Engineer Immune Cells via Chemoenzymatic Glycan Editing

通过化学酶聚糖编辑工程免疫细胞

基本信息

批准号：
10578671
负责人：
John Ross Teijaro
金额：
$ 94.43万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10578671
关键词：
Adoptive Cell Transfers Antibodies Antigens BLR1 gene Biological Process Biopolymers Biotin Biotinylation CD8-Positive T-Lymphocytes Cell Communication Cell Line Cell Therapy Cell surface Cells Chronic Chronic Lymphocytic Leukemia Dendritic Cells Development Embryonic Development Endowment Engineering Enzymes Flow Cytometry Fucose Fucosyltransferase Funding Glycocalyx Goals Helicobacter pylori Hematologic Neoplasms Immune Immune System Diseases Immune response Immunoglobulin Fragments Label Lymphoma Malignant Neoplasms Mediating Membrane Methods Modeling Monoclonal Antibodies Mus Natural Killer Cells Neoplasm Metastasis Polysaccharides Process Signal Transduction Specificity T-Lymphocyte Tumor Antigens Tumor Immunity Viral Cancer Virus Diseases anti-PD-1/PD-L1 anti-PD-L1 anti-PD-L1 therapy cell type chimeric antibody chimeric antigen receptor T cells cytotoxic disorder prevention exhaust exhaustion galactoside 3-fucosyltransferase glycosyltransferase human disease immune activation immunoengineering in vivo interest migration mouse model nanobodies neurotransmission success sugar sugar nucleotide tool tumor tumor eradication

项目摘要

PROJECT SUMMARY/ABSTRACT Cell-cell interactions are involved in numerous biological processes, including immune responses, embryonic development, neuronal signaling and cancer metastasis. Some major gaps in our understanding of cell-cell interactions include: what are the interacting partners of a specific cell type at different stages of a biological process? How to modify the cell-surface landscape of these interacting partners to tune the functions of a specific cell type with minimum perturbation of its endogenous functions? The objective of this project is to develop glycosyltransferase-based chemoenzymatic methods to incorporate biopolymers, including enzymes and monoclonal antibodies, onto the cell surface to study these puzzles. In Aim 1 of this project, we will identify or engineer a promiscuous glycosyltransferase that can transfer biopolymer-functionalized unnatural sugars from the corresponding nucleotide sugar donors to the cell-surface glycocalyx of the cells of interest. When the bioploymer is an enzyme, it will enable the proximity-dependent labeling of prey cells that interact with the cells of interest (bait cells) that harbor the enzyme, providing that the cell surface of prey cells expresses the substrates to be modified by the enzyme. Subsequently, the signal generated by the labeling can be detected and analyzed ex vivo by flow cytometry. In Aim 1d and Aim 2, we will apply this method to probe cell-cell interactions in vivo using mouse models. The focus will be the identification of interacting partners of specific immune cell types during immune activation and T-cell exhaustion (T-cell exhaustion is the process during which T cells progressively lose their functions). Finally, we will develop a method in Aim 3 to conjugate monoclonal antibodies onto immune cells to endow them with specific targeting capabilities. The migration and anti-tumor efficacy of the modified cells will be evaluated in mouse tumor models.

项目概要/摘要细胞与细胞的相互作用涉及许多生物过程，包括免疫反应、胚胎发育、神经信号传导和癌症转移。我们的理解中存在一些重大差距细胞与细胞之间的相互作用包括：特定细胞类型在不同阶段的相互作用伙伴是什么？生物过程？如何修改这些相互作用伙伴的细胞表面景观以调整功能特定细胞类型对其内源功能的干扰最小？该项目的目标是开发基于糖基转移酶的化学酶方法将生物聚合物（包括酶和单克隆抗体）整合到细胞表面以研究这些谜题。在该项目的目标 1 中，我们将鉴定或设计一种混杂的糖基转移酶，该酶可以转移生物聚合物功能化的非天然糖从相应的核苷酸糖供体到细胞表面感兴趣的细胞的糖萼。当生物聚合物是酶时，它将能够实现邻近依赖性标记与含有酶的感兴趣细胞（诱饵细胞）相互作用的猎物细胞，前提是猎物细胞的细胞表面表达要被酶修饰的底物。随后，信号可以通过流式细胞术离体检测和分析由标记产生的产物。在目标 1d 和目标 2 中，我们将应用这种方法使用小鼠模型来探测体内细胞间的相互作用。重点将是识别免疫激活和 T 细胞过程中特定免疫细胞类型的相互作用伙伴衰竭（T细胞衰竭是T细胞逐渐丧失功能的过程）。最后，我们将在目标 3 中开发一种方法，将单克隆抗体缀合到免疫细胞上，赋予它们具体的目标定位能力。修饰细胞的迁移和抗肿瘤功效将在以下方面进行评估小鼠肿瘤模型。