GSK3 beta study in patients with Myotonic Dystrophy 1

强直性肌营养不良患者的 GSK3 beta 研究 1

• 批准号: 10593112
• 负责人: LUBOV T TIMCHENKO
• 金额: $ 20.2万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-15 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10593112
• 关键词: 19q; 3' Untranslated Regions; Address; Adult; Affect; Antisense Oligonucleotides; Biochemical; Biopsy; Blood; Blood specimen; CDK4 gene; CUG repeat; Childhood; Chromosomes; Clinical Research; Clinical Trials; Complex; Development; Disease; Family; Fibroblasts; Genes; Goals; Growth; Human; Infant; Knowledge; Left; Length; Link; Mediating; Messenger RNA; Modification; Molecular; Molecular Analysis; Monitor; Multicenter Studies; Mus; Muscle; Muscle Cells; Muscle Fibers; Muscle function; Muscular Atrophy; Myoblasts; Myofibroblast; Myopathy; Myotonia; Myotonic Dystrophy; Myotonic dystrophy type 1; Neuromuscular Diseases; Newborn Infant; Pathologic; Pathology; Pathway interactions; Patients; Penetrance; Phase; Phase I/II Clinical Trial; Phenotype; Phosphotransferases; Pilot Projects; Publishing; RNA Processing; RNA Splicing; RNA-Binding Proteins; Severities; Severity of illness; Signal Transduction; Skeletal Muscle; Skin; Solid; Testing; Therapeutic; Time; Treatment outcome; anxiety reduction; biopharmaceutical industry; clinical development; cohort; cyclin D3; glycogen synthase kinase 3 beta; improved; in vivo; inhibitor; mouse model; mutant; novel therapeutic intervention; postnatal; prevent; small molecule; therapeutic target

Myotonic Dystrophy type 1 (DM1) is a complex neuro-muscular disease, caused by an expansion of CTG repeats in the 3’ UTR of DMPK gene on the chromosome 19q. The longest CTG expansions affect infants causing congenital DM1 (CDM1). CTG repeats cause DM1 pathology through alterations of several RNA- binding proteins including CUGBP1 and MBNL1. The phase I/II clinical trial, testing the degradation of the mutant DMPK mRNA by antisense oligonucleotide (AON) has been recently completed. Unfortunately, it was found that further modifications of AONs are required to improve their penetrance to skeletal muscle. Therefore, there is an urgent need for development of alternative therapeutic approaches based on small molecules. My lab is working with the development of a novel therapeutic approach for the treatment of CDM1 and DM1 which is based on the correction of RNA processing (including splicing) that is mis-regulated by CUG n-GSK3β-CUGBP1 axis. We found that the activity of CUGBP1 is regulated by GSK3β-cyclin D3-CDK4 pathway and that the mutant CUG repeats alter CUGBP1 activity by the pathological increase of GSK3β kinase. The inhibitors of GSK3 significantly reduce DM1 muscle pathology in a mouse model of DM1 (HSA mice) and improve growth and neuromotor activity in CDM1 mouse model (DMSXL mice). These basic findings created a strong background for the first, the GSK3-based clinical study for adult patients with CDM1. The Phase IIa of this trial with the inhibitor of GSK3 tideglusib (TG) was recently completed by a biopharmaceutical company AMO Pharma with encouraging results. Although the trial is in transition to the Phase II/III in pediatric CDM1, much more mechanistic studies, including molecular analysis of the GSK3β-CUGBP1 pathway in human patients’ materials, are needed to further support the clinical trial using TG. My application will examine the GSK3β-CUGBP1 pathway in muscle biopsies, myofibrobasts and blood samples from DM1 and CDM1 patients with variable disease severity (Aim 1). We will also study if GSK3β was corrected in CDM1 patients treated with TG. We will determine if the treatments with TG recover MBNL1 activity correcting global splicing abnormalities in DM1 (Aim 2). All pathways affected by TG treatments will be defined. One more critical issue will be addressed in my application. We recently found that the TG-treatments of DM1 myoblasts and fibroblasts derived from patients with 200-400 CTG repeats reduce the levels of the mutant DMPK mRNA. We propose to examine if TG improves degradation of the mutant DMPK regardless of the length of CTG expansions (Aim 3). If TG improves the degradation of the mutant DMPK mRNA in DM1 and CDM1 myoblasts and fibroblasts with long expansions, the inhibitors of GSK3 will represent small molecules targeting the prime cause of CDM1 and DM1. The obtained knowledge will be critical for the further development of clinical trials of CDM1 and DM1 that are based on the correction of GSK3β.

强直性肌营养不良1型（DM 1）是一种复杂的神经肌肉疾病，由CTG扩张引起 在染色体19 q上DMPK基因的3' UTR中重复。最长的CTG扩张影响婴儿 导致先天性DM 1（CDM 1）。CTG重复序列通过几种RNA的改变引起DM 1病理学改变， 结合蛋白包括CUGBP 1和MBNL 1。I/II期临床试验，测试 通过反义寡核苷酸（AON）突变DMPK mRNA最近已经完成。不幸的是， 发现AON需要进一步修饰以提高其对骨骼肌的亲和力。 因此，迫切需要开发基于小分子的替代治疗方法。 分子。我的实验室正在开发一种新的治疗方法来治疗CDM 1 和DM 1，其基于由CUG错误调节的RNA加工（包括剪接）的校正 n-GSK 3 β-CUGBP 1轴。我们发现CUGBP 1的活性受GSK 3 β-cyclin D3-CDK 4的调节 突变的CUG重复序列通过病理性的GSK 3 β增加而改变CUGBP 1的活性 激酶。在DM 1（HSA）小鼠模型中，GSK 3抑制剂显著减轻DM 1肌肉病理 小鼠）中的生长和神经运动活性，并改善CDM 1小鼠模型（DMSXL小鼠）中的生长和神经运动活性。这些基本发现 为第一项针对成人CDM 1患者的基于GSK 3的临床研究创造了强有力的背景。的 最近，一家生物制药公司完成了这项使用GSK 3抑制剂tideglusib（TG）的IIa期试验。 AMO Pharma取得了令人鼓舞的成绩。尽管该试验正在过渡到儿科II/III期 CDM 1，更多的机制研究，包括GSK 3 β-CUGBP 1途径的分子分析， 人类患者的材料，需要进一步支持使用TG的临床试验。我的申请将审查 来自DM 1和CDM 1的肌肉活检、肌成纤维细胞和血液样品中的GSK 3 β-CUGBP 1通路 不同疾病严重程度的患者（目标1）。我们还将研究在CDM 1患者中是否纠正了GSK 3 β 用TG治疗。我们将确定用TG治疗是否恢复MBNL 1活性，纠正整体剪接 DM 1异常（目标2）。将定义受TG治疗影响的所有途径。还有一个关键问题 会在我的申请中提及我们最近发现，TG处理的DM 1成肌细胞和 来自具有200-400个CTG重复的患者的成纤维细胞降低了突变体DMPK mRNA的水平。我们 建议检查TG是否改善突变体DMPK的降解，而不管CTG的长度如何 扩展（目标3）。如果TG促进DM 1和CDM 1成肌细胞中突变型DMPK mRNA的降解， 和具有长扩增的成纤维细胞，GSK 3的抑制剂将代表靶向引发的小分子。 CDM 1和DM 1的原因。所获得的知识将是关键的进一步发展的临床试验 CDM 1和DM 1基于GSK 3 β的校正。

项目成果

Correction of Glycogen Synthase Kinase 3β in Myotonic Dystrophy 1 Reduces the Mutant RNA and Improves Postnatal Survival of DMSXL Mice.

强直性肌营养不良 1 中糖原合酶激酶 3β 的校正减少了突变 RNA 并提高了 DMSXL 小鼠的产后存活率。

• DOI: 10.1128/mcb.00155-19
• 发表时间: 2019
• 期刊: Molecular and cellular biology
• 影响因子: 5.3
• 作者: Wang,Mei;Weng,Wen-Chin;Stock,Lauren;Lindquist,Diana;Martinez,Ana;Gourdon,Genevieve;Timchenko,Nikolai;Snape,Mike;Timchenko,Lubov
• 通讯作者: Timchenko,Lubov