A Novel Class of Synthetic Receptors to Empower the Age of mRNA Therapies

一类新型合成受体将推动 mRNA 治疗时代的到来

基本信息

批准号：
10687517
负责人：
Xiaojing J Gao
金额：
$ 135.08万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-19 至 2026-09-18
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10687517
关键词：
Ablation Adoptive Cell Transfers Adverse effects Age Antigens Basic Science CAR T cell therapy Cell Extracts Cell physiology Cells Computer Simulation Cues DNA Dimerization Disease Dopamine Engineering Event Exposure to Fibroblasts General Population Genome Half-Life Immune Ligands Logic Logistics Malignant Neoplasms Mediating Messenger RNA Modality Mutation Myocardial Infarction Neurons Output Patients Population Precision therapeutics Process Proteins Public Health RNA Research Personnel Risk Safety Signal Transduction Surface T-Lymphocyte Tissue Engineering Transcript Vaccination Vaccines cellular engineering chimeric antigen receptor chimeric antigen receptor T cells coronavirus disease cost cytokine delivery vehicle design empowerment extracellular immunoengineering improved in vivo mRNA delivery manufacture notch protein novel novel therapeutics receptor response senescence tool vaccine efficacy

Ablation Adoptive Cell Transfers Adverse effects Age Antigens Basic Science CAR T cell therapy Cell Extracts Cell physiology Cells Computer Simulation Cues DNA Dimerization Disease Dopamine Engineering Event Exposure to Fibroblasts General Population Genome Half-Life Immune Ligands Logic Logistics Malignant Neoplasms Mediating Messenger RNA Modality Mutation Myocardial Infarction Neurons Output Patients Population Precision therapeutics Process Proteins Public Health RNA Research Personnel Risk Safety Signal Transduction Surface T-Lymphocyte Tissue Engineering Transcript Vaccination Vaccines cellular engineering chimeric antigen receptor chimeric antigen receptor T cells coronavirus disease cost cytokine delivery vehicle design empowerment extracellular immunoengineering improved in vivo mRNA delivery manufacture notch protein novel novel therapeutics receptor response senescence tool vaccine efficacy

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项目摘要

Abstract The COVID vaccination campaigns have highlighted the promise of mRNA-mediated delivery as a novel therapeutic modality. One particular application is adoptive cell therapy, where immune cells are equipped with novel functions to treat diseases, e.g., expressing chimeric antigen receptors (CARs) in T cells to ablate cancer and other undesirable cells. Compared to DNA-based engineering of immune cells, mRNA has several advantages as a delivery vector, especially its superior safety profile, because it eliminates the risk of randomly inserting into the host genome and causing mutations, and its short half-life mitigates long-term adverse effects due to the persistence of the engineered cellular function. Instead of extracting cells from the patient, engineering them ex vivo, and then reinducing them, researchers have even directly delivered CAR-encoding mRNAs and created functional CAR T cells in vivo. This is appealing because it has the potential to make adoptive cell therapy accessible to the general public, its logistics almost as straightforward as manufacturing, distributing, and administering vaccine shots, in contrast to the costly ex vivo engineering process that will be limited to the privileged few. Despite the great potential of mRNA-mediated adoptive cell therapy, there remains a critical need for tools that enhance targeting precision. It is very rare for a single surface marker, targeted by CAR, to unambiguously identify one target cell population. Therefore, “on-target/off-caner” killing is a major concern for CAR T cell therapies against cancer, and the same concern also applies to scenarios of ablating other cells, such as active fibroblasts in heart infarction or senescent cells. One elegant solution is synthetic receptors, most notably synthetic Notch, that detect a second marker and express CAR in response, effectively forming AND logic, where target cells are only killed when both inputs to the synthetic receptor and CAR are present. However, to our knowledge, all existing synthetic modular, programmable receptors operate at the DNA level, and are therefore incompatible with mRNA-mediated delivery. Here we design and demonstrate the feasibility of a first-in-class synthetic modular receptor that operate at the RNA level. It converts ligand-induced dimerization events into the expression of arbitrary output proteins. Through extensive computational simulation and experimental optimization, we will expand the input/output repertoire of this receptor, establish its design principle, enable its encoding on single transcripts and delivery by mRNA, and take first steps towards improving the precision of ablating active fibroblasts to treat heart infarction. The impact of such novel receptors is beyond adoptive cell therapy. For example, they can facilitate basic research by recording cells’ (e.g., neurons) exposure to specific signals (e.g., dopamine). They will benefit a variety of other biomedical applications too, from expressing antigens or cytokines in response to extracellular cues to enhance vaccine efficacy, to generating novel sense-and-respond capabilities for tissue engineering.

抽象的 COVID疫苗活动强调了mRNA介导的分娩的希望治疗方式。一种特殊应用是自适应细胞疗法，其中免疫细胞配备了治疗疾病的新功能，例如，在T细胞中表达嵌合抗原受体（CAR）以消融癌症和其他不良细胞。与免疫细胞基于DNA的工程相比，mRNA具有多个作为交付向量的优势，尤其是其出色的安全性，因为它消除了随机的风险插入宿主基因组并引起突变，其短期寿命减轻了长期不良反应由于工程细胞功能的持续存在。与其从患者中提取细胞，而是工程他们是体内的，然后重新诱导了它们，研究人员甚至直接交付了汽车编码的mRNA和在体内创建功能性汽车T细胞。这是因为它有可能制作自适应细胞的潜力公众可以使用的治疗，其物流几乎与制造，分发，并管理疫苗射击，与代价高昂的体内工程过程相反，该过程仅限于特权很少。尽管mRNA介导的自适应细胞疗法具有很大的潜力，但仍然需要增强定位精度的工具。对于由汽车靶向的单个表面标记非常罕见明确识别一个目标细胞群。因此，“靶标/脱离驾驶员”杀戮是一个主要关注的问题针对癌症的CAR T细胞疗法，同样的关注也适用于消灭其他细胞的场景，例如心脏梗塞或感觉细胞中的活性成纤维细胞。一种优雅的解决方案是合成受体，大多数值得注意的是合成缺口，检测第二个标记并在响应中表达汽车，有效地形成和逻辑，只有在存在合成接收器和汽车的两个输入时，目标细胞才会被杀死。然而，据我们所知，所有现有的合成模块化，可编程接收器均在DNA级别运行，并且是因此与mRNA介导的递送不兼容。在这里，我们设计并演示了操作的第一类合成模块化接收器的可行性在RNA级别。它将配体诱导的二聚事件转换为任意输出蛋白的表达。通过大量的计算模拟和实验优化，我们将扩展输入/输出该受体的曲目，建立其设计原理，使其在单个成绩单上进行编码，并通过 mRNA，并迈出了提高减轻活性成纤维细胞以治疗心脏梗塞的精确度的第一步。这种新型受体的影响超出了自适应细胞疗法。例如，它们可以促进基本通过记录细胞（例如神经元）暴露于特定信号（例如多巴胺）的研究。他们将受益从表达抗原或细胞因子的各种生物医学应用也是细胞外的提高疫苗效率的提示，从而为组织工程产生新颖的感官和响应能力。