Development of novel ASO-based therapeutics for CMT1A

开发基于 ASO 的 CMT1A 新型疗法

• 批准号: 10383878
• 负责人: Steve OConnor
• 金额: $ 25.66万
• 依托单位: SHIFT PHARMACEUTICALS HOLDINGS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10383878
• 关键词: 3' Untranslated Regions; Acute; Alzheimer' s Disease; Animals; Antisense Oligonucleotides; Biological Monitoring; Cell Culture Techniques; Cells; Characteristics; Charcot-Marie-Tooth Disease; Chemistry; Clinic; Clinical; Development; Disease; Disease Progression; Disease model; Dose; Electrophysiology (science); Enhancers; Exons; Fire - disasters; Future; Gene Targeting; Genes; Genetic Diseases; Genetic Transcription; Genetically Engineered Mouse; Genomics; Grant; Human; Human Cell Line; In Vitro; Injections; Intraperitoneal Injections; K-562; Knock-out; Lead; Length; Mediating; Messenger RNA; Missouri; Modeling; Molecular; Molecular Genetics; Monitor; Mus; Muscle; Nerve; Nerve Tissue; Nucleic Acids; Oligonucleotides; PMP22 gene; Pathway interactions; Patients; Peripheral Nerves; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Production; Proteins; Protocols documentation; RNA Splicing; Rare Diseases; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Risk; SMN protein (spinal muscular atrophy); Safety; Schwann Cells; Series; Signal Transduction; Site; Stretching; Structure; Subcutaneous Injections; Symptoms; Syndrome; Techniques; Testing; Therapeutic; Tissue Extracts; Translating; Translation Process; Translational Repression; Translations; Universities; Vertebral column; Work; Yeast Artificial Chromosome; base; cohort; design; dosage; drug candidate; efficacy evaluation; efficacy study; exon skipping; experimental study; humanized mouse; in vivo; lead candidate; loss of function; mRNA Expression; mRNA Precursor; mRNA Transcript Degradation; mouse model; novel; nuclease; nucleic acid-based therapeutics; overexpression; phosphorodiamidate morpholino oligomer; precision medicine; prevent; programs; safety study; skills; targeted treatment

Abstract: The objective of this project is to develop a safe, effective antisense oligonucleotide with Phosphorodiamidate Morpholino Oligomer (PMO) backbone chemistry capable of significantly reducing PMP22 protein production in Schwann cells, which is titratable in its dosing characteristics. For patients with CMT1A, the most common form of CMT, PMP22 over production of this protein drives disease progression. We have previously performed rigorous analysis of the PMP22 gene sequence, which has led to the design of numerous ASOs that have been shown to effectively alter PMP22 mRNA production. These PMOs have been designed to alter exon splicing, resulting in an “exon-skipped” product that is severely truncated early in the translation process, effectively reducing PMP22 protein production. Importantly, however, PMP22 production is not completely eliminated since this would also be detrimental. CMT1A is a remarkably common rare disease that is caused by the overexpression of a protein involved in the function of nerves in the periphery. The protein, PMP22, acts much like an insulator, allowing nerves to fire their signal properly. While too much of PMP22 is bad for the cell, too little of PMP22 is also a problem and therefore any therapeutic strategy cannot simply knock-out the expression of the important gene. The molecular genetics of CMT1A makes this disease particularly amenable for nucleic acid-based therapeutics designed to modulate PMP22 expression since: 1) CMT1A is monogenic; 2) the disease gene has been identified; and 3) inhibition of PMP22 expression can be accomplished through a variety a molecular mechanisms. Currently, no drugs are available for effectively treating the disease progression of CMT1A, thus this highly at risk patient class remains untreated with disease altering therapies. During Phase 1, a number of the PMO ASO candidates will be screened “in vivo”, in a humanized PMP22 mouse model for activity and acute safety. In Aim 1, we will inject selected compounds previously screened in cell based models into mice and monitor the PMP22 mRNA total for full-length and “exon-skipped” mRNA production. in Schwan cells surrounding peripheral nerve tissues (after sacrifice) to confirm efficacy and monitor overall in vivo efficiency. Once active compounds are confirmed, in Aim 2 we will inject molecules into a different cohort of mice and monitor biological activity of nerve function using accepted electrophysiology techniques. Previously, Shift Pharmaceuticals (collaborating with the University of Missouri) has successfully demonstrated the PMO ASO approach for altering exon splicing with another drug candidate (E1v1.11) that has proven to be extremely effective at modifying protein production of SMN protein for Spinal Muscular Atrophy disease, which suggested this approach should be effective in mouse models for CMT. We have also demonstrated in previous studies that E1v1.11 and similar PMOs are very safe when dosed at extremely high levels, indicating that future safety studies of molecules developed for CMT should also demonstrated acceptable safety profiles.

抽象的： 该项目的目的是用磷化二次的安全有效，有效的反义寡核苷酸 形态寡聚（PMO）骨干化学能够显着降低PMP22蛋白质的产生 Schwann细胞，其给药特征是钛。对于CMT1A的患者，最常见的形式 CMT的PMP22过于生产该蛋白质会驱动疾病进展。我们以前已经表演了 对PMP22基因序列的严格分析，这导致了许多ASO的设计 显示可有效改变PMP22 mRNA的产生。这些PMO已设计为改变外显子剪接， 导致“外显子甲组合”产品在翻译过程中被严重截断，有效地被严重截断 减少PMP22蛋白质的产生。但是，重要的是，PMP22的生产尚未完全消除，因为 这也是有害的。 CMT1A是一种非常常见的罕见疾病，是由参与该蛋白质的过表达引起的 神经在周围的功能。蛋白质PMP22的作用很像绝缘体，使神经发射 正确信号。虽然PMP22的太多对单元不利，但PMP22的太少也是一个问题，因此 任何理论策略都不能简单地淘汰重要基因的表达。分子遗传学 CMT1A的疾病特别适合基于核酸的治疗疗法 PMP22表达以来：1）CMT1A是单基因的； 2）疾病基因已被鉴定出来； 3）抑制 PMP22表达可以通过多种分子机制来完成。目前，没有毒品 可用于有效治疗CMT1A的疾病进展 未经疾病改变疗法的治疗。 在第1阶段，将在人源化的PMP22小鼠中筛选许多PMO ASO候选者。 活动和急性安全的模型。在AIM 1中，我们将注入以前在基于单元的基于细胞的化合物 在小鼠中模型并监测PMP22 mRNA总数的全长和“外显子” mRNA产生。 周围周围神经组织周围的雪旺细胞（牺牲后）以确认效率并监测整体体内 效率。一旦确认活性化合物，在AIM 2中，我们将分子注入不同的小鼠队列 并使用公认的电生理技术来监测神经功能的生物学活性。 以前，Shift Pharmaceuticals（与密苏里大学合作）已成功证明 与另一位候选药物（E1V1.11）更改外显子剪接的PMO ASO方法已被证明为 在修饰SMN蛋白的蛋白质产生的脊柱肌肉萎缩疾病方面极为有效， 建议这种方法在CMT的小鼠模型中应有效。我们也在以前的 研究E1V1.11和类似PMO在极高的水平时非常安全，表明未来 针对CMT开发的分子的安全研究也应证明可接受的安全性概况。