Restoring Vision with High-Fidelity Nonsense Codon Correction

通过高保真无义密码子校正恢复视力

基本信息

批准号：
10156779
负责人：
Christopher A Ahern
金额：
$ 145.48万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10156779
关键词：
Accounting Address Amino Acids Anatomy Anticodon Biochemical Biological Assay Biological Availability Biological Models Blindness Cell Differentiation process Cell membrane Cells Childhood Clinical Trials Communication Communities Complex Cone dystrophy Cultured Cells DNA DNA Sequence Data Defect Development Disease Drug Delivery Systems Drug Targeting Electrophysiology (science)Engineering Evaluation Eye FDA approved Gene Targeting Genes Genetic Diseases Glutamine Human In Vitro Incidence Inherited Ion Channel Ion Channel Protein Iowa Libraries Light Location Membrane Proteins Methods Missense Mutation Molecular Monitor Mus Mutation Neural Retina Night Blindness Nonsense Codon Nonsense Mutation Nucleic Acids Nucleotides Ophthalmologist Organoids Pathogenesis Patients Pharmaceutical Preparations Pharmacology Photoreceptors Phototransduction Physiology Point Mutation Post-Translational Protein Processing Production Protein Biosynthesis Protein translocation Proteins Proteomics Research Research Personnel Resources Retina Retinal Diseases Retinal Photoreceptors Ribosomes Safety Scientist Silicon Dioxide Site Structure Structure of retinal pigment epithelium System TRPM1 gene Technology Terminator Codon Testing Therapeutic Therapeutic Intervention Tissues Toxic effect Transfer RNA Translations Universities Viral Vitelliform macular dystrophy base cell type disease phenotype efficacious treatment functional outcomes gene therapy genome editing human disease human pluripotent stem cell in vivo induced pluripotent stem cell intravitreal injection mouse model nanomaterials nanomedicine nanoparticle new technology next generation nonhuman primate novel polypeptide pre-clinical pre-clinical therapy preclinical safety premature repaired retinal progenitor cell safety assessment safety testing sight restoration small molecule small molecule therapeutics stem cell biology tRNA Precursor targeted delivery therapeutic gene therapeutic target transcriptomics virtual

项目摘要

PROJECT SUMMARY/ABSTRACT Nonsense mutations cause approximately 15% of genetically inherited retinopathies and inherited human diseases in general, accounting for 2.5 to 3 million patients in the U.S. For certain specific genes, nonsense mutation incidences can be as high as 40%. Because nonsense mutations cause premature termination (PTC) of protein translation, the disease phenotype is often severe. Currently, there are only a limited number of therapies for nonsense mutations being tested in human clinical trials, including gene therapy, small molecule read-through drugs, or genome editing. Associated challenges equal the promises of each of these therapeutic options. Looking forward, newer technologies may address these hurdles and provide more safe and efficacious treatments for patients. During protein translation, tRNA functions at the ribosomal site to incorporate a specific amino acid into the polypeptide sequence. We aim to develop the next generation of nucleic acid therapy based on anticodon encoding transfer RNA (ace-tRNA) that incorporates the correct wild type amino acid at the site of a disease-causing nonsense mutation. Because of the many anatomical advantages afforded by the eye, we seek to test the broad applicability of ace-tRNA therapeutics for nonsense mutations that cause retinopathies and related blindness due to defects in a variety of genes, including those encoding ion channel proteins. Specifically we will focus on nonsense mutation in ion channels expressed in photoreceptors (PR) which convert retinal light inputs and retinal pigment epithelium (RPE), which provide support for PR. These two cell types are primarily the site of blindness pathogenesis. In this project, we will: 1) Develop ace-tRNA therapeutics that target specific nonsense mutations across several PR and RPE ion channels. 2) Engineer both viral and non-viral ace-tRNA delivery systems for long-term editing. Using these we will determine the functional outcome of ace-tRNA treatment using cultured cells and human iPSC-derived RPE and iPSC-PR retinal organoids. 3) Test both our viral and non-viral ace-tRNA in vivo using mice harboring genetic defects that cause blindness in humans; and 4) Assess the safety and bioavailability of ace-tRNA therapeutics in our preclinical NHP model systems. There are no FDA-approved therapeutic drugs that target channelopathies because of the complexities associated with precise post-translational modifications, carefully regulated expression, and assembly. Our team’s combined expertise in ace-tRNA development, nanomaterial synthesis, human pluripotent stem cell biology, ion-channel physiology, and pathophysiological model systems is unique and ideally suited to advance ace-tRNA technology toward clinical trials for a wide range of genetic diseases that cause blindness.

项目概要/摘要无义突变导致约 15% 的遗传性视网膜病和遗传性人类视网膜病一般疾病，美国有 2.5 至 300 万患者。对于某些特定基因来说，这是无稽之谈突变发生率可高达 40%，因为无义突变会导致过早终止 (PTC)。蛋白质翻译的疾病表型往往很严重，目前只有有限的数量。正在人体临床试验中测试无义突变的疗法，包括基因疗法、小分子疗法通读药物或基因组编辑的相关挑战等于这些疗法的前景。展望未来，更新的技术可能会解决这些障碍并提供更安全和有效的选择。在蛋白质翻译过程中，tRNA 在核糖体位点发挥作用，整合特定的蛋白质。我们的目标是开发下一代基于核酸的疗法。编码转移 RNA (ace-tRNA) 的反密码子在位点处掺入了正确的野生型氨基酸由于眼睛具有许多解剖学优势，我们寻求测试 ace-tRNA 疗法对引起视网膜病的无义突变的广泛适用性以及由于多种基因（包括编码离子通道蛋白的基因）缺陷导致的相关失明。具体来说，我们将重点关注光感受器（PR）中表达的离子通道中的无义突变，这些突变将视网膜光输入和视网膜色素上皮 (RPE) 为 PR 提供支持。主要是失明发病部位。在这个项目中，我们将： 1) 开发针对多个 PR 和 RPE 离子的特定无义突变的 ace-tRNA 疗法渠道。 2) 设计病毒和非病毒 ace-tRNA 传递系统以进行长期编辑。使用培养细胞和人类 iPSC 衍生细胞确定 ace-tRNA 治疗的功能结果 RPE 和 iPSC-PR 视网膜类器官。 3) 使用携带遗传缺陷的小鼠体内测试我们的病毒和非病毒 ace-tRNA 人类失明；以及 4) 在我们的临床前 NHP 模型系统中评估 ace-tRNA 疗法的安全性和生物利用度。由于通道病的复杂性，目前尚无 FDA 批准的针对通道病的治疗药物与精确的翻译后修饰、仔细调节的表达和组装相关。团队在 ace-tRNA 开发、纳米材料合成、人类多能干细胞方面的综合专业知识生物学、离子通道生理学和病理生理学模型系统是独特的，非常适合推进 ace-tRNA 技术针对多种导致失明的遗传性疾病进行临床试验。