of age with Fabry disease. We found that ERT was safe, led to improved cardiac, renal and sweat function and lessening of neuropathic pain. This study is continuing in order to determine the long-term effect of ERT in this patient population. Using genomic techniques, we found that neuronal apoptosis inhibitor protein is over-expressed in children with Fabry disease. This finding should help understand the pathogenesis of this disease and serve as a potential marker to monitor response to specific therapies. Using state-of-the-art proteomics techniques, and taking advantage of the specific effects of ERT, we found decreased alpha2-antiplasmin and plasminogen in the blood of patients with Fabry disease. This finding likely indicates a general abnormality in fibrinolysis in this disorder. We completed a genomics study in the mouse model of Fabry disease. We examined the heart, liver and aorta of male Fabry knockout mice 28 weeks of age compared with wild-type mice. Gene expression analyses were performed using microarrays before and after six weekly injections of alpha-galactosidase A. RPGRIP1 was the highest ranked statistically in all 3 organs and its splice variants responded in a unique way to alpha-galactosidase A. ERT tended to not only normalize gene expression, but also specifically modified gene expression in each tissue. We then looked at the data using gene expression correlation graphs as well as structural equation modeling and path analysis. This method allowed us to generate statistical hypotheses regarding mechanisms of disease that will be tested in the laboratory in the future. We initiated a program of active-site chaperone therapy in patients with Fabry disease who have with enhanceable alpha-galactosidase A activity. Work in our laboratory confirmed marked in vitro enhanceability of the deficient enzyme in peripheral blood white cells of some patients by the chemical chaperone. A treatment trial is ongoing in patients with Fabry disease and initial results are promising. The extensive delivery throughout the body of chemical chaperones should have a significant advantage over ERT where the infused enzyme enters into vascular endothelial cells for the most part. This therapeutic approach should be applicable to other disorders where the mutated enzyme is enhanceable. Mucolipidosis IV: We continued our investigation of 35 patients with this autosomal recessive neurogenetic disorder with particular emphasis on their progressive retinal degeneration. The latter is thought to be caused by death of neurons in the retina. Using electroretinography and visual evoked potential we quantified the decline of retinal function in MLIV. This study will be useful to identify the optimal stage for therapeutic intervention to prevent progression of the retinal dystrophy in MLIV. The neurological aspect of this disorder is developmental in nature with little decline over the years. It is therefore evident that mucolipidosis IV is both a developmental and a degenerative disorder. The presentation as a cerebral palsy-like encephalopathy often delays the correct diagnosis of this condition. Leukodystrophies: We continued studying patients with leukodystrophies including the fatal leukodystrophy CACH (childhood ataxia with CNS hypomyelination), a leukodystrophy that was first identified by DMNB in 1992. It is caused by a deficiency of eukaryotic initiation factor 2B (eIF2B). The clinical decline is often initiated or worsens after stress such as a mild head trauma or a febrile illness. eIF2B is a protein complex that is essential for the regulation of protein synthesis, particularly in response to stress. Over the past year we continued collaboration on a finding that patients with eIF2B mutations have marked decrease of asialo-transferrin in the cerebrospinal fluid. This finding was confirmed in a blinded analysis and was found to have 100% and 98% specificity. The reduction of asialo-transferrin likely reflects disturbance in the function of oligodendrocytes and astrocytes and will help with diagnosis and disease management. We also identified a novel leukodystrophy syndrome consisting of hypomyelination, hypogonadotropic hypogonadism and hypodontia (termed 4H syndrome). Electron microscopy and myelin protein immunohistochemistry of sural nerve showed unique findings.

Gaucher病：我们的主要兴趣是了解该疾病中神经学参与的发病机理并测试新颖的治疗方法。我们正在进行OGT 918（N-丁基氧基诺二霉素，Miglustat）的随机对照试验，作为一种用于治疗神经疾病Gaucher病患者的底物还原方法。我们主要评估神经系统异常，尤其是这些患者的眼睛运动缓慢。到目前为止，这种药物的耐受性良好。第一年的结果没有显示OGT 918和该组的组之间的任何显着差异。这项为期两年的研究的最终结果将在2007财年提供。我们首次表明，对皮肤的电子显微镜研究可以鉴定患有2型（急性神经性）Gaucher病的患者。这一发现将使早期诊断和对这种毁灭性疾病形式进行适当的治疗。在过去的一年中，与艾伦·西德兰斯基（Ellen Sidransky）博士合作，我们进一步表征了大量3型患者（慢性神经性疾病）Gaucher病。我们表明，最常见的神经性突变纯合患者具有多种表型。这一发现强调了遗传修饰剂在该疾病中的作用。另一方面，我们表明不同的突变可能导致相同的表型 - 进行性肌阵挛性脑病。 Fabry病：第二大常见溶酶体疾病患者患有严重疼痛的周围神经病，并且系统性血管病导致过早中风，心脏病和肾脏不足。我们一直在探索用于法布里病的酶替代疗法（ERT）。长期治疗的患者的疼痛显着减轻，感测冷，温暖和汗水功能的改善。但是，ERT并没有降低中风的发生率。因此，我们对Fabry病中卒中的发病机理进行了广泛的研究。我们正在进行的关于这种疾病的假设是活性氧起着重要作用。我们发现，在皮肤和脑血管中3-硝基酪氨酸的染色增加，硝基酪氨酸和髓质过氧化物酶水平升高。骨髓过氧化物酶升高可能与我们观察到Fabry疾病患者过早动脉粥样硬化有关，这一发现由小鼠模型中的另一组证实了该疾病。我们发现，Fabry病患者在MRI上出现大脑病变的可能性似乎是通过其他中风的其他遗传危险因素（例如白介素-6，因子V Leiden，蛋白质Z和蛋白质Z和内皮氮氧化物合酶）的改变。今年，我们还发现，连续73例Fabry疾病患者，血清和血浆中的随机脊髓过氧化物酶显着升高。男性的随机血清髓过氧化物酶水平预测了随后几年发生Fabry Vasculopathy相关事件的风险。长期酶​​替代疗法并未降低髓过氧化物酶水平或消除血管性事件的风险。这些观察结果应有助于对Fabry病和普通人群的血管病的一般理解。由于ERT对成年患者的部分影响，我们假设在儿童期开始时可以获得更好的结果。因此，我们完成了7-17岁的Fabry病儿童的6个月研究。我们发现ERT是安全的，导致心脏，肾脏和汗水功能改善，并减轻神经性疼痛的减轻。为了确定ERT在该患者人群中的长期影响，这项研究正在继续。使用基因组技术，我们发现神经元凋亡抑制剂蛋白在Fabry疾病的儿童中过表达。这一发现应有助于了解该疾病的发病机理，并成为监测对特定疗法反应的潜在标记。使用最先进的蛋白质组学技术并利用ERT的特定作用，我们发现Fabry病患者的血液中α2-抗血解剂和纤溶酶原降低。这一发现可能表明该疾病中纤维蛋白溶解的普遍异常。我们在Fabry病小鼠模型中完成了一项基因组学研究。与野生型小鼠相比，我们检查了28周雄性Fabry敲除小鼠的心脏，肝脏和主动脉。在每周六次注射α-半乳糖苷酶A. rpgrip1之前和之后，使用微阵列进行了基因表达分析。RPGRIP1在所有3个器官中统计上排名最高，其剪接变体以一种独特的方式响应α-甲乳酸化酶A. erpha-galactosidase A. ert eRT倾向于不仅倾向于在每个级别的表达中均衡，但在每个组织中都归一化，但在每个组织中都有归一化。然后，我们使用基因表达相关图以及结构方程建模和路径分析查看了数据。这种方法使我们能够产生有关将来将在实验室中测试的疾病机制的统计假设。我们在患有可增强的α-半乳糖苷酶A活性的Fabry疾病患者的患者中启动了一项活动部位伴侣治疗程序。在我们的实验室中的工作证实，通过化学伴侣，某些患者的外周血白细胞中缺乏酶的体外增强了体​​外增强性。 Fabry病患者正在进行一项治疗试验，最初的结果有望。整个化学伴侣体内的广泛递送应该比ERT具有显着优势，在该ERT中，注入的酶在大多数情况下进入血管内皮细胞。这种治疗方法应适用于突变酶可增强的其他疾病。 粘脂脂异常IV：我们继续研究35例这种常染色体隐性神经遗传疾病，特别强调其进行性视网膜变性。后者被认为是由视网膜中神经元死亡引起的。使用电子模拟和视觉诱发电位，我们量化了MLIV中视网膜功能的下降。这项研究将有助于确定治疗干预的最佳阶段，以防止MLIV中视网膜营养不良的进展。多年来，这种疾病的神经方面本质上是发育性的，几乎没有下降。因此，很明显，粘膜脂蛋白IV既是发育性疾病，也是退化性疾病。作为脑瘫样性脑病的表现通常会延迟对这种情况的正确诊断。 白细胞营养不良：我们继续研究白细胞营养不良的患者，包括致命的白细胞营养哲学CACH（儿童性共济失调，具有CNS降低率低），这是一种白细胞营养不良，DMNB于1992年首次鉴定出来。它是由真核启动因子2B（EIF2B）的缺乏引起的。在压力（例如轻度头部外伤或高热疾病）后，临床下降通常会引发或恶化。 EIF2B是一种蛋白质复合物，对于调节蛋白质合成至关重要，尤其是在响应胁迫方面。在过去的一年中，我们继续合作，发现EIF2B突变患者在脑脊液中标志着Asialo-Transferrin明显减少。这一发现在盲目的分析中得到了证实，发现具有100％和98％的特异性。 ASIALO转移蛋白的减少可能反映出少突胶质细胞和星形胶质细胞功能的干扰，并将有助于诊断和疾病管理。我们还确定了一种新型的白细胞营养不良综合征，该综合征包括低monse骨，性腺功能减退和低音症（称为4H综合征）。电子显微镜和髓磷脂蛋白免疫组织化学的表面神经显示出独特的发现。