mRNA Splicing Modulation in Familial Dysautonomia

家族性自主神经功能异常（FD）是一种由剪接引起的遗传性感觉和自主神经病变 IKBKAP基因突变。该突变导致IKBKAP mRNA中外显子20的可变跳跃， 导致IKAP蛋白的组织特异性减少。我们已经表明，剪接是特别差的，在 神经系统，导致IKAP蛋白水平显着降低。尽管FD是隐性的， 患者保留了制造正常mRNA和蛋白质的能力，这一发现提供了一个令人兴奋的，直接 通过剪接修饰开发治疗方法。作为NINDS赞助的 神经生成药物筛选联盟，我们确定激动素，一种植物细胞分裂素，作为一种有效的调节剂， mRNA剪接。该化合物具有显著的功效，并且可以在患者中恢复正常的IKAP蛋白水平。 培养一周内的细胞。最近，我们也证明了激动素可以修饰IKBKAP剪接 在转基因小鼠和人FD携带者和患者中的体内。在过去的三年里，我们 与NINDS蓝图神经治疗网络合作，进行药物化学和优化 激动素，目的是开发一种新的剪接调节剂化合物（SMC）作为FD的治疗。到 迄今为止，我们已经评估了520多种化合物，并确定了200多种化合物， 提高效力和功效。这项资助的目标是详细研究我们的新一类SMC如何改变 mRNA剪接。我们还将对我们的一种SMC进行临床前试验，以首次确定 通过修饰体内剪接增加IKAP蛋白将改善新FD小鼠的疾病表型 模型最后，我们将确定关键疾病相关网络。这将产生未来的潜在生物标志物 临床研究，新的靶向治疗途径，也将阐明调节感觉和 自主神经系统发育尽管FD是一种发育障碍， 一生都在遭受持续剧烈的神经元退化的困扰。我们认为，有效增加 生命早期的IKAP水平可能支持神经元存活，并预防或延迟衰弱性步态、感觉和神经功能障碍。 随着年龄的增长，患者的认知能力下降。成功完成这项赠款的目的将使我们能够 了解我们的SMC的作用机制，确定它们的作用在新动物中是否有效 疾病的模型，并揭示对IKAP蛋白治疗性增加有反应的基因网络。 这些都是我们继续向诊所攀登的基本步骤。

Familial dysautonomia (FD) is a hereditary sensory and autonomic neuropathy that is caused by a splice mutation in the IKBKAP gene. The mutation results in variable skipping of exon 20 in IKBKAP mRNA, which leads to a tissue-specific reduction of IKAP protein. We have shown that splicing is particularly poor in the nervous system, resulting in dramatically reduced levels of IKAP protein. Despite the fact that FD is recessive, patients retain the capacity to make both normal mRNA and protein, a discovery that offered an exciting, direct approach towards the development of therapies via splicing modification. As part of an NINDS sponsored Neurogeneration Drug Screening Consortium, we identified kinetin, a plant cytokinin, as a potent modulator of mRNA splicing. This compound has remarkable efficacy and can restore normal IKAP protein levels in patient cells within one week in culture. Recently, we have also demonstrated that kinetin can modify IKBKAP splicing in vivo in both transgenic mice and in human FD carriers and patients. Over the past three years, we have worked with the NINDS Blueprint Neurotherapeutics Network to perform medicinal chemistry and optimization of kinetin, with the goal of developing a novel splicing modulator compound (SMC) as a therapy for FD. To date, we have evaluated over 520 compounds and have identified more than 200 that have dramatically improved potency and efficacy. The goal of this grant is to study, in detail, how our new class of SMCs modify mRNA splicing. We will also perform a pre-clinical trial of one of our SMCs to determine, for the first time, if increasing IKAP protein by modifying splicing in vivo will improve disease phenotypes in a new FD mouse model. Lastly, we will identify critical disease-relevant networks. This will yield potential biomarkers for future clinical studies, new targetable pathways for therapy, and will also shed light on the regulation of sensory and autonomic nervous system development. Despite the fact that FD is a developmental disorder, patients are plagued by continued, drastic neuronal degeneration throughout life. We believe that effectively increasing IKAP levels early in life may support neuronal survival and prevent or delay the debilitating gait, sensory, and cognitive decline seen in patients as they age. Successful completion of the Aims of this grant will allow us to understand the mechanism of action of our SMCs, determine if their action is efficacious in a novel animal model of the disease, and uncover gene networks that respond to a therapeutic increase in IKAP protein. These are all fundamental steps as we continue the long climb towards the clinic.