Strategy for specific delivery of antisense oligonucleotides to T cells

将反义寡核苷酸特异性递送至 T 细胞的策略

• 批准号: 10547347
• 负责人: Gaddiel Galarza-Munoz
• 金额: $ 30.65万
• 依托单位: AUTOIMMUNITY BIOLOGIC SOLUTIONS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-17 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10547347
• 关键词: Address; Antibodies; Antisense Oligonucleotides; Autoimmune Diseases; Autoimmunity; Binding; Biological; CD4 Positive T Lymphocytes; Cell Surface Receptors; Cell physiology; Cells; Cellular biology; Clinical; Communicable Diseases; Development; Disease; Dose; Ensure; Exclusion; Exons; Eye; Gene Expression; Genes; Genetic Diseases; Goals; Health; Hepatic; Hepatocyte; Homeostasis; Human; IL2RB gene; Immune; Immunologic Deficiency Syndromes; Immunologics; Immunooncology; Interleukin 2 Receptor; Interleukin 7 Receptor; Liver; Maintenance; Malignant Neoplasms; Mediating; Methods; Monoclonal Antibodies; Mus; Muscle; Nerve Degeneration; Neuraxis; Peptides; Pharmaceutical Preparations; Pharmacotherapy; Phase; Proteins; Proteome; RNA Splicing; Role; Safety; Specificity; Spleen; T-Cell Immunologic Specificity; T-Lymphocyte; Testing; Therapeutic; Therapeutic Index; Thymus Gland; Tissues; Toxic effect; Transfection; Tumor Suppression; Vaccines; Validation; Visualization; Work; antibody conjugate; base; cancer cell; cancer therapy; cell type; empowered; fluorophore; immunoregulation; improved; in vivo; in vivo Model; lymph nodes; nanobodies; nonhuman primate; peripheral blood; programmed cell death protein 1; receptor; receptor mediated endocytosis; response; success; tool; tumor; uptake; vaccine development; vaccine response

PROJECT SUMMARY Antisense oligonucleotides (ASOs) are promising drugs given their potential to modify expression of disease- related genes, including those previously considered ‘undruggable’. Despite their clinical potential, their success has been limited to hepatic, muscular and neurodegenerative conditions due to limitations with delivering ASOs to tissues other than liver, muscle and the central nervous system (CNS), which are tissues with either rich distribution of ASOs or specific delivery methods. Unfortunately, applications to immunological conditions or conditions that could benefit from immuno-modulation (e.g., cancer, vaccines) have been limited due to the lack of effective tools to deliver ASOs to immune cells in vivo, in particular to T cells. Accordingly, delivery tools that could enhance the efficiency and specificity of ASO delivery to T cells remain a critical unmet need. To address this critical need, ABS is developing a modular delivery platform to target ASOs specifically to T cells utilizing conjugated mono-specific monoclonal antibodies (mmAbs) against T cell-specific cell surface receptors. mmAbs are highly specific antibodies that have been screened against the human proteome and selected for their exclusive binding to the target protein. Such selectivity screen is not usually employed in the development of monoclonal antibodies, resulting in antibodies that bind to other proteins besides their intended target, thereby enhancing potentially toxic effects due to off-target binding. Accordingly, the mmAbs proposed here have the potential to enhance delivery of ASOs to T cells while reducing potential toxic effects of the antibody conjugate. ABS’ T cell-specific delivery platform takes advantage of the T cell-predominant expression of the Programmed Cell Death Protein 1 (PDCD1, PD1, CD279) and Interleukin 2 Receptor Subunit Beta (IL2RB, CD122). Besides their T cell-predominant expression, their biological roles in T cells make them attractive candidates. PDCD1 is highly expressed in tumor-infiltrating T cells and mediates suppression of these tumor-reactive T cells, thereby hindering their efficacy to kill cancer cells. Accordingly, -PDCD1 antibodies could have a dual role in enhancing tumor reactivity by directing immuno-modulatory ASOs to these critical cells, while also relieving the PDCD1- mediated suppression of tumor-reactive T cells. IL2RB promotes receptor-mediated endocytosis in T cells, and here we leverage this function to target and drive internalization of the conjugated ASO specifically in T cells. The goal of this Phase I proposal is to test the efficiency and specificity of -PDCD1 and -IL2RB mmAbs to deliver the conjugated ASOs to T cells ex vivo and in mice. Successful completion of this goal will validate the utility of ABS’ T cell-specific delivery platform to deliver ASOs to T cells in vivo. This modular delivery platform will have broad impact on human health given its wide applicability for treatment of cancers, immunodeficiencies, autoimmune and infectious diseases, and applications in vaccine development. In the ensuing Phase II proposal, we will create a mono-specific nanobody of the optimal mmAb to further improve its therapeutic index, and test its efficacy and safety to deliver ASOs to T cells in mice and nonhuman primates.

项目摘要 反义寡核苷酸（ASO）是有前途的药物，因为它们有可能改变疾病表达 相关基因，包括先前被认为是“不可能”的基因。尽管具有临床潜力，但他们的成功 由于限制了ASOS 到肝脏，肌肉和中枢神经系统（CNS）以外的组织，它们是富含的组织 ASO或特定递送方法的分布。不幸的是，应用于免疫条件或 由于缺乏，可能受益于免疫调节（例如癌症，疫苗）的疾病因缺乏而受到限制 在体内向免疫细胞（尤其是T细胞）传递ASO的有效工具。彼此之间，交付工具 可以提高ASO向T细胞递送的效率和特异性仍然是一个关键的未满足需求。 为了满足这一关键需求，ABS正在开发一个模块化递送平台，以专门针对T细胞的ASO 利用针对T细胞特异性细胞表面受体的共轭单特异性单克隆抗体（MMAB）。 MMAB是已针对人蛋白质组筛选并选择的高度特异性抗体 它们与靶蛋白的独家结合。这种选择性屏幕通常不在开发中进行 单克隆抗体的抗体，导致与其他蛋白质结合的抗体，除了其预期靶标之外 由于脱靶结合而增加了潜在的毒性作用。根据，这里提议的MMAB有 增强ASO向T细胞递送的潜力，同时减少抗体结合物的潜在毒性作用。 ABS的T细胞特异性输送平台利用了程序的T细胞促销表达 细胞死亡蛋白1（PDCD1，PD1，CD279）和白介素2受体亚基β（IL2RB，CD122）。除了 他们的T细胞促进表达，它们在T细胞中的生物学作用使其具有吸引力的候选者。 PDCD1是 在肿瘤浸润的T细胞中高度表达并介导这些肿瘤反应性T细胞的抑制作用，从而 阻碍其杀死癌细胞的有效性。彼此之间，-PDCD1抗体可以在增强中具有双重作用 肿瘤反应性通过将免疫调节性ASO引入这些关键细胞，同时还可以缓解PDCD1-- 介导的抑制肿瘤反应性T细胞。 IL2RB促进受体介导的T细胞中的内吞作用，并促进 在这里，我们利用此功能来靶向和驱动集成ASO专门在T细胞中的内在化。 该阶段I建议的目的是测试-PDCD1和-IL2RB MMAB的效率和特异性 将连接的ASO传递给T细胞离体和小鼠。成功完成此目标将验证 ABS的T细胞特异性输送平台的实用性，可在体内向T细胞传递ASO。这个模块化交付平台 鉴于人类对癌症的治疗，免疫缺陷，将对人类健康产生广泛的影响 自身免疫性和传染病，以及疫苗开发中的应用。在随后的第二阶段提案中， 我们将创建一个最佳MMAB的单特异性纳米机构，以进一步改善其治疗指数并进行测试 它可以在小鼠和非人类隐私的T细胞中传递ASO的有效性和安全性。