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（MS）是一种复杂的、使人衰弱的神经疾病，与显著的发病率和死亡率相关。疾病病因是未知的环境和遗传因素的结果，而疾病病理表现为中枢神经系统（CNS）的炎性损伤，导致身体机能丧失。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CNS。MS用靶向免疫系统的疾病修饰剂治疗，试图降低复发频率并延迟疾病进展。这些药物在改善患者预后方面的成功有限，并且缺乏促进立即髓鞘再生的未满足需求。鉴定诱导常驻少突胶质祖细胞（OPC）介导的髓鞘再生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。我们建议筛选促进髓鞘再生的药物，以在MS中重新利用。为了实现这一目标，我们制定了一项建议，通过经过验证的验证管道，将多个学科的技能和专业知识结合起来，以询问促进髓鞘再生的药物实体。我们将开始在人类临床试验中使用的药物，并在我们的新型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产生功能性OPC的可扩展来源。来自我们的高通量体外筛选的经验证的药物命中物然后将在利用器官型啮齿动物脑切片培养物的易处理且经济的离体髓鞘形成测定中进行测试。展望未来，我们将采取我们的5个最有前途的药物命中（从体外和离体测定确定），并在动物模型系统中测试它们，这些系统经常用于询问MS（溶血卵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。热门药物也将在人类OPCs上进行测试。利用我们在临床前MS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确定有效的促进髓鞘再生的药理学药物。我们准备将从我们的研究中确定的先导药物进展到人体临床试验。

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.