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.

项目概要多发性硬化症 (MS) 是成年早期最常见的神经系统疾病，由导致中枢神经系统脱髓鞘和神经元损伤的自身免疫机制，导致进行性神经功能障碍。该疾病无法治愈，目前的治疗重点主要通过抑制免疫系统来预防未来的免疫攻击。这会导致不利的副作用通常是严重或致命的。因此，显然存在未满足的发展需求有效且耐受性良好的疗法可阻止多发性硬化症的发展。这一直具有挑战性，因为 MS 已经许多病因学以及这些病因学背后的分子机制尚不清楚。我们揭示了多发性硬化症病因学的分子基础，并希望这些知识能够转化为 MS 的靶向治疗。这种特定的病因是由于可溶性白细胞介素的上调所致。 7 受体 (sIL7R)，已被证明会加剧疾病的进展和严重程度多发性硬化症的实验性自身免疫性脑脊髓炎 (EAE) 小鼠模型，并在以下患者中升高多种自身免疫性疾病，包括多发性硬化症、I 型糖尿病、类风湿性关节炎和系统性狼疮红斑狼疮。鉴于 sIL7R 是通过从 IL7R 前体 mRNA 中异常排除外显子 6 产生的，我们开发了一种新型生物药物，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 蛋白亚型的正常表达。 IL7R-005代表与目前的多发性硬化症疗法相比，这是一个重大改进，通过纠正 IL7R 剪接，它可以减少致病性 sIL7R 亚型，而不影响膜结合 IL7R (mIL7R) 的功能，即对于正常的免疫功能至关重要，从而避免当前药物的不良免疫抑制作用。时间细胞是人类 sIL7R 的主要生产者，因此为了降低 sIL7R 水平，需要将 IL7R-005 递送至体内 T 细胞。尽管 SM-ASO 已被证明可以调节 RNA 剪接决策 SM-ASO 在体内的许多组织中（例如 FDA 批准的 Spinraza），T 细胞中 SM-ASO 的递送和功能尚未经过彻底检查，是扩大 SM-ASO 治疗用途的主要障碍免疫疾病的研究和新型免疫疗法的开发。在这里，我们通过以下方式解决这个障碍对各种化学修饰对 SM-ASO 在原代 T 细胞中的效率及其在相关细胞模型中的潜在毒性作用。这很关键因为 SM-ASO 的化学修饰可能会影响其药理学特性（例如，细胞吸收）在细胞类型之间存在差异，从而定义了 SM-ASO 在细胞类型特定物质中的效力。 SM-ASO 的化学修饰也决定了潜在的有害影响，例如肝或肾毒性。因此，这种深入的分析将能够选择具有最佳治疗效果的化学物质 T 细胞中有效剪接调节的指数（即高效、低毒）。