Enhancing Immunological Memory Using Aptamertargeted siRNA Delivery to T Cells

使用适体靶向 siRNA 递送至 T 细胞增强免疫记忆

基本信息

批准号：
9379074
负责人：
Eli Gilboa
金额：
$ 10.08万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-09 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9379074
关键词：
4T1 AXIN1 protein Address Affinity Antigens Antineoplastic Agents Applications Grants Biodistribution Blocking Antibodies Blood Breast Carcinoma CD8-Positive T-Lymphocytes Cancer Patient Cancer Vaccines Carcinogens Cell Differentiation process Cells Chemosensitization Chronic Clinical Clinical Trials Communicable Diseases Cytotoxic T-Lymphocyte-Associated Protein 4 Development Drug Kinetics Exhibits FRAP1 gene Generations Genetic Goals Hematopoietic Hematopoietic System Human Immune Immune response Immune system Immunity Immunologic Memory Immunologics In Vitro Investigation Ligands Malignant Neoplasms Mediating Mediator of activation protein Memory Modality Modeling Mus Oligonucleotides Operative Surgical Procedures Pathway interactions Patients Pharmaceutical Preparations Pharmacology Phenotype RNA Interference Raptors Sirolimus Small Interfering RNA T memory cell T-Lymphocyte Therapeutic Toxic effect Tumor Immunity Vaccines Validation antiviral immunity aptamer base clinical application cost effective effective therapy immunogenicity interest knock-down mTOR inhibition melanoma memory acquisition nonhuman primate novel pleiotropism prevent public health relevance response tool tumor uptake

项目摘要

DESCRIPTION (provided by applicant): Recent studies in mice, nonhuman primates, and clinical trials in human patients have emphasized the importance of the persistence of the vaccine-induced immune response, immunological memory, in mediating protective immunity against infectious diseases and cancer. Notably, inhibition of mediators of effector differentiatio like mTOR, GSK3b, Blimp-1 or T-bet, using genetic means or whenever available pharmacological agents, not only prevented the accumulation of the short-lived effectors but also redirected the activated T cells to differentiate along the memory pathway, and potentiated vaccine-induced protective immunity in mice. Notwithstanding, pharmacological agents, like rapamycin that was used to inhibit mTOR, often exhibit undesirable immune suppressive effects reflecting the broad distribution of their targets. Here we propose to develop a versatile, broadly applicable, and clinically feasible approach to promote the generation of memory T cell responses that addresses the main limitations of pharmacological agents. We propose to use RNAi to downregulate intracellular mediators of effector differentiation that will be targeted to CD8+ T cells by conjugation to oligonucleotide aptamer ligands. Aptamer and aptamer-siRNA conjugates offer potentially important advantages in terms of synthesis, conjugation, and reduced immunogenicity. The central hypothesis of the proposed studies is that aptamer-targeted siRNA inhibition of intracellular mediators in vaccine-induced CD8+ T cells will enhance their differentiation into long-lasting memory T cells and potentiate antitumor immunity that will be superior to pharmacological agents in terms of reduced toxicity, increased efficacy, and applicability to "nondrugable" intracellular targets. The proposed approach is supported by preliminary studies showing that 4-1BB aptamer-targeted raptor siRNA inhibition of mTORC1 function in activated CD8+ T cells led to the generation of a potent memory response and enhanced vaccine-induced protective immunity in tumor-bearing mice that was superior to that of rapamycin. The specific goal of the studies proposed in this application is to identify a best-i-class aptamer-siRNA conjugate to potentiate vaccine-induced protective immunity as determined in murine tumor models (Aims #1 and #2), that will guide the development of human conjugates capable of promoting the persistence of antigen- activated T cells in vitro (Aim #3). Successful accomplishment of the goals of the proposed studies will set the stage for clinical trials to potentiate vaccine-induced protective immunity in cancer patients using the agents developed in this proposal. Promoting memory differentiation by inhibition of intracellular mediators using siRNAs that are targeted to activated CD8+ T cells by conjugation to oligonucleotide aptamer ligands is arguably novel. The ability to target siRNAs to specific subsets of circulating immune or hematopoietic cells will provide a novel tool to manipulate the immune and hematopoietic systems for both investigational and therapeutic purposes.

描述（由申请人提供）：针对人类患者的小鼠，非人类灵长类动物和临床试验的最新研究强调了疫苗诱导的免疫反应，免疫记忆，免疫记忆的持续性在介导对感染性疾病和癌症的保护性免疫中的重要性。值得注意的是，使用遗传手段或使用可用的药理剂时，抑制效应介质分化的介体，例如MTOR，GSK3B，BLIMP-1或T-BET，不仅可以阻止短暂寿命效应子的积累，而且还将活化的T细胞重新定向以沿记忆途径和有效的疫苗诱导的保护型免疫机中的分化。尽管如此，药理学剂，例如用于抑制MTOR的雷帕霉素，通常会表现出不良的免疫抑制作用，反映出其靶标的广泛分布。在这里，我们建议开发一种通用的，广泛的适用且在临床上可行的方法来促进记忆T细胞反应的产生，以解决药理剂的主要局限性。我们建议使用RNAi下调效应子分化的细胞内介质，该介体将通过连接到寡核苷酸适体配体来靶向CD8+ T细胞。适体和适体 - 丝娜结合物在合成，结合和降低免疫原性方面具有潜在的重要优势。拟议的研究的中心假设是，适体靶向的siRNA抑制疫苗诱导的CD8+ T细胞中细胞内介质的抑制作用将增强其分化为长期持久的记忆T细胞，并增强抗肿瘤免疫力，并增强抗肿瘤免疫力，这将在毒性，效率上提高的效率和可效率的靶向性方面优于药理药物。提出的方法得到了初步研究的支持，表明4-1BB靶向的猛禽siRNA抑制MTORC1在活化的CD8+ T细胞中的功能，导致产生有效的记忆响应，并增强了疫苗诱导的抗瘤性抗蛋白酶的保护性免疫力。本应用中提出的研究的具体目标是确定最佳I级适体 - siRNA结合物，以增强疫苗诱导的保护性免疫，如鼠肿瘤模型（AIMS＃1和2），这将指导能够发展的人类偶联物的发展，能够促进抗原激活的TIT细胞的持久性，从而促进抗原激活细胞的效率（AIL AIM＃3）。成功实现拟议研究的目标将为使用本提案中开发的药物增强疫苗诱导的癌症患者的保护性免疫的临床试验奠定基础。通过使用siRNA抑制细胞内介质来促进记忆分化，这些siRNA通过与寡核苷酸适体配体共轭而针对激活的CD8+ T细胞来促进记忆。将siRNA靶向循环免疫或造血细胞的特定子集的能力将提供一种新颖的工具来操纵研究和治疗目的的免疫和造血系统。