High throughput screening and in vivo testing of drugs to enhance remyelination

描述(由申请人提供)：多发性硬化症(MS)是一种复杂的、令人衰弱的神经疾病，与显著的发病率和死亡率有关。疾病的病因是未知的环境和遗传因素的结果，而疾病的病理表现为中枢神经系统(CNS)的炎症性损伤，导致身体丧失能力。这种损伤是自身免疫介导的少突胶质细胞破坏导致脱髓鞘的结果。根据疾病的表现，长期脱髓鞘导致轴突损伤发生在局灶性病变部位和/或广泛分布于整个中枢神经系统。MS使用针对免疫系统的疾病改良剂进行治疗，以试图减少复发的频率并延缓疾病的进展。这些药物在改善患者预后方面的成功有限，而且缺乏促进立即重新髓鞘形成的未得到满足的需求。识别诱导常驻少突胶质前体细胞(OPC)介导的重新髓鞘形成的药物将为患者提供阻止疾病进展和改善功能的重要机制。我们建议在多发性硬化症中筛选促进髓鞘再分化的药物。为了实现这一目标，我们制定了一项建议，通过一条经过验证的验证管道，整合了多学科的技能和专业知识，以审问促进髓鞘再分化的药物实体。我们将从在人类临床试验中有使用历史的药物开始，并在我们新的高通量体外系统中评估它们促进啮齿动物OPC成熟和髓鞘基因表达的能力。该系统利用我们的技术，从多能干细胞中产生可扩展的功能性OPC来源。从我们的高通量体外筛选中验证的药物命中将在使用器官型啮齿动物脑片培养的易处理和经济的体外髓鞘形成试验中进行测试。接下来，我们将采用我们最有希望的5种药物(从体外和体外测试确定)，并在动物模型系统中测试它们，这些动物模型系统通常用于询问MS(溶血磷脂介导的脊髓脱髓鞘和实验性自身免疫性脑脊髓炎)的适用性。最热门的药物也将在人类OPC上进行测试。利用我们在临床前多发性硬化症研究和干细胞分化方面的优势，我们的目标是在这个为期两年的项目结束前找到促进髓鞘再生的有效药理制剂。我们准备将从我们的研究中确定的先导药物推进到人体临床试验。

DESCRIPTION (provided by applicant): Multiple sclerosis (MS) is a complex, debilitating neural disorder associated with significant morbidity and mortality. Disease etiology is a result o unknown environmental and genetic factors, while disease pathology presents as inflammatory injury to the central nervous system (CNS) causing physical incapacity. This damage is a result of autoimmune-mediated destruction of oligodendrocytes causing demyelination. Prolonged demyelination leads to axonal damage occurring either in focal lesion sites and/or widespread throughout the CNS, depending on presentation of the disease. MS is treated with disease-modifying agents that target the immune system in an attempt to reduce the frequency of relapses and delay disease progression. These drugs have had limited success in improving patient outcomes and lack the unmet need to promote immediate remyelination. Identifying drugs that induce resident oligodendrocyte progenitor cell (OPC) mediated remyelination would provide patients a vital mechanism to halt disease progression and improve function. We propose to screen drugs that promote remyelination for repurposing in MS. To achieve this goal we have developed a proposal that incorporates the skill and expertise of multiple disciplines through a proven validation pipeline to interrogate drug entities that promote remyelination. We will begin with drugs that have a history of use in human clinical trials and assess their ability o promote rodent OPC maturation and myelin gene expression in our novel high throughput in vitro system. This system takes advantage of our technology to generate scalable sources of functional OPCs from pluripotent stem cells. Validated drug hits from our high throughput in vitro screen will then be tested in a tractable and economical ex vivo myelination assay utilizing organotypic rodent brain slice cultures. Moving forward, we will take our 5 most promising drug hits (as determined from in vitro and ex vivo assays) and test them in animal model systems regularly used to interrogate applicability to MS (lysolecithin-mediated spinal cord demyelination and Experimental Autoimmune Encephalomyelitis). Top drug hits will also be tested on human OPCs. Using our strengths in preclinical MS research and stem cell differentiation, we aim to identify efficacious pharmacological agents that promote remyelination by the end of this two year project. We are prepared to progress lead drugs identified from our studies to human clinical trials.