Core C: Subtractive Single Nucleus Seq and Spatial Multi-omics Core

项目摘要-核心C 对免疫细胞状态的有效和可重复性的检测对于我们理解免疫是至关重要的。 在健康和疾病方面的反应，如癌症和自身免疫。免疫功能是 高度依赖于共刺激信号，它们在肿瘤研究中的作用已变得明显 PD-1/PD-L1表达的浸润性淋巴细胞，以及自身免疫性疾病，尤其是 自身反应性T细胞和共刺激受体CD226及其共有的配体CD155的激活 和TIGIT在一起。实现PD-1/PD-L1和TIGIT/CD155轴调节T的机制理解 细胞和髓系细胞的功能将使这些共刺激分子的治疗用途和靶点得到改进 小路。耶鲁大学的单细胞和空间多组学核心(Core C)将为 单个细胞的转录、表观遗传和空间分布，以及ALL中丰富的免疫细胞类型 计划项目赠款中的三个项目。我们的核心已经建立了系统的管道来处理单个 细胞RNA-seq和atac-seq来自低输入样本，来自单个细胞或单个核，以及用于空间 转录学使用DBiT-seq，这是耶鲁大学新开发的单细胞空间技术，将用于 这两个实验模型和三个项目中的人体组织。该核心提供了功能和 用于准备、测序和单细胞分析以及跨物种数据整合的流水线。我们的 实验和计算资源将促进(1)小鼠模型的免疫研究， 缺乏TIGIT、PD1或两者，(2)细胞或细胞核的特征或人类组织切片 多发性硬化症和胶质母细胞瘤患者，以及(3)单细胞，综合分析产生的所有数据 在项目1-3中。总体而言，结果将是更好地理解共刺激和共抑制 人T细胞和髓系细胞炎性小体激活的调节机制及其作用 疾病和疾病模型。这可能会导致治疗自身免疫性和 癌症。

PROJECT SUMMARY – CORE C Efficient and reproducible detection of immune cell states is paramount to our understanding of the immune responses in the context of health and disease, such as cancer and autoimmunity. Immune function is highly dependent upon costimulatory signals where their role has become apparent in the study of tumor infiltrating lymphocytes with PD-1/PD-L1 expression, as well as in autoimmune disease particularly with activation of autoreactive T cells and costimulatory receptor CD226 and its ligand CD155 which is shared with TIGIT. Achieving a mechanistic understanding by which PD-1/PD-L1 and TIGIT/CD155 axes regulate T cell and myeloid cell function will enable refinement of the therapeutic use and target of these costimulatory pathways. The Single Cell and Spatial Multi-omics Core (Core C) at Yale University will provide support for novel transcriptional, epigenetic, and spatial profiling of single cells as well as enriched immune cell types across all three projects of the Program Project Grant. Our core has established systematic pipelines for processing single cell RNA-seq and ATAC-seq from low-input samples, from single cells or single nuclei, as well as for spatial transcriptomics using DBiT-seq, a single cell spatial technology newly developed at Yale which will be used in both experimental models and in human tissues across the three projects. This core provides capabilities and a pipeline for the preparation, sequencing and single cell analysis and data integration across species. Our experimental and computational resource will facilitate (1) the investigations of immunity in mouse models that are deficient for TIGIT, PD1, or both, (2) the characterization of cells or nuclei or sections human tissues from patients with multiple sclerosis and glioblastoma, and (3) the single cell, integrative analysis of all data generated in Projects 1-3. Overall, the outcome will be a better understanding of co-stimulatory and co-inhibitory mechanisms, and its function in regulating inflammasome activation in T cells and myeloid cells in human diseases and disease models. This may potentially lead to new avenues for therapy for autoimmunity and cancer.