Determination of optimal antisense oligonucleotide chemistry for efficient and safe splicing modulation in T cells

确定最佳反义寡核苷酸化学，以实现 T 细胞中高效、安全的剪接调节

基本信息

批准号：
9907140
负责人：
Gaddiel Galarza-Munoz
金额：
$ 28.37万
依托单位：
AUTOIMMUNITY BIOLOGIC SOLUTIONS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-05 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9907140
关键词：
Address Affect Animal Model Antisense Oligonucleotides Autoimmune Diseases Autoimmune Process Award Biological Biological Assay Callithrix Cell model Cells Chemicals Chemistry Clinic Clinical Research Cognitive Complex Cultured Cells Demyelinations Development Diagnosis Disease Emotional Etiology Exclusion Exhibits Exons Experimental Autoimmune Encephalomyelitis FDA approved Fruit Funding Future Generations Genetic Genetic Diseases Goals Grant Hepatic Hepatotoxicity Human IL7R gene Immune Immune System Diseases Immune system Immunologics Immunomodulators Immunosuppression Immunotherapy Insulin-Dependent Diabetes Mellitus Interleukin 7 Receptor Investigation Kidney Knowledge Lead Measures Mediating Medical Membrane Modeling Modification Molecular Multiple Sclerosis Neuraxis Neurologic Dysfunctions Neurons Pathogenicity Patients Pharmaceutical Preparations Pharmacology Phase Property Protein Isoforms RNA Splicing Research Rheumatoid Arthritis Safety Sensory Severity of illness Side Small Business Innovation Research Grant Small Business Technology Transfer Research Systemic Lupus Erythematosus T-Lymphocyte Testing Therapeutic Therapeutic Index Therapeutic Intervention Tissues Toxic effect Toxicology Translating Up-Regulation Work base cell type emerging adult immune function improved in vivo in vivo evaluation mRNA Precursor motor impairment mouse model nephrotoxicity nervous system disorder neuron loss novel personalized medicine preclinical study prevent side effect success targeted treatment tool uptake

项目摘要

PROJECT SUMMARY Multiple Sclerosis (MS) is the most common neurological disease of early adulthood and is mediated by autoimmune mechanisms that lead to demyelination and neuronal damage in the central nervous system, resulting in progressive neurological dysfunction. There is no cure for the disease and current treatments focus on preventing future immunological attacks, mainly by suppressing the immune system. This leads to adverse side effects that are often severe or fatal. Accordingly, there is a clear unmet need for the development of effective and well-tolerated therapies to arrest MS development. This has been challenging because MS has numerous etiologies and the molecular mechanisms underlying these etiologies are not well understood. We uncovered the molecular underpinnings of an MS etiology and hope this knowledge will translate into a targeted therapy for MS. This specific etiology results from up-regulation of the soluble form of the Interleukin- 7 Receptor (sIL7R), which has been shown to aggravate the progression and severity of the disease in the Experimental Autoimmune Encephalomyelitis (EAE) mouse model of MS, and to be elevated in patients of several autoimmune diseases including MS, Type I diabetes, Rheumatoid arthritis and Systemic lupus erythematosus. Given that sIL7R is produced by abnormal exclusion of exon 6 from IL7R pre-mRNAs, we developed a novel biologic drug, a splicing-modulating antisense oligonucleotide (SM-ASO; IL7R-005) that corrects this abnormal splicing and restores normal expression of IL7R protein isoforms. IL7R-005 represents a major improvement over current MS therapies in that by correcting IL7R splicing, it diminishes expression of the pathogenic sIL7R isoform, without affecting the function of the membrane-bound IL7R (mIL7R), which is vital for proper immune function, thereby avoiding the adverse immunosuppressive effects of current drugs. T cells are the major producers of sIL7R in humans, and thus to reduce sIL7R levels, IL7R-005 needs to be delivered into T cells in vivo. Although SM-ASOs have been shown to modulate RNA splicing decisions in many tissues in vivo (e.g. FDA-approved Spinraza), the delivery and functionality of SM-ASOs in T cells have not been thoroughly examined, and represents the major hurdle to expand the use of SM-ASOs for treatment of immunological disorders and the development of novel immunotherapies. Here, we address this obstacle by conducting an in-depth, side-by-side analysis of the influence of diverse chemical modifications on the efficiency of SM-ASOs in primary T cells and their potential toxic effects in relevant cell models. This is critical as the chemical modifications of the SM-ASOs could influence their pharmacological properties (e.g., cellular uptake) differentially across cell-types, and thus define the potency of SM-ASOs in a cell-type specific matter. The chemical modifications of the SM-ASOs also dictate potential harmful effects, such as hepatic or renal toxicities. Therefore, this in-depth analysis will enable selection of the chemistry with the optimal therapeutic index (i.e., high potency, low toxicity) for efficient splicing modulation in T cells.

项目摘要多发性硬化症（MS）是成年早期最常见的神经系统疾病，由自身免疫机制导致中枢神经系统中脱髓鞘和神经元损害，导致进行性神经功能障碍。无法治愈该疾病，目前的治疗重点在预防未来的免疫攻击方面，主要是通过抑制免疫系统。这导致不利副作用通常是严重或致命的。因此，对于开发的明显未满足有效且耐受良好的疗法可阻止MS开发。这很具有挑战性，因为MS有这些病因的许多病因和分子机制尚不清楚。我们发现了MS病因的分子基础，并希望这些知识将转化为针对MS的靶向疗法。这种特定的病因是由白介素 - 可溶性形式的上调引起的 7受体（SIL7R），已证明会加剧该疾病在疾病中的进展和严重程度 MS的实验性自身免疫性脑脊髓炎（EAE）小鼠模型，并在患者中升高几种自身免疫性疾病，包括MS，I型糖尿病，类风湿关节炎和全身性狼疮红斑。鉴于SIL7R是通过异常排除外显子6从IL7R PREMRNA中产生的，我们开发了一种新型的生物学药物，一种剪接调节反义寡核苷酸（SM-ASO; IL7R-005）纠正这种异常剪接并恢复IL7R蛋白同工型的正常表达。 IL7R-005代表对当前MS疗法的重大改进是通过校正IL7R剪接，它减少了致病SIL7R同工型，不影响膜结合的IL7R（MIL7R）的功能，这是对于适当的免疫功能至关重要，从而避免了当前药物的不良免疫抑制作用。 t 细胞是人类SIL7R的主要生产国，因此为了降低SIL7R水平，IL7R-005必须为在体内递送到T细胞中。尽管已显示SM-ASO在调节RNA剪接决策中许多组织在体内（例如FDA批准的Spinraza），T细胞中SM-ASO的传递和功能具有未经彻底检查，代表扩大SM-ASO用于治疗的主要障碍免疫障碍和新型免疫疗法的发展。在这里，我们通过对各种化学修改对该影响的影响进行深入的，并排分析原代T细胞中SM-ASO的效率及其在相关细胞模型中的潜在毒性作用。这很关键由于SM-ASO的化学修饰可能影响其药理特性（例如，细胞跨细胞类型的摄取），从而定义了细胞类型特定物质中SM-ASO的效力。 SM-ASO的化学修饰还决定了潜在的有害作用，例如肝或肾脏毒性。因此，这种深入的分析将能够通过最佳治疗方法选择化学指数（即高效力，低毒性），用于T细胞中有效的剪接调制。