Optimization of compounds to improve mRNA splicing in familial dysautonomia

优化化合物以改善家族性自主神经功能障碍中的 mRNA 剪接

• 批准号: 8726499
• 负责人: Susan A Slaugenhaupt
• 金额: $ 19.47万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8726499
• 关键词: Age; Alleles; Biological Assay; Biological Models; Blood - brain barrier anatomy; Brain; Cell Line; Cells; Chemicals; Clinic; Complex; Cytokinins; Data; Development; Disease; Dose; Drug Targeting; Dysautonomias; Exons; Familial Dysautonomia; Family; Firefly Luciferases; Foundations; Funding; Gait; Genes; Goals; Hereditary Disease; Hereditary Sensory Neuropathy; Human; Human Genetics; Impaired cognition; Investments; Joints; Kinetins; Laboratories; Lead; Length; Life; Luciferases; Messenger RNA; Modeling; Modification; Molecular Target; Mus; Mutation; National Institute of Neurological Disorders and Stroke; Nerve Degeneration; Neurons; Neurosciences Research; Oral; Patients; Pattern; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase; Plague; Plants; Preclinical Drug Evaluation; Production; Protein Isoforms; Proteins; RNA Splicing; Reaction; Renilla; Reporter; Resources; Secondary to; Sensory; Structure-Activity Relationship; System; Testing; Time; Tissue Model; Tissue-Specific Splicing; Tissues; Transgenic Mice; United States National Institutes of Health; Work; analog; autonomic neuropathy; cell type; developmental disease; disease-causing mutation; drug development; effective therapy; exon skipping; improved; in vivo; lymphoblast; mRNA Precursor; member; mouse model; nervous system disorder; neuronal survival; novel; prevent; programs; protein expression; public health relevance; response; screening; therapy development

DESCRIPTION (provided by applicant): 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. Analysis of tissues from FD patients shows significantly more exon-skipping in neuronal tissue, and therefore lower IKAP levels. IKAP is a member of the human Elongator complex, which is required for efficient transcriptional elongation of a subset of genes. Despite the fact that FD is recessive, we have shown that patients retain the capacity to make both normal mRNA and protein. This discovery offers an exciting, direct approach towards the development of therapies aimed at increasing levels of cellular IKAP via splicing modification. As part of the NINDS-sponsored Neurodegeneration Drug Screening Consortium, we found that treatment of cultured FD cells with kinetin, a plant cytokinin, enhances exon-20-inclusion and dramatically increases the amount of wild-type IKBKAP mRNA and IKAP protein in FD cells. This compound has remarkable efficacy and can restore normal IKAP protein levels in patient cells within one week in culture. We have demonstrated kinetin's ability to alter IKBKAP splicing using minigene assays in a variety of cell types, as well as in human cells. More recently, we have shown in vivo efficacy in both transgenic mice and in human FD carriers. Despite substantial investment by the Dysautonomia Foundation in developing kinetin as a potential treatment for FD, the road to the clinic has been slow, and we are still working with the originally identified compound. We have recently generated some promising SAR (structure and activity relationship) data showing that the activity of the lead compound kinetin can be improved. It is crucial that chemical optimization be performed in order to improve the potency and activity of kinetin, and time is of the essence. Despite the fact that FD is a developmental disorder, patients are plagued by continued, drastic neuronal degeneration throughout life. Effectively increasing IKAP levels early in life may support neuronal survival and prevent or delay the debilitating gait and sensory and cognitive decline seen in patients as they age. The Blueprint Neurotherapeutics Network offers a unique opportunity for drug development that will provide access to resources that are currently out of reach.

描述（由申请人提供）：家族性自主神经功能障碍（FD）是一种遗传性感觉和自主神经病，由 IKBKAP 基因剪接突变引起。该突变导致 IKBKAP mRNA 中外显子 20 的可变跳跃，从而导致 IKAP 蛋白的组织特异性减少。对 FD 患者组织的分析显示，神经元组织中的外显子跳跃明显增多，因此 IKAP 水平较低。 IKAP 是人类延伸复合体的成员，它是基因子集有效转录延伸所必需的。尽管 FD 是隐性的，但我们已经证明患者保留了产生正常 mRNA 和蛋白质的能力。这一发现为开发旨在通过剪接修饰提高细胞 IKAP 水平的疗法提供了一种令人兴奋的直接方法。作为 NINDS 赞助的神经退行性疾病药物筛选联盟的一部分，我们发现用激动素（一种植物细胞分裂素）处理培养的 FD 细胞可以增强外显子 20 的包含，并显着增加 FD 细胞中野生型 IKBKAP mRNA 和 IKAP 蛋白的数量。该化合物具有显着的功效，可以在培养一周内恢复患者细胞中正常的IKAP蛋白水平。我们通过小基因检测在多种细胞类型以及人类细胞中证明了激动素改变 IKBKAP 剪接的能力。最近，我们在转基因小鼠和人类 FD 携带者中显示了体内功效。尽管自主神经功能障碍基金会投入了大量资金来开发激动素作为 FD 的潜在治疗方法，但通往临床的道路却进展缓慢，我们仍在研究最初确定的化合物。我们最近生成了一些有前景的 SAR（结构和活性关系）数据，表明先导化合物激动素的活性可以得到改善。为了提高激动素的效力和活性，进行化学优化至关重要，而时间至关重要。尽管 FD 是一种发育障碍，但患者一生都受到持续、剧烈的神经元变性的困扰。生命早期有效提高 IKAP 水平可能支持神经元存活和 预防或延缓患者随着年龄增长而出现的步态衰弱以及感觉和认知能力下降。蓝图神经治疗网络为药物开发提供了独特的机会，可以提供目前无法获得的资源。